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A multicomponent kinetic mechanism has been developed for the pyrolysis at low heating rate of carbon/phenolic thermal protection material and a wood species. An experimental campaign has been carr...

Non-edible fats are a common renewable feedstock for the biofuels production to avoid partially the use of edible feeds and fossil fuels. The aim of this work was the use of waste rendering fat to produce pyrolyzed and hydrogenated oils. The feedstock was hydrolyzed producing free fatty acids and glycerol + residues. The free fatty acids were pyrolyzed (with and without metal sulfides metal supported catalyst) or hydrotreated separately. An autoclave closed hermetically in nitrogen (pyrolysis) or hydrogen (hydrotreatment) atmosphere was used. Gaseous products were analyzed by GC‑FID/TCD. Liquid products were analyzed by Simulated Distillation (ASTM D2887) and FT-IR (attenuated total reflectance technique). For the pyrolysis, the main gaseous products were carbon dioxide, methane, ethane, and propane. For the hydrotreatment, the total amount of gases produced was much lower being the main product the carbon dioxide. For liquids, the hydrotreatment of the free fatty acids produced the respective hydrocarbons by decarboxylation reaction and the pyrolysis produced a mixture of compounds with lighter boiling ranges compared to the original free fatty acids. The use of a metal sulfide metal supported catalyst in the pyrolysis led to a higher amount of hydrogen production. but similar boiling range liquid products compared to the non-catalytic test.

Mesityl oxide is standardly used to produce methyl iso butyl ketone but it can be also used to produce other useful compounds. Three catalysts were used for the reaction of the mesityl oxide reduction. They were NiW, NiMo, and CoMo supported on phonolite modified by HCl (metals/Ph-HCl). The fresh catalysts were characterized by XRD, XRF, BET surface, Hg porosimetry, SEM, H2-TPR, NH3-TPD, CO2-TPD. The materials were directly used, previously reduced in H2 or sulfided for the mesityl oxide reduction under H2 atmosphere. The reaction was performed in an autoclave at T = 375 °C, p = 50 bar (H2), and TOS = 1.5 h. The products were analyzed by GC/MS, GC/FID-TCD, ATR. The main products were methyl isobutyl ketone, 2-methyl pentane, and 2-methyl-2-pentene. Sulfided metal catalysts were the most active in the methyl isobutyl ketone, where the NiWSx/Ph-HCl catalyst showed the highest activity. For the non-previously-activated and hydrogen activated catalysts the most active catalyst was the NiMo/Ph-HCl for the production of methyl isobutyl ketone. The catalyst CoMo/Ph-HCl activated in hydrogen was the most active for the production of 2-methyl pentane compared to the other two hydrogen-activated materials.

The oxidation of ethanol to acetaldehyde was previously investigated in a continuous flow bench scale reactor using a zirconium-oxide-supported vanadium catalyst. Products were analyzed by ...

A great emphasis is placed on searching for efficient sources to produce renewable fuels due to the actual trends in avoiding the use of fossil resources to mitigate the environment’s deterioration. Thus, the use of waste from the food or wood industry for the production of biofuels is widely researched and may contribute to sustainable general development. Rendered fat and Fischer-Tropsch waxes are high-energy materials which could be used for the pyrolysis reaction. Therefore, in this study, the effect of reaction parameters and feedstock composition on the distribution of the pyrolysis products has been examined. The experiments were carried out in a nitrogen atmosphere in a multi-shot pyrolizer instrument equipped with GC-FID. First, the influence of the temperature was examined at 785, 800, and 815 °C. The highest yield to olefins was observed at 815 °C. The effect of triglycerides and paraffins contents (0–100% with 10% ramp) in the feedstock on the product composition was investigated at 815 °C. The gas chromatography analyses revealed that the feedstock composition did not significantly affect the product composition due to the high temperature and the long linear chain structure similarities between the free fatty acids and Fischer-Tropsch waxes.

© 2021 Samperio, Guzmán-Herrador, May-Cuz, Martín, Álvarez and Llosa., Lactic acid bacteria (LAB) belonging to the genus classically known as Lactobacillus, recently split into 25 different genera, include many relevant species for the food industry. The well-known properties of lactobacilli as probiotics make them an attractive model also for vaccines and therapeutic proteins delivery in humans. However, scarce tools are available to accomplish genetic modification of these organisms, and most are only suitable for laboratory strains. Here, we test bacterial conjugation as a new tool to introduce genetic modifications into many biotechnologically relevant laboratory and wild type lactobacilli. Using mobilizable shuttle plasmids from a donor Escherichia coli carrying either RP4 or R388 conjugative systems, we were able to get transconjugants to all tested Lactocaseibacillus casei strains, including many natural isolates, and to several other genera, including Lentilactobacillus parabuchneri, for which no transformation protocol has been reported. Transconjugants were confirmed by the presence of the oriT and 16S rRNA gene sequencing. Serendipitously, we also found transconjugants into researcher-contaminant Staphylococcus epidermidis. Conjugative DNA transfer from E. coli to S. aureus was previously described, but at very low frequencies. We have purified this recipient strain and used it in standard conjugation assays, confirming that both R388 and RP4 conjugative systems mediate mobilization of plasmids into S. epidermidis. This protocol could be assayed to introduce DNA into other Gram-positive microorganisms which are resistant to transformation., Work in ML lab was supported by the grant BIO2017-87190-R from the Spanish Ministry of Science and Innovation. Work in MÁ lab was funded by the Spanish State Research Agency (AEI) and the European Regional Development Fund (FEDER) (AGL2016-78708-R, AEI/FEDER, EU). DG-H was a recipient of a predoctoral appointment from the University of Cantabria. RM-C received an Erasmus+ traineeship grant.

In this work, we report the incorporation of nickel oxide nanoparticles into a metal–organic framework (MOF) structure by a solvent-free mechanochemical strategy. In particular, the zirconium-based MOF UiO-66 was modified with different Ni loadings and characterized using complementary techniques including X-ray diffraction (XRD), N2 porosimetry and X-ray photoelectron spectroscopy (XPS). The catalytic potential of the as-prepared Ni/UiO-66 materials in the hydrogenation reaction of methyl levulinate using 2-propanol as hydrogen donor solvent has been investigated under flow conditions. Under optimized conditions, the 5%Ni/UiO-66 led to the best catalytic performance (70% yield, 100% selectivity to gamma-valerolactone), which could be attributed to the higher content of the Ni species within the MOF structure. The obtained results are promising and contribute to highlighting the great potential of MOFs in biomass upgrading processes, opening the path to the sustainable development of the chemical industry.

In this work, we present the green Ni-MOF modification by the mechanochemical process in the solvent-free route that has been prepared as hydrogenation catalyst. The zirconium-based metal-organic framework UiO-66 (MOF) at different loading of Ni, were characterized using different techniques including X-ray diffraction (XRD), N2 porosimetry, X-ray photoelectron spectroscopy (XPS), have been employed for the characterization of the structural, textural, and composition properties of the synthesized materials, respectively. The catalysts have been evaluated in the hydrogenation reaction of methyl levulinate using 2-propanol as a hydrogen donor solvent. The reaction conditions were investigated and established at 30 bar system pressure with a reaction temperature of 200 ◦C using around 0.1 g of catalyst and a flow rate of 0.2 mL/min flow rate of a 0.3 M methyl levulinate solution in 2-propanol. All catalysts employed in this work exhibited good catalytic activities under the investigated conditions, with conversion values in the 30-70% range the selectivity to Υ-valerolactone was 100% for all samples (after one hour time on stream). The highest methyl levulinate conversion and selectivity was achieved by MOF-Ni5% which can be explained by the higher dispersion content of Ni species and the highest accessibility of the Zr sites of the MOF structures even when the superficial area has been reduced.

Raman spectroscopy was used for the quantitative determination of Mo and W in Mo- and W-supported mesoporous silica (Mo/SBA-15 and W/SBA-15, respectively) and Mo-supported beta zeolite (Mo-BEA). Three Raman quantitative models were developed and optimized for the metal contents of Mo/SBA-15, W/SBA-15, and Mo/BEA. Subsequently, the models were characterized using the root mean square error of calibration (RMSEC), root mean square error of cross-validation (RMSECV), root mean square error of prediction (RMSEP), correlation coefficient, and predicted residual error sum of squares (PRESS) diagnostic function. The calibration range of the models were in the range of approximately 2&ndash, 40 wt% for the SBA-15 support and 1&ndash, 21 wt% for the BEA support because the BEA support presented lower Mo absorption than the SBA-15 support. The RMSEC, RMSECV, and RMSEP values were below 1.80% for all developed models. The highest and lowest correlation coefficients corresponded to the W/SBA-15 (0.9984) and Mo/BEA (0.9777) models, respectively. The change in catalyst support affected the mentioned chemometric parameters (Mo/SBA-15 vs. Mo/BEA). Subsequently, Raman spectroscopy combined with the temperature control stage was used to study the calcination of Mo/BEA, Mo/SBA-15, and W/SBA-15 using three-dimensional diagrams, in which the changes in catalyst structure were analyzed as functions of the temperature and time. Raman spectroscopy was determined to be a suitable analytical tool for the quantitative analysis of the metal contents of the catalyst and optimization of the calcination process. Therefore, Raman spectroscopy can be used during catalyst manufacturing.

The use of renewable local raw materials to produce fuels is an important step toward optimal environmentally friendly energy consumption. In addition, the use of these sources together with fossil fuels paves the way to an easier transition from fossil to renewable fuels. The use of simple organic acids as hydrogen donors is another alternative way to produce fuel. The present work reports the use of oxalic acid as a hydrogen donor for the catalytic hydrodesulfurization of atmospheric gas oil and the deoxygenation of rapeseed oil at 350 &deg, C. For this process, one commercial NiW/SiO2&ndash, Al2O3 solid and two NiW/modified phonolite catalysts were used, namely Ni (5%) W (10%)/phonolite treated with HCl, and Ni (5%) W (10%)/phonolite treated with oxalic acid. The fresh phonolite catalysts were characterized by Hg porosimetry and N2 physisorption, ammonia temperature programmed desorption (NH3-TPD), X-ray diffraction (XRD), and X-ray fluorescence (XRF). The sulfided metal phonolite catalysts were characterized by XRD and XRF. Hydrodesulfurization led to a decrease in sulfur content from 1 to 0.5 wt% for the phonolite catalysts and to 0.8 wt% when the commercial catalyst was used. Deoxygenation led to the production of 15 and 65 wt% paraffin for phonolite and commercial solids, respectively. The results demonstrate the potential of using oxalic acid as a hydrogen donor in hydrotreating reactions.

Highlights • Alphavirus self-amplifiying RNA (saRNA) induces more potent immune responses than conventional mRNA. • saRNA delivery can be enhanced by electroporation or conjugation with cationic lipid or polymers. • saRNA vaccines induce protective responses against human pathogens in preclinical models. • saRNA replication mediates innate immune signals that contribute to the strength of immune responses. • saRNA vaccines could be generated in a quick way to face emergent pathogens like SARS-CoV-2., DNA or mRNA vaccines have potential advantages over conventional vaccines since they are easier to manufacture and have higher safety profiles. In particular, self-amplifying RNA (saRNA) derived from alphavirus expression vectors has shown to be very efficient to induce humoral and cellular responses against many antigens in preclinical models, being superior to non-replicating mRNA and DNA. This is mainly due to the fact that saRNA can provide very high expression levels and simultaneously induces strong innate responses, potentiating immunity. saRNA can be administered as viral particles or DNA, but direct delivery as RNA represents a safer and more simple approach. Although saRNA can be delivered as naked RNA, in vivo transfection can be enhanced by electroporation or by complexing it with cationic lipids or polymers. Alphavirus saRNA could have broad application to vaccinate against human pathogens, including emerging ones like SARS-CoV-2, for which clinical trials have been recently initiated.

Direct coal liquefaction (DCL) seems to be a suitable way to convert low rank coals into liquid fuels, especially when problematic wastes are used together with this feedstock. It is also a solution to become energy independent for many countries which have significant coal resources and limited access to crude oil. The aim of this research was to investigate DCL process by the co-processing of brown coal with model-, petroleum- and waste-derived solvents. The brown coal with and without W, Fe and Mo metals supported on its surface was tested. Thirteen DCL tests with added hydrogen pressure were carried out in the autoclave. These tests were classified in two groups. First group covered reactions using model and petroleum-derived solvents (tetralin, 1-methylnaphthalene, light cycle oil (LCO), hydrotreated LCO, C9+ fraction and decalin) with non-impregnated brown coal. Then, LCO was chosen as solvent for carrying out three tests using the metals supported on coal. Finally, waste tires pyrolysis oil was used as a waste-derived solvent for other three tests with molybdenum supported on coal. For tests using LCO, the total amount of direct liquid and n-heptane soluble products was 20 wt% higher using metal covered brown coal in comparison to unmodified one. The test with the brown coal impregnated by 1% of molybdenum resulted in the best efficiency, thus this type of coal was chosen as a catalytic feedstock for the tests with waste tires pyrolysis oil. info:eu-repo/semantics/publishedVersion

Thermal Protection Materials (TPM) such as carbon/phenolic composites are used to protect spacecraft structures from extreme conditions. This protection is, in part, achieved by the decomposition via pyrolysis of the phenolic resin. Finite rate chemistry models are however still unable to predict the chemical production rates and composition of the pyrolysis products accurately. This is mostly due to the scarcity of experimental data for model validation. In this work, the decomposition of a phenolic material representative of thermal protection material is studied in a unique micro-pyrolysis unit for the temperature range 300-800 °C. This unit is equipped with highly sensitive detectors allowing us to identify and quantify products in a broad range of molecular weights up to 240 g mol−1. More than 50 different products of the pyrolysis of phenolic resin have been quantified with a mass balance closure greater than 80%. The major compound groups found are permanent gases, phenols as well as larger molecules such as diphenols and naphthalenes. In addition, the char yield obtained at the fast heating rates employed in our apparatus was found ∼ 5 % -points lower compared to traditional thermogravimetry.

Two VOx/SBA-15 catalysts and three VOx/SiO2-fumed silica, with 5, 10 and 1, 5, 10 %wt. vanadium content respectively, were tested in a stainless steel continuous flow reactor for the partial oxidation of ethanol. The catalysts were tested at 150 – 300 °C. Products were analyzed by GC-FID, GC-OFID and GC-MS. The aim was exploring the problematics which could be found when more industrial close conditions are used. The total conversion of ethanol and selectivity to acetaldehyde were different than the expected ones. For VOx/SiO2-fumed silica, the total conversion was higher with a lower selectivity to acetaldehyde compared to VOx/SBA-15 catalysts.

The potential of co-producing two different biofuels from a lignocellulosic substrate (wheat straw), according to a biorefinery concept, has been investigated. For such a purpose, simultaneous saccharification and fermentation (SSF) from the hemicellulosic and cellulosic fractions was performed for maximizing bioethanol production. The non-washed water-insoluble solid (WIS) fraction from the pretreated wheat straw totally inhibited the production of ethanol by Kluyveromyces marxianus independently of the inoculum size. In contrast, when using washed-WIS, higher ethanol productivities at 24 h of SSF were attained when increasing the inoculum size from 1 g/L to 3 g/L. The residual lignin from the bioethanol process was transformed by fast-pyrolysis into bio-oil that can be further converted into other biofuels or biochemicals. Thermal fast-pyrolysis of the residual lignin fraction produced 31.9 wt% of bio-oil ∗ (water free basis) mostly composed by oxygenated aromatics coming from the lignin monomers (guaiacol-, syringol- and phenol-derived compounds). On the other hand, catalytic fast-pyrolysis of the residual lignin fraction over HZSM-5 zeolite was used as preferentially promoted decarbonylation and cracking of the primary vapours. Coupling both processes significantly enhanced the production of liquid products from lignocellulose, improving the efficiency in the use of the raw material. In this way, compared to a simple process of bioethanol production, this approach allowed to increase the mass and the chemical energy yields 1.9 and 1.7-fold, respectively.

A novel model for the pyrolysis of the Phenolic Impregnated Carbon Ablator (PICA) at high heating rate is developed and calibrated based on high fidelity thermal decomposition experiments. The calibration relies on accurate quantification of pyrolysis gases obtained from mass spectroscopy analysis during thermal decomposition at fast heating rates simulating flight conditions. Model calibration is achieved by coupling the Porous material Analysis Toolbox based on OpenFOAM (PATO) with an optimization software (Dakota). A multi-objective genetic algorithm is used to fit the experimental data by optimizing the model parameters for an element and a species-based formulation. The new model captures both the material mass loss and the gaseous species produced during pyrolysis., SCOPUS: ar.j, info:eu-repo/semantics/published

This work was supported by grants from Ministerio de Ciencia e Innovacion, Spain, and the ERDF (RTI2018-093503-B-100), the Muscular Dystrophy Association (MDA-602322). C.M.Q. is supported by the Department of Defense (DOD) grant PR190511. A.H.-G. and P.G.-G. are `FPU fellows' from the Ministerio de Universidades, Spain. S.L.-H. is supported by the "becas de colaboracion" from the Ministerio de Universidades, Spain. E.B.-C. is supported by the Consejeria de Salud, Junta de Andalucia, Spain., We thank Stacy Kelly Aguirre for the English editing. Figures created with BioRender.com., Coenzyme Q10 (CoQ(10)) is classically viewed as an important endogenous antioxidant and key component of the mitochondrial respiratory chain. For this second function, CoQ molecules seem to be dynamically segmented in a pool attached and engulfed by the super-complexes I + III, and a free pool available for complex II or any other mitochondrial enzyme that uses CoQ as a cofactor. This CoQ-free pool is, therefore, used by enzymes that link the mitochondrial respiratory chain to other pathways, such as the pyrimidine de novo biosynthesis, fatty acid beta-oxidation and amino acid catabolism, glycine metabolism, proline, glyoxylate and arginine metabolism, and sulfide oxidation metabolism. Some of these mitochondrial pathways are also connected to metabolic pathways in other compartments of the cell and, consequently, CoQ could indirectly modulate metabolic pathways located outside the mitochondria. Thus, we review the most relevant findings in all these metabolic functions of CoQ and their relations with the pathomechanisms of some metabolic diseases, highlighting some future perspectives and potential therapeutic implications., Spanish Government, European Commission RTI2018-093503-B-100, Muscular Dystrophy Association MDA-602322, United States Department of Defense PR190511, Ministerio de Universidades, Spain, Junta de Andalucia

The oxidative cleavage of β-carotene in the Mucorales produces three fragments of 18, 15, and 7 carbons, respective heads of three families of apocarotenoids: the methylhexanoids, the trisporoids, and the cyclofarnesoids (named after their 1,6-cyclofarnesane skeleton). The apocarotenoids are easily recognized because they are absent in white mutants unable to produce β-carotene. In cultures of Phycomyces blakesleeanus we detected thirty-two apocarotenoids by LC, UV absorbance, and MS. With additional IR and NMR we identified two methylhexanoids and the eight most abundant cyclofarnesoids. Four of them were previously-unknown natural compounds, including 4-dihydrocyclofarnesine S, the most abundant cyclofarnesoid in young cultures. We arranged the apocarotenoids of the Mucorales in a scheme that helps classifying and naming them and suggests possible metabolites and biosynthetic pathways. We propose specific biosynthetic pathways for cyclofarnesoids and methylhexanoids based on structural comparisons, the time course of appearance of individual compounds, and the bioconversion of β-apo-12-carotenol, an early precursor, to three more oxygenated cyclofarnesoids by the white mutants. Some of the reactions occur spontaneously in the increasingly acidic culture media. Mating increased the contents of methylhexanoids and cyclofarnesoids by ca. threefold in young cultures and ca. twelvefold in old ones (five days); cyclofarnesine S, the most abundant cyclofarnesoid in old cultures, increased over one hundredfold. We found no differences between the sexes and no activity as sexual pheromones, but we suggest that methylhexanoids and cyclofarnesoids could mediate species-specific physiology and behavior.

Oxalic acid was employed for the catalytic transfer hydrogenation of used vegetable cooking oil. Hydrogen production was achieved upon the decomposition of oxalic acid using a commercial industrially sulfided NiW/SiO2-Al2O3 catalyst. Products were analysed by attenuated infrared spectroscopy, simulated distillation and gas chromatography-flame ionisation detection/thermal conductivity detection for the produced gases. No hydrogen gas was added for these tests, which were performed in a nitrogen atmosphere at an initial pressure of 0 bar in an autoclave. Unprecedented results were obtained, with a high yield of deoxygenated hydrotreated vegetable oil hydrocarbons being achieved while avoiding the use of formic acid, which is both toxic and irritant.

Due to the high costs and low selectivity associated with the production of propylene, new routes for its synthesis are being sought. Dehydration has been widely investigated in this field, but, thus far, no study has produced efficient results for isopropanol. Vanadium-zirconia catalysts have been shown to be effective for the dehydration of ethanol. Therefore, we investigated the activity of such catalysts in the dehydration of isopropanol. The catalysts were synthetized on a SBA-15 base, supplemented with zirconia or combined zirconia and vanadium. Tests were conducted in a continuous flow reactor at 150&ndash, 300 &deg, C. Samples were analyzed using a gas chromatograph. The most active catalyst showed 96% conversion with 100% selectivity to propylene. XRD, SEM and Raman spectroscopy analyses revealed that as the vanadium content increases, the pore size of the catalyst decreases and both isopropanol conversion and propylene selectivity are reduced. Thus, without the addition of vanadium, the Zr-SBA-15 catalyst appears to be suitable for the dehydration of isopropanol to propylene.

Coal and waste direct liquefaction was carried out to decrease the hydrocarbons production negative bearing to environment. The wastes studied were glycerol, polyethylene waste and waste tyres pyrolysis oil. Glycerol and waste tyres pyrolysis oil were blended with tetralin and used as solvent for coal direct liquefaction (T = 350–450 °C; N2; waste:tetralin at different weight ratios). Glycerol may be considered as a waste from the production of fatty acid methyl esters (FAME). Waste tyres pyrolysis oil showed to be a good hydrogen donor solvent. Solid polyethylene wastes were mixed with coal and used in coal co-liquefaction tests, which showed to be a good option to produce liquid hydrocarbons for fuels or raw materials for chemical industry. Although liquids yield using waste tyres pyrolysis oil or glycerol were high, respectively 62% wt. and 51% wt., the use of polyethylene allowed increasing them to 80% wt. Different experimental conditions were studied, namely: experimental temperature, run time, solvent type and composition and hydrogen initial pressure. The effect of commercial catalysts on co-liquefaction was also studied and the results obtained were compared with those obtained with coal impregnated with Fe or Mo (1% wt./coal). Response surface methodology (RSM) was used to predict the influence of the main experimental conditions studied on product yields. This methodology resulted to be a good one to predict coal liquefaction and co-liquefaction products yields, as predicted values were similar to experimental data. In addition, RSM approach led to the selection of tetralin as solvent and to the use of coal impregnated with 1% of Mo, as liquids yield increased up to 96% wt.

Carbon/phenolic ablators are successfully used as thermal protection material for spacecraft. Nevertheless, their complex thermal degradation is not yet fully understood, and current pyrolysis models do not reproduce important features of available experimental results. Accurate and robust thermal degradation models are required to optimize design margin policy. We investigate whether the competitive kinetic schemes commonly used to model biomass pyrolysis are appropriate to describe the thermal degradation of carbon/phenolic composites. In this paper, we apply competitive pyrolysis mechanisms for the thermal degradation of the carbon/phenolic ablator PICA. Model parameters are then calibrated using a robust two-step methodology: first deterministic optimization is used to obtain the best estimation of the calibration parameters based on the experimental data, then a stochastic Bayesian inference is performed to explore plausible set of solutions taking into account the experimental uncertainties. The proposed calibrated model provides an accurate description of the pyrolysis process at different heating rates. The model shows great flexibility and robustness at a similar computational cost as the traditional devolatilization models. This opens the possibility for more complex mechanisms when more experimental data becomes available.

Conjugative relaxases are well-characterized proteins responsible for the site- and strand-specific endonucleolytic cleavage and strand transfer reactions taking place at the start and end of the conjugative DNA transfer process. Most of the relaxases characterized biochemically and structurally belong to the HUH family of endonucleases. However, an increasing number of new families of relaxases are revealing a variety of protein folds and catalytic alternatives to accomplish conjugative DNA processing. Relaxases show high specificity for their cognate target DNA sequences, but several recent reports underscore the importance of their activity on secondary targets, leading to widespread mobilization of plasmids containing an oriT-like sequence. Some relaxases perform other functions associated with their nicking and strand transfer ability, such as catalyzing site-specific recombination or initiation of plasmid replication. They perform these roles in the absence of conjugation, and the validation of these functions in several systems strongly suggest that they are not mere artifactual laboratory observations. Other unexpected roles recently assigned to relaxases include controlling plasmid copy number and promoting retrotransposition. Their capacity to mediate promiscuous mobilization and genetic reorganizations can be exploited for a number of imaginative biotechnological applications. Overall, there is increasing evidence that conjugative relaxases are not only key enzymes for horizontal gene transfer, but may have been adapted to perform other roles which contribute to prokaryotic genetic plasticity. Relaxed target specificity may be key to this versatility., Work in our lab is supported by grants BIO2017-87190-R from the MINECO (Spanish Ministry of Economy and Innovation), and IDEAS211LLOS from the AECC (Spanish Association Against Cancer) to ML. DLG-H is a recipient of a predoctoral appointment from the University of Cantabria.

Background The current standard treatment for patients with chronic hepatitis C virus (HCV) infection is a combination of direct-acting antiviral agents(DAAs). Still, a relatively small amount of patients fail DAAs and there is not enough data to establish treatment recommendations for these cases. Purpose The aim is to describe the efficacy of a second round of DAAs treatment for HCV infection after unsuccessful therapy with DAAs agents. Material and methods An observational retrospective descriptive study was performed. The patients included in this study were diagnosed with HCV, underwent a first treatment with DAAs from September 2014 to March 2017, but did achieve sustained virological response 12 weeks after treatment (SVR12). The variables genotype, liver fibrosis (METAVIR score), first treatment with DAAs, SVR12 and second round of treatment, were extracted from electronic health records. Results A total of 352 patients received treatment with DAAs and of these, 14 did not achieve SVR12. 4/14 did not receive new treatment, 3/14 received new treatment for 12 weeks and 7/14 for 24 weeks. The DAAs used were: simeprevir (SMV), sofosbuvir (SOF), ledipasvir (LED), ribavirina (RBV), dasabuvir (DSB), ombitasvir/paritaprevir/ritonavir (OMB/PTV/r), peginterferon (PEG), daclatasvir (DCV) and velpatasvir (VEL). The results by genotype were: genotype 1 (9/14): three patients had genotype 1a and six patients had genotype 1b. Regarding liver fibrosis diagnosis, five patients were F4, one patient was F3, two were patients F3 and one patient without Fibroscan ® . Regarding treatment: 3/9 initially received SMV +SOF, and after SVR12 failure, 2/3 received LED/SOF +RBV and 1/3 received OMB/PTV/r+RBV. 2/9 received a first treatment with SMV +c +RBV and then LED/SOF +RBV. 1/9 received a first treatment with SOF +RBV and then DCV +SOF. 3/9 received initially LED/SOF and after failure, 1/3 received SMV +SOF + RBV, 1/3 received DCV +SMV +SOF and 1/3 did not receive new treatment. Genotype 2 (1/14): one F4 patient received SOF+RBV and then DCV+SOF+ RBV. Genotype 3 (3/14): two patients were F2 and 1 was F4. Patients first received DCV+SOF and then 2/3 received SOF/VEL+RBV, and 1/3 did not receive new treatment (currently under evaluation). Genotype 4 (1/14): one F3 patient received LED/SOF and then SMV+SOF+ RBV. Efficacy data were available for 10/14 patients and all of them achieved SVR12 with the second round of DAAs treatment. Conclusion In patients who failed to achieve a SVR12 with first treatment, the second round of DAAs treatment was very successful. However, more data are necessary to establish strong recommendations. No conflict of interest

Several preparations were obtained from the aerial parts of predomesticated Lavandula luisieri, including the essential oil and ethanolic, hexane, and ethyl acetate extractives. Additionally, pilot plant vapor pressure extraction was carried out at a Several preparations were obtained from the aerial parts of predomesticated Lavandula luisieri, including the essential oil and ethanolic, hexane, and ethyl acetate extractives. Additionally, pilot plant vapor pressure extraction was carried out at a pressure range of 0.5–1.0 bar to give a vapor pressure oil and an aqueous residue. A chemical study of the hexane extract led to the isolation of six necrodane derivatives (1, 2, and 4–7), with four of these (1, 2, 5, and 7) being new, as well as camphor, a cadinane sesquiterpene (9), tormentic acid, and ursolic acid. The EtOAc and EtOH extracts contained a mixture of phenolic compounds with rosmarinic acid being the major component. Workup of the aqueous residue resulted in the isolation of the necrodane 3 and (1R*,2S*,4R*)-p-menth-5-ene-1,2,8-triol (8), both new natural compounds. The structures of the new compounds were established based on their spectroscopic data. The phytotoxic and nematicidal activities of these compounds were evaluated., This work has been partially supported by grants CTQ2012- 38219-C03-01 (Spain), Junta de Andalucia (Excellence Project ́ P08-FQM-3596) Spain, and JAE-CSIC (predoctoral fellowship to L.F.J.).

Poly(ADP-ribosyl)ation (PAR) has been implicated in various aspects of the cellular response to DNA damage and genome stability. Although 17 human poly(ADP-ribose) polymerase (PARP) genes have been identified, a single poly(ADP-ribosyl) glycohydrolase (PARG) mediates PAR degradation. Here we investigated the role of PARG in the replication of human chromosomes. We show that PARG depletion affects cell proliferation and DNA synthesis, leading to replication-coupled H2AX phosphorylation. Furthermore, PARG depletion or inhibition per se slows down individual replication forks similarly to mild chemotherapeutic treatment. Electron microscopic analysis of replication intermediates reveals marked accumulation of reversed forks and single-stranded DNA (ssDNA) gaps in unperturbed PARG-defective cells. Intriguingly, while we found no physical evidence for chromosomal breakage, PARG-defective cells displayed both ataxia-telangiectasia-mutated (ATM) and ataxia-Rad3-related (ATR) activation, as well as chromatin recruitment of standard double-strand-break-repair factors, such as 53BP1 and RAD51. Overall, these data prove PAR degradation to be essential to promote resumption of replication at endogenous and exogenous lesions, preventing idle recruitment of repair factors to remodeled replication forks. Furthermore, they suggest that fork remodeling and restarting are surprisingly frequent in unperturbed cells and provide a molecular rationale to explore PARG inhibition in cancer chemotherapy.

A sample of phonolite was treated by cold plasma and hydrochloric acid diluted in water to study the change of its structure and acid properties. The phonolite and treated samples were analysed by XRD, elemental analysis XRF, specific surface area BET, TPD-NH 3 and FT-IR spectroscopy. They were also tested in the adsorption of Ca, K, Mg, P and Na impurities present in waste cooking oil. Plasma treated sample presented almost the same structure with some surface differences respect to the original phonolite. However, acid treated sample presented bigger total surface compared to the other samples, different structure, composition and acid properties.

BACKGROUND: The emergence of drug resistance in Plasmodium falciparum has been a major contributor to the global burden of malaria. Drug resistance complicates treatment, and it is one of the most important problems in malaria control. This study assessed the level of mutations in P. falciparum genes, pfdhfr, pfdhps, pfmdr1, and pfcrt, related to resistance to different anti-malarial drugs, in the Continental Region of Equatorial Guinea, after 8 years of implementing artesunate combination therapies as the first-line treatment. RESULTS: A triple mutant of pfdhfr (51I/59R/108N), which conferred resistance to sulfadoxine/pyrimethamine (SP), was found in 78% of samples from rural settings; its frequency was significantly different between urban and rural settings (p = 0.007). The 164L mutation was detected for the first time in this area, in rural settings (1.4%). We also identified three classes of previously described mutants and their frequencies: the partially resistant (pfdhfr 51I/59R/108N + pfdhps 437G), found at 54% (95% CI 47.75-60.25); the fully resistant (pfdhfr 51I/59R/108N + pfdhps 437G/540E), found at 28% (95% CI 7.07-14.93); and the super resistant (pfdhfr 51I/59R/108N + pfdhps 437G/540E/581G), found at 6% (95% CI 0.48-4.32). A double mutation in pfmdr1 (86Y + 1246Y) was detected at 2% (95% CI 0.24-3.76) frequency, distributed in both urban and rural samples. A combination of single mutations in the pfmdr1 and pfcrt genes (86Y + 76T), which was related to resistance to chloroquine and amodiaquine, was detected in 22% (95% CI 16.8-27.2) of samples from the area. CONCLUSIONS: The high level of mutations detected in P. falciparum genes related to SP resistance could be linked to the unsuccessful withdrawal of SP treatment in this area. Drug resistance can reduce the efficacy of intermittent prophylactic treatment with SP for children under 5 years old and for pregnant women. Although a high number of mutations was detected, the efficacy of the first-line treatment, artemisinin/amodiaquine, was not affected. To avoid increases in the numbers, occurrence, and spread of mutations, and to protect the population, the Ministry of Health should ensure that health centres and hospitals are supplied with appropriate first-line treatments for malaria. We would like to thank the National Malaria Control Programme and Ministry of Health and Ministry of Health and Social Welfare of Equatorial Guinea for their assistance. We would also like to thank the Network of Tropical Diseases Research Centres (Red de Investigación Cooperativa en Enfermedades Tropicales/ RICET-: RD12/0018/0001). This work has been done under the project: PI14CIII/00064-TRPY 1282/15 and was funded by the Institute of Health Carlos III. Sí

We explore the potential of bacterial secretion systems as tools for genomic modification of human cells. We previously showed that foreign DNA can be introduced into human cells through the Type IV A secretion system of the human pathogen Bartonella henselae. Moreover, the DNA is delivered covalently attached to the conjugative relaxase TrwC, which promotes its integration into the recipient genome. In this work, we report that this tool can be adapted to other target cells by using different relaxases and secretion systems. The promiscuous relaxase MobA from plasmid RSF1010 can be used to deliver DNA into human cells with higher efficiency than TrwC. MobA also promotes DNA integration, albeit at lower rates than TrwC. Notably, we report that DNA transfer to human cells can also take place through the Type IV secretion system of two intracellular human pathogens, Legionella pneumophila and Coxiella burnetii, which code for a distantly related Dot/Icm Type IV B secretion system. This suggests that DNA transfer could be an intrinsic ability of this family of secretion systems, expanding the range of target human cells. Further analysis of the DNA transfer process showed that recruitment of MobA by Dot/Icm was dependent on the IcmSW chaperone, which may explain the higher DNA transfer rates obtained. Finally, we observed that the presence of MobA negatively affected the intracellular replication of C. burnetii, suggesting an interference with Dot/Icm translocation of virulence factors., This work was supported by grant BIO2013-46414-P from the Spanish Ministry of Economy and Competitiveness to ML, and NIH grants AI041699 and AI114760 to CRR. DLG was supported by a predoctoral fellowship from the University of Cantabria (Spain). SS was supported by an Advanced Postdoc Mobility fellowship from the Swiss National Science Foundation (SNSF).

Conjugative transfer of plasmid R388 requires the coupling protein TrwB for protein and DNA transport, but their molecular role in transport has not been deciphered. We investigated the role of residues protruding into the central channel of the TrwB hexamer by a mutational analysis. Mutations affecting lysine residues K275, K398, and K421, and residue S441, all facing the internal channel, affected transport of both DNA and the relaxase protein in vivo. The ATPase activity of the purified soluble variants was affected significantly in the presence of accessory protein TrwA or DNA, correlating with their behaviour in vivo. Alteration of residues located at the cytoplasmic or the inner membrane interface resulted in lower activity in vivo and in vitro, while variants affecting residues in the central region of the channel showed increased DNA and protein transfer efficiency and higher ATPase activity, especially in the absence of TrwA. In fact, these variants could catalyze DNA transfer in the absence of TrwA under conditions in which the wild-type system was transfer deficient. Our results suggest that protein and DNA molecules have the same molecular requirements for translocation by Type IV secretion systems, with residues at both ends of the TrwB channel controlling the opening–closing mechanism, while residues embedded in the channel would set the pace for substrate translocation (both protein and DNA) in concert with TrwA., This work was supported by Grants BIO2013-46414-P, BFU2016-78521-R, and BFU2014-55534-C2-1-P from the Spanish Ministry of Economy and Competitiveness to ML, EC, and FdlC, respectively. GL acknowledges funding from NIH Grants GM030518, S10OD012351, and S10OD021764. DL was a recipient of a predoctoral fellowship from CSIC (JAE-PRE). DLG-H and IM were recipients of predoctoral fellowships from the University of Cantabria.

The heterocyclic C15 apocarotenoid 1 was isolated from mated cultures of the strains F986 (+) and F921 (−) of Blakeslea trispora. This new compound formed during sexual interaction is a minor constituent of the culture media and its structure was elucidated by spectroscopic data, including 2D-NMR. A plausible biosynthetic pathway involving a double degradation of β-carotene, followed by several oxidations of the resulting monocyclofarnesane C15 fragment is proposed., This research was financed by Junta de Andalucía (Grants FQM 340, CVI 910, and P08-CVI-03901) and the Spanish Government (Grant CTQ 2010-16818, subprogram BQ).

This article presents for the first time the joint use of ionic liquid-based single drop microextraction (IL-SDME) and ion mobility spectrometry (IMS) for the determination of 2,4,6-trichloroanisol (2,4,6-TCA) in water and wine samples. An imidazolium-based IL was used as extractant taking into account both, its affinity for the analyte and its negligible response in the detector. Water samples were directly analyzed by IL-SDME while wines required a previous solid phase extraction step to remove the interference of ethanol in the ion mobility spectra. All the parameters affecting the extraction were optimized in order to achieve the highest sensitivity, taking into account the lower concentrations reported for this compound in the matrices selected. Moreover, the use of ion mobility working under the negative ionization mode confers an adequate selectivity. The limits of detection and quantification were 0.2 and 0.66 ng L(-1), respectively. The precision of the method calculated at 10 ng L(-1) was 1.4% (repeatability, n=5) and 2.2% (reproducibility, n=5, 3 days). The analysis of water and wine samples reported the presence of 2,4,6-TCA in wine samples commercialized in bottles with cork stoppers.

Advances and changes in practical aspects of ion-mobility spectrometry (IMS) have led to its widespread use for applications of environmental concern due to its unique characteristics, which include portability, ruggedness, relatively low acquisition costs and speed of analysis. However, limitations regarding the complexity of environmental samples and strict requirements on limits of detection have to be overcome. This article critically reviews existing environmental applications using IMS for the determination of different families of compounds. We also consider the analytical tools developed to solve the limitations regarding selectivity and sensitivity, including those approaches that have led to advances in the instrumentation of IMS and its combination with other techniques for extraction and pre-concentration of analytes, pre-separation of analytes and its coupling to other detection systems. Finally, we discuss current trends that facilitate the deployment of IMS for on-site or in-field analysis.

The roles that ionic liquids (ILs) can play as extractants, intermediate solvents, mediators and desorption solvents in liquid-phase and solid-phase microextraction (SPME) provide the topic of this review. We emphasize the added value of using ILs in single-drop microextraction, hollow-fiber-based liquid-phase microextraction, dispersive liquid-liquid microextraction, and in-fiber and in-tube SPME, the most relevant being greater simplicity, greater selectivity, greater extractability and lower consumption of organic solvents. We also give systematic consideration to the requirements of detection as the next step. Further, we outline the unique properties of ILs that are exploited in each extraction procedure. The growing importance of microextraction techniques in sample preparation justifies this approach.

The combination of ionic liquid-based headspace single drop microextraction (IL-HS-SDME) and room-temperature gas chromatography/ion mobility spectrometry (RTGC-IMS) is presented for the first time using the direct determination of trihalomethanes in waters as model analytical problem. The ionic liquid allows the transference of the analytes from the sample to the analytical system, at the same time that it provides an increase of the sensitivity and selectivity of the determination. An injection unit has been designed to permit the efficient volatilization of the analytes at room temperature and to avoid the entering of IL in the system. The direct combination allows the determination of the halocompounds in a rapid and simple way taking advance of their characteristic IMS spectra. The limits of the detection range between 0.1 ng mL −1 (bromoform) and 0.9 ng mL −1 (chloroform), the reproducibility of the system being better than 7.1% (RSD). The proposed coupling opens up a new horizon in IMS-based applications.

An efficient procedure for the isolation of the sesquiterpene ilicic acid (3) on a multigram scale of extracts obtained from aerial parts of Inula viscosa (Asteraceae) was developed. Acid 3 is an appropriate starting material for short, enantiospecific syntheses of β-eudesmol (1) and α-eudesmol (2), natural products featuring significant antiangiogenic and anti-Alzheimer properties. Synthesis of 1 was achieved in six steps and the synthesis of 2 in seven, producing overall yields of 52 and 41 %, respectively.(© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)