The use of functionalized carbon xerogels in cells growth

In the present study carbon xerogels are used for the first time to study the fibroblast cell growth. For that, carbon xerogel microspheres are synthesized and thereafter functionalized with carbon nanofibers followed by the 1,3-dipolar cycloaddition of azomethine ylides (the so called \u201cPrato reaction\u201d) or the addition of aryl diazonium salts (the so called \u201cTour reaction\u201d) to improve its wettability. The presence of nanofibers produces a huge improvement of the functionalization degree (59 versus 372 \u3bcmol/g for pristine carbon spheres and carbon spheres with 30% of carbon nanofibers, respectively) in spite of the blockage of the carbon spheres porosity caused after the nanofibers growth. This improvement was explained on the base of the increase of the number of probable active sites for the addition reactions (C[dbnd]C bonds) and the accessibility to these active sites (accessible surface area) by the presence of nanofibers. These high functionalization degrees reflect a promising potential of these materials in biomedical applications. Toxicity results obtained using a fibroblast cell line showed that samples are biocompatible for this kind of cells and that the presence of carbon fibers on the surface of the spheres increases the cells proliferation in a high extend reaching in some case values around 150% regarding the control. This study evidences that carbon aerogels could be interesting materials in biological applications, an unexplored field for this type of materials, being biocompatible, favouring the proliferation of cells and achieving high functionalization degrees

Biogas upgrading by selective adsorption onto CO2 activated carbon from wood pellets

An activated carbon from commercial pine wood pellets was obtained by means of physical activation with carbon dioxide. Carbonized sample was also prepared in order to compare both samples. Carbonization lead to a material with very low micropore volume, which was increased when physical activation was performed (W-0(N-2) from 0.00 to 0.22 cm(3)g (1) and W-0(CO2) from 0.21 to 0.31 cm(3) g (1)). After characterizing their porous texture, samples were tested as CO2 selective adsorbent in a mixture of CH4 and CO2; at first high pressure isotherms of both compounds were obtained at different temperatures to calculate the adsorbed amount of each compound at given pressure and temperature (for PINPEL20: CO2, q(sat) = 6.66 mol kg (1) and Delta H-ads = 28.4 kJ mol (1); CH4, q(sat) = 3.36 mol kg (1) and Delta H-ads = 19.0 kJ mol (1)). Variations on isosteric heat of adsorption with uptake were also analysed (it was increased with uptake in the case of CO2 and the opposite tendency was found for CH4). Selectivity resulted larger for the activated sample (2.9 versus 1.9 at 1 bar and 30 degrees C), so this one was chosen to perform dynamic adsorption experiments. Breakthrough curves demonstrated the outstanding performance of sample as CO2 selective adsorbent in both single component experiments (Breakthrough time 995 s for CH4 and 1545 s for CO2) and competitive adsorption (950 s for CH4 and 1373 s for CO2). Activated sample showed an excellent behaviour as CO2 selective adsorbent for biogas upgrading

Characterization of the responses to saline stress in the symbiotic green microalga Trebouxia sp. TR9

[EN] Main conclusion. For the first time we provide a study on the physiological, ultrastructural and molecular effects of salt stress on a terrestrial symbiotic green microalga, Trebouxia sp. TR9. Although tolerance to saline conditions has been thoroughly studied in plants and, to an extent, free-living microalgae, scientific data regarding salt stress on symbiotic lichen microalgae is scarce to non-existent. Since lichen phycobionts are capable of enduring harsh, restrictive and rapidly changing environments, it is interesting to study the metabolic machinery operating under these extreme conditions. We aim to determine the effects of prolonged exposure to high salt concentrations on the symbiotic phycobiont Trebouxia sp. TR9, isolated from the lichen Ramalina farinacea. Our results suggest that, when this alga is confronted with extreme saline conditions, the cellular structures are affected to an extent, with limited chlorophyll content loss and photosynthetic activity remaining after 72h of exposure to 5M NaCl. Furthermore, this organism displays a rather different molecular response compared to land plants and free-living halophile microalgae, with no noticeable increase in ABA levels and ABA-related gene expression until the external NaCl concentration is raised to 3M NaCl. Despite this, the ABA transduction pathway seems functional, since the ABA-related genes tested are responsive to exogenous ABA. These observations could suggest that this symbiotic green alga may have developed alternative molecular pathways to cope with highly saline environments.Supported by the Ministerio de Economía y Competitividad (MINECO, Spain) and FEDER (CGL2016-79158-P), and the PROMETEO Excellence in Research Program (Generalitat Valenciana, Spain) (PROMETEO/2017/039). Funding for Ernesto Hinojosa-Vidal was also provided by MINECO (BES-2013-065511).Hinojosa-Vidal, E.; Marco, F.; Martínez-Alberola, F.; Escaray, F.; García-Breijo, F.; Reig-Armiñana, J.; Carrasco, P.... (2018). Characterization of the responses to saline stress in the symbiotic green microalga Trebouxia sp. TR9. Planta. 248(6):1473-1486. https://doi.org/10.1007/s00425-018-2993-8S147314862486Álvarez R, del Hoyo A, Díaz-Rodríguez C et al (2015) Lichen rehydration in heavy metal-polluted environments: Pb modulates the oxidative response of both Ramalina farinacea thalli and its isolated microalgae. Microb Ecol 69:698–709. https://doi.org/10.1007/s00248-014-0524-0Archibald PA (1977) Physiological characteristics of Trebouxia (Chlorophyceae, Chlorococcales) and Pseudotrebouxia (Chlorophyceae, Chlorosarcinales). Phycologia 16:295–300. https://doi.org/10.2216/i0031-8884-16-3-295.1Armstrong RA (2017) Adaptation of lichens to extreme conditions. In: Kumar V, Shukla S, Kumar N (eds) Plant adaptation strategies in changing environment. Springer Singapore, Singapore, pp 1–27Arup U (1995) Littoral species of Caloplaca in North America: a summary and a key. Bryologist 98:129–140. https://doi.org/10.2307/3243649Aschenbrenner IA, Cernava T, Berg G, Grube M (2016) Understanding microbial multi-species symbioses. Front Microbiol 7:180. https://doi.org/10.3389/fmicb.2016.00180Balarinová K, Barták M, Hazdrová J, Hájek J, Jílková J (2014) Changes in photosynthesis, pigment composition and glutathione contents in two Antarctic lichens during a light stress and recovery. Photosynthetica 52:538–547. https://doi.org/10.1007/s11099-014-0060-7Biosca EG, Flores R, Santander RD, Díez-Gil JL, Barreno E (2016) Innovative approaches using lichen enriched media to improve isolation and culturability of lichen associated bacteria. PLoS One 11:e0160328. https://doi.org/10.1371/journal.pone.0160328Bischoff HW, Bold HC (1963) Some soil algae from Enchanted Rock and related algal species. Phycol Stud 44(1):1–95Borges L, Caldas S, Montes D’Oca MG, Abreu PC (2016) Effect of harvesting processes on the lipid yield and fatty acid profile of the marine microalga Nannochloropsis oculata. Aquac Rep 4:164–168. https://doi.org/10.1016/j.aqrep.2016.10.004Brandt A, Posthoff E, de Vera J-P, Onofri S, Ott S (2016) Characterisation of growth and ultrastructural effects of the Xanthoria elegans photobiont after 1.5 years of space exposure on the International Space Station. Orig Life Evol Biosph 46:311–321. https://doi.org/10.1007/s11084-015-9470-1Brányiková I, Maršálková B, Doucha J et al (2011) Microalgae—novel highly efficient starch producers. Biotechnol Bioeng 108:766–776. https://doi.org/10.1002/bit.23016Callis J, Carpenter T, Sun CW, Vierstra RD (1995) Structure and evolution of genes encoding polyubiquitin and ubiquitin-like proteins in Arabidopsis thaliana ecotype Columbia. Genetics 139:921–939Campenni L, Nobre BP, Santos CA et al (2013) Carotenoid and lipid production by the autotrophic microalga Chlorella protothecoides under nutritional, salinity, and luminosity stress conditions. Appl Microbiol Biotechnol 97:1383–1393. https://doi.org/10.1007/s00253-012-4570-6Casano LM, del Campo EM, García-Breijo FJ et al (2011) Two Trebouxia algae with different physiological performances are ever-present in lichen thalli of Ramalina farinacea. Coexistence versus competition? Environ Microbiol 13:806–818. https://doi.org/10.1111/j.1462-2920.2010.02386.xChettri M, Cook C, Vardaka E, Sawidis T, Lanaras L (1998) The effect of Cu, Zn and Pb on the chlorophyll content of the lichens Cladonia convoluta and Cladonia rangiformis. Environ Exp Bot 39:1–10. https://doi.org/10.1016/S0098-8472(97)00024-5Cornillon P-A (2012) R for statistics. CRC Press, Boca RatonCowan AK, Rose PD, Horne LG (1992) Dunaliella salina: a model system for studying the response of plant cells to stress. J Exp Bot 43:1535–1547. https://doi.org/10.1093/jxb/43.12.1535Czechowski T, Stitt M, Altmann T, Udvardi MK, Scheible WR (2005) Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol 139:5–17. https://doi.org/10.1104/pp.105.063743Danquah A, de Zelicourt A, Colcombet J, Hirt H (2014) The role of ABA and MAPK signaling pathways in plant abiotic stress responses. Biotechnol Adv 32:40–52. https://doi.org/10.1016/j.biotechadv.2013.09.006Delmail D, Labrousse P, Hourdin P et al (2013) Micropropagation of Myriophyllum alterniflorum (Haloragaceae) for stream rehabilitation: first in vitro culture and reintroduction assays of a heavy-metal hyperaccumulator immersed macrophyte. Int J Phytoremediation 15:647–662. https://doi.org/10.1080/15226514.2012.723068Dragone G, Fernandes BD, Abreu AP, Vicente AA, Teixeira JA (2011) Nutrient limitation as a strategy for increasing starch accumulation in microalgae. Appl Energy 88:3331–3335. https://doi.org/10.1016/j.apenergy.2011.03.012Duarte AWF, Passarini MRZ, Delforno TP et al (2016) Yeasts from macroalgae and lichens that inhabit the South Shetland Islands, Antarctica. Environ Microbiol Rep 8:874–885. https://doi.org/10.1111/1758-2229.12452Durgbanshi A, Arbona V, Pozo O et al (2005) Simultaneous determination of multiple phytohormones in plant extracts by liquid chromatography–electrospray tandem mass spectrometry. J Agric Food Chem 53:8437–8442. https://doi.org/10.1021/JF050884BEinspahr KJ, Maeda M, Thompson GA (1988) Concurrent changes in Dunaliella salina ultrastructure and membrane phospholipid metabolism after hyperosmotic shock. J Cell Biol 107:529–538. https://doi.org/10.1083/JCB.107.2.529Gasulla F, de Nova PG, Esteban-Carrasco A et al (2009) Dehydration rate and time of desiccation affect recovery of the lichenic algae Trebouxia erici: alternative and classical protective mechanisms. Planta 231:195–208. https://doi.org/10.1007/s00425-009-1019-yGasulla F, Guéra A, Barreno E (2010) A simple and rapid method for isolating lichen photobionts. Symbiosis 51:175–179. https://doi.org/10.1007/s13199-010-0064-4Gómez-Cadenas A, Arbona V, Jacas J, Primo-Millo E, Talon M (2002) Abscisic acid reduces leaf abscission and increases salt tolerance in citrus plants. J Plant Growth Regul 21:234–240. https://doi.org/10.1007/s00344-002-0013-4Green TGA, Brabyn L, Beard C, Sancho LG (2012) Extremely low lichen growth rates in Taylor Valley, Dry Valleys, continental Antarctica. Polar Biol 35:535–541. https://doi.org/10.1007/s00300-011-1098-7Grube M, Blaha J (2005) Halotolerance and lichen symbioses. In: Gunde-Cimerman N, Oren A, Plemenitaš A (eds) Adaptation to life at high salt concentrations in Archaea, Bacteria, and Eukarya. Springer, Berlin, pp 471–488Guéra A, Calatayud A, Sabater B, Barreno E (2004) Involvement of the thylakoidal NADH-plastoquinone-oxidoreductase complex in the early responses to ozone exposure of barley (Hordeum vulgare L.) seedlings. J Exp Bot 56:205–218. https://doi.org/10.1093/jxb/eri024Gustavs L, Eggert A, Michalik D, Karsten U (2010) Physiological and biochemical responses of green microalgae from different habitats to osmotic and matric stress. Protoplasma 243:3–14. https://doi.org/10.1007/s00709-009-0060-9Hauser F, Rainer W, Schroeder JI (2011) Evolution of abscisic acid synthesis and signaling mechanisms. Curr Biol 21:346–355. https://doi.org/10.1016/j.cub.2011.03.015.HauserHayashi H, Alia L, Mustardy L, Ida M, Murata N (1997) Transformation of Arabidopsis thaliana with the codA gene for choline oxidase; accumulation of glycine betaine and enhanced tolerance to salt and cold stress. Plant J 12:133–142. https://doi.org/10.1046/j.1365-313X.1997.12010133.xHiremath S, Mathad P (2010) Impact of salinity on the physiological and biochemical traits of Chlorella vulgaris Beijerinck. J Algal Biomass Util 1:51–59Hirsch R, Hartung W, Gimmler H (1989) Abscisic acid content of algae under stress. Bot Acta 102:326–334. https://doi.org/10.1111/j.1438-8677.1989.tb00113.xJameson P (1993) Plant hormones in the algae. Prog Phycol Res 9:239–279Kibbe WA (2007) OligoCalc: an online oligonucleotide properties calculator. Nucleic Acids Res 35:W43–W46. https://doi.org/10.1093/nar/gkm234Kirk PM, Cannon PF, David JC, Stalpers JA (2001) Ainsworth and Bisby’s dictionary of the fungi, 9th edn. CABI Publishing, Wallingford, UKKline KG, Barrett-Wilt GA, Sussman MR (2010) In planta changes in protein phosphorylation induced by the plant hormone abscisic acid. Proc Natl Acad Sci USA 107:15986–15991. https://doi.org/10.1073/pnas.1007879107Koizumi M, Yamaguchi-Shinozaki K, Tsuji H, Shinozaki K (1993) Structure and expression of two genes that encode distinct drought-inducible cysteine proteinases in Arabidopsis thaliana. Gene 129:175–182. https://doi.org/10.1016/0378-1119(93)90266-6Krause GH, Weis E (1991) Chlorophyll fluorescence and photosynthesis: the basics. Annu Rev Plant Physiol Plant Mol Biol 42:313–349. https://doi.org/10.1146/annurev.pp.42.060191.001525Lan SB, Wu L, Zhang DL, Hu CX, Liu YD (2010) Effects of drought and salt stresses on man-made cyanobacterial crusts. Eur J Soil Biol 46:381–386. https://doi.org/10.1016/j.ejsobi.2010.08.002Leavitt SD, Kraichak E, Nelsen MP et al (2015) Fungal specificity and selectivity for algae play a major role in determining lichen partnerships across diverse ecogeographic regions in the lichen-forming family Parmeliaceae (Ascomycota). Mol Ecol 24:3779–3797. https://doi.org/10.1111/mec.13271Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/METH.2001.1262Lu Y, Xu J (2015) Phytohormones in microalgae: a new opportunity for microalgal biotechnology? Trends Plant Sci 20:273–282. https://doi.org/10.1016/j.tplants.2015.01.006Malaspina P, Giordani P, Pastorino G, Modenesi P, Mariotti MG (2015) Interaction of sea salt and atmospheric pollution alters the OJIP fluorescence transient in the lichen Pseudevernia furfuracea (L.) Zopf. Ecol Indic 50:251–257. https://doi.org/10.1016/j.ecolind.2014.11.015Mane AV, Karadge BA, Samant JS (2010) Salt stress induced alteration in photosynthetic pigments and polyphenols of Pennisetum alopecuroides (L.). J Ecophysiol Occup Health 10:177–182. https://doi.org/10.18311/jeoh/2010/18339Maphangwa KW, Musil CF, Raitt L, Zedda L (2012) Experimental climate warming decreases photosynthetic efficiency of lichens in an arid South African ecosystem. Oecologia 169:257–268. https://doi.org/10.1007/s00442-011-2184-9Margulis L, Barreno E (2003) Looking at lichens. Bioscience 53:776–778. https://doi.org/10.1641/0006-3568(2003)053%5b0776:lal%5d2.0.co;2Maršálek B, Zahradníčková H, Hronková M (1992) Extracellular abscisic acid produced by cyanobacteria under salt stress. J Plant Physiol 139:506–508. https://doi.org/10.1016/S0176-1617(11)80503-1Martínez-Alberola F (2015) Genome characterization of the symbiotic microalga Trebouxia sp. TR9 isolated from the lichen Ramalina farinacea (L.) Ach. by means of NGS techniques. PhD Dissertation. Universitat de València. http://roderic.uv.es/handle/10550/48824Mishra A, Jha B (2009) Isolation and characterization of extracellular polymeric substances from micro-algae Dunaliella salina under salt stress. Bioresour Technol 100:3382–3386. https://doi.org/10.1016/j.biortech.2009.02.006Molins A, Moya P, García-Breijo FJ, Reig-Arminana J, Barreno E (2018) A multi-tool approach to assess microalgal diversity in lichens: isolation, Sanger sequencing, HTS and ultrastructural correlations. Lichenologist 50:123–138. https://doi.org/10.1017/S0024282917000664Moya P, Molins A, Martínez-Alberola F, Muggia L, Barreno E (2017) Unexpected associated microalgal diversity in the lichen Ramalina farinacea is uncovered by pyrosequencing analyses. PLoS One 12:e0175091. https://doi.org/10.1371/journal.pone.0175091Nash TH III, Lange OL (1988) Responses of lichens to salinity: concentration and time-course relationships and variability among Californian species. New Phytol 109:361–367. https://doi.org/10.1111/j.1469-8137.1988.tb04206.xNeale PJ, Melis A (1989) Salinity-stress enhances photoinhibition of photosynthesis in Chlamydomonas reinhardtii. J Plant Physiol 134:619–622. https://doi.org/10.1016/S0176-1617(89)80158-0Negrão S, Schmöckel SM, Tester M (2017) Evaluating physiological responses of plants to salinity stress. Ann Bot 119:1–11. https://doi.org/10.1093/aob/mcw191Qiao K, Takano T, Liu S (2015) Discovery of two novel highly tolerant NaHCO3 Trebouxiophytes: identification and characterization of microalgae from extreme saline–alkali soil. Algal Res 9:245–253. https://doi.org/10.1016/j.algal.2015.03.023Ruzin SE (2000) Plant microtechnique and microscopy. New Phytol 148:57–58Schwartz SH, Tan BC, Gage DA, Zeevaart JAD, McCarty DR (1997) Specific oxidative cleavage of carotenoids by VP14 of maize. Science 276:1872–1874. https://doi.org/10.1126/science.276.5320.1872Škaloud P, Peksa O (2010) Evolutionary inferences based on ITS rDNA and actin sequences reveal extensive diversity of the common lichen alga Asterochloris (Trebouxiophyceae, Chlorophyta). Mol Phylogenet Evol 54:36–46. https://doi.org/10.1016/J.YMPEV.2009.09.035Spribille T, Tuovinen V, Resl P et al (2016) Basidiomycete yeasts in the cortex of ascomycete macrolichens. Science 353:488–492. https://doi.org/10.1126/science.aaf8287Stepien P, Johnson GN (2009) Contrasting responses of photosynthesis to salt stress in the glycophyte Arabidopsis and the halophyte Thellungiella: role of the plastid terminal oxidase as an alternative electron sink. Plant Physiol 149:1154–1165. https://doi.org/10.1104/pp.108.132407Takagi M, Karseno YT (2006) Effect of salt concentration on intracellular accumulation of lipids and triacylglyceride in marine microalgae Dunaliella cells. J Biosci Bioeng 101:223–226. https://doi.org/10.1263/jbb.101.223Takahagi T, Yamamoto Y, Kinoshita Y, Takeshita S, Yamada T (2002) Inhibitory effects of sodium chloride on induction of tissue cultures of lichens of Ramalina species. Plant Biotechnol 19:53–55. https://doi.org/10.5511/plantbiotechnology.19.53Tietz A, Kasprik W (1986) Identification of abscisic acid in a green alga. Biochem Physiol Pflanz 181:269–274. https://doi.org/10.1016/S0015-3796(86)80093-2Wani SH, Kumar V, Shriram V, Sah SK (2016) Phytohormones and their metabolic engineering for abiotic stress tolerance in crop plants. Crop J 4:162–176. https://doi.org/10.1016/j.cj.2016.01.010Wellburn AR, Lichtenthaler H (1984) Formulae and program to determine total carotenoids and chlorophylls A and B of leaf extracts in different solvents. In: Sybesma C (ed) Advances in photosynthesis research. Springer, Dordrecht, pp 9–12Zhang J, Jia W, Yang J, Ismail AM (2006) Role of ABA in integrating plant responses to drought and salt stresses. Field Crop Res 97:111–119. https://doi.org/10.1016/j.fcr.2005.08.01

Fitting Carbon Gels and Composites for Environmental Processes

International audienc