Genetic variation in populations of the earthworm, Lumbricus rubellus, across contaminated mine sites

Background  Populations of the earthworm, Lumbricus rubellus, are commonly found across highly contaminated former mine sites and are considered to have under-gone selection for mitigating metal toxicity. Comparison of adapted populations with those found on less contaminated soils can provide insights into ecological processes that demonstrate the long-term effects of soil contamination. Contemporary sequencing methods allow for portrayal of demographic inferences and highlight genetic variation indicative of selection at specific genes. Furthermore, the occurrence of L. rubellus lineages across the UK allows for inferences of mechanisms associated with drivers of speciation and local adaptation.  Results  Using RADseq, we were able to define population structure between the two lineages through the use of draft genomes for each, demonstrating an absence of admixture between lineages and that populations over extensive geographic distances form discrete populations. Between the two British lineages, we were able to provide evidence for selection near to genes associated with epigenetic and morphological functions, as well as near a gene encoding a pheromone. Earthworms inhabiting highly contaminated soils bare close genomic resemblance to those from proximal control soils. We were able to define a number of SNPs that largely segregate populations and are indicative of genes that are likely under selection for managing metal toxicity. This includes calcium and phosphate-handling mechanisms linked to lead and arsenic contaminants, respectively, while we also observed evidence for glutathione-related mechanisms, including metallothionein, across multiple populations. Population genomic end points demonstrate no consistent reduction in nucleotide diversity, or increase in inbreeding coefficient, relative to history of exposure.   Conclusions  Though we can clearly define lineage membership using genomic markers, as well as population structure between geographic localities, it is difficult to resolve markers that segregate entirely between populations in response to soil metal concentrations. This may represent a highly variable series of traits in response to the heterogenous nature of the soil environment, but ultimately demonstrates the maintenance of lineage-specific genetic variation among local populations. L. rubellus appears to provide an exemplary system for exploring drivers for speciation, with a continuum of lineages coexisting across continental Europe, while distinct lineages exist in isolation throughout the UK

Characterizing the involvement of FaMADS9 in the regulation of strawberry fruit receptacle development

FaMADS9 is the strawberry (Fragaria x ananassa) gene that exhibits the highest homology to the tomato (Solanum lycopersicum) RIN gene. Transgenic lines were obtained in which FaMADS9 was silenced. The fruits of these lines did not show differences in basic parameters, such as fruit firmness or colour, but exhibited lower Brix values in three of the four independent lines. The gene ontology MapMan category that was most enriched among the differentially expressed genes in the receptacles at the white stage corresponded to the regulation of transcription, including a high percentage of transcription factors and regulatory proteins associated with auxin action. In contrast, the most enriched categories at the red stage were transport, lipid metabolism and cell wall. Metabolomic analysis of the receptacles of the transformed fruits identified significant changes in the content of maltose, galactonic acid-1,4-lactone, proantho- cyanidins and flavonols at the green/white stage, while isomaltose, anthocyanins and cuticular wax metabolism were the most affected at the red stage. Among the regulatory genes that were differentially expressed in the transgenic receptacles were several genes previously linked to flavonoid metabolism, such as MYB10, DIV, ZFN1, ZFN2, GT2, and GT5, or associated with the action of hormones, such as abscisic acid, SHP, ASR, GTE7 and SnRK2.7. The inference of a gene regulatory network, based on a dynamic Bayesian approach, among the genes differentially expressed in the transgenic receptacles at the white and red stages, identified the genes KAN1, DIV, ZFN2 and GTE7 as putative targets of FaMADS9. A MADS9-specific CArG box was identified in the promoters of these genes

Study of the Falling Friction Effect on Rolling Contact Parameters

[EN] The existence of a wheel rail friction coefficient that depends on the slip velocity has been associated in the literature with important railway problems like the curving squeal and certain corrugation problems in rails. Rolling contact models that take into account this effect were carried out through the so-called Exact Theories adopting an exact elastic model of the solids in contact, and Simplified Theories which assume simplified elastic models such as Winkler. The former ones, based on Kalker s Variational Theory, give rise to numerical problems; the latter ones need to adopt hypotheses that significantly deviate from actual conditions, leading to unrealistic solutions of the contact problem. In this paper, a methodology based on Kalker s Variational Theory is presented, in which a local slip velocity-dependent friction law is considered. A formulation to get steady-state conditions of rolling contact by means of regularisation of the Coulomb s law is proposed. The model allows establishing relationships in order to estimate the global properties (creepage velocities vs. total longitudinal forces) through local properties (local slip velocity vs. coefficient of friction) or vice versa. The proposed model shows a good agreement with experimental tests while solving the numerical problems previously mentioned.The authors acknowledge the financial contribution of the Spanish Ministry of Economy and Competitiveness through the Project TRA2013-45596-C2-1-R.Giner Navarro, J.; Baeza González, LM.; Vila Tortosa, MP.; Alonso Pazos, A. (2017). Study of the Falling Friction Effect on Rolling Contact Parameters. Tribology Letters. 65(1). https://doi.org/10.1007/s11249-016-0810-8S651Grassie, S.L., Elkins, J.A.: Rail corrugation on North American transit systems. Veh. Syst. Dyn. 28, 5–17 (1998)Hsu, S.S., Huang, Z., Iwnicki, S.D., Thompson, D.J., Jones, C.J.C., Xie, G., Allen, P.D.: Experimental and theoretical investigation of railway wheel squeal. Proc. Inst. Mech. Eng. F J. Rail Rapid Transit 221, 59–73 (2007)Kalker, J.J.: Three-Dimensional Elastic Bodies in Rolling Contact. Kluwer, Dordrecht (1990)Polach, O.: Influence of locomotive tractive effort on the forces between wheel and rail. Veh. Syst. Dyn. 35, 7–22 (2001)Giménez, J.G., Alonso, A., Gómez, E.: Introduction of a friction coefficient dependent on the slip in the FastSim algorithm. Veh. Syst. Dyn. 43, 233–244 (2005)Baeza, L., Vila, P., Roda, A., Fayos, J.: Prediction of corrugation in rails using a non-stationary wheel–rail contact model. Wear 265, 1156–1162 (2008)Vollebregt, E.A.H., Schuttelaars, H.M.: Quasi-static analysis of two-dimensional rolling contact with slip-velocity dependent friction. J. Sound Vib. 331, 2141–2155 (2012)Avlonitis, M., Kalaitzidou, K., Streator, J.: Investigation of friction statics and real contact area by means a modified OFC model. Tribol. Int. 69, 168–175 (2014)Berger, E.J., Mackin, T.J.: On the walking stick–slip problem. Tribol. Int. 75, 51–60 (2014)Alonso, A., Guiral, A., Baeza, B., Iwnicki, S.D.: Wheel–rail contact: experimental study of the creep forces–creepage relationships. Veh. Syst. Dyn. 52(S1), 469–487 (2014)Spiryagin, M., Polach, O., Cole, C.: Creep force modelling for rail traction vehicles based on the Fastsim algorithm. Veh. Syst. Dyn. 51, 1765–1783 (2013)Vollebregt, E.A.H.: Numerical modeling of measured railway creep versus creep-force curves with CONTACT. Wear 314, 87–95 (2014)Kalker, J.J.: On the Rolling Contact of Two Elastic Bodies in the Presence of Dry Friction. PhD Thesis, Technical University of Delft (Holland) (1967)Baeza, L., Fuenmayor, F.J., Carballeira, J., Roda, A.: Influence of the wheel–rail contact instationary process on contact parameters. J. Strain Anal. Eng. 42, 377–387 (2007)Le Rouzic, J., Le Bot, A., Perret-Liaudet, J., Guibert, M., Rusanov, A., Douminge, L., Bretagnol, F., Mazuyer, D.: Friction-induced vibration by Stribeck’s law: application to wiper blade squeal noise. Tribol. Lett. 49, 563–572 (2013)Rabinowicz, E.: The nature of the static and kinetic coefficients of friction. J. Appl. Phys. 22, 1373–1379 (1951)Carter, F.W.: On the action of locomotive driving wheel. Proc. R. Soc. Lon. Ser. A 112, 151–157 (1926)Kalker, J.J.: A fast algorithm for the simplified theory of rolling contact. Veh. Syst. Dyn. 11, 1–13 (1982

Response of the photosynthetic apparatus to a flowering-inductive period by water stress in Citrus

The photosynthetic responses to a flowering-inductive water-stress period and recovery were studied and compared in two Citrus species. Under greenhouse conditions, Fino lemon and Owari satsuma trees were subjected to moderate (-2 MPa at predawn) and severe (-3 MPa) water stress levels and were re-watered after 60 days. Vegetative growth was inhibited during the stress assays, and strong defoliation levels were reported, especially in Fino lemon. In both species, bud sprouting was induced after re-watering. Flowers and vegetative shoots developed in Owari satsuma after a drought period, and the development was independent of the stress level. In Fino lemon, vegetative shoots and flowers were primarily formed after moderate and severe stress, respectively. The photosynthetic rate and stomatal conductance were reduced by water stress, and a marked increase in water-use efficiency at the moderate water deficit level was observed. Nevertheless, the photosynthetic apparatus was not damaged, since the maximum quantum yield, photosynthetic pigment concentrations and Rubisco level and activity did not change. Furthermore, the measured malonyldialdehyde (MDA) and peroxidase activity indicated that oxidative stress was not specifically triggered by water stress in our study. Therefore, the gas exchange, fluorescence and biochemical parameters suggested that diffusional limitations to photosynthesis predominated in both of the studied Citrus species, and explained the rapid recovery of the photosynthetic parameters after rehydration. The net CO 2 fixation rate and stomatal conductance were recovered within 24 h in Fino lemon, whereas 3 days were required in Owari satsuma. This suggests the presence of some metabolic limitations in the latter species. Furthermore, the sensibility of the defoliation rates, the accumulation of proline and the stomatal behaviour in response to water stress indicated a higher drought tolerance of Fino lemon, according to its better acclimation to hot climates. © 2011 Springer-Verlag.The authors thank Dr. J. Moreno and co-workers from the Departamento de Bioquimica of the Universidad de Valencia for his help and support in the Rubisco assays, and Dr. F. Fornes, Dr. A. Calatayud and Dr. E. Primo-Millo for the critical review of the manuscript. This work was funded by the Universitat Politecnica de Valencia, Spain (Ayudas para primeros proyectos de investigacion PAID06-06).Ávila Reséndiz, C.; Guardiola Barcena, JL.; González Nebauer, S. (2012). Response of the photosynthetic apparatus to a flowering-inductive period by water stress in Citrus. Trees - Structure and Function. 26(3):833-840. https://doi.org/10.1007/s00468-011-0657-4S833840263Addicott FT (1982) Abscission. University of California Press, BerkeleyBajji M, Kinet JM, Lutts S (1998) Salt stress effects on roots and leaves of Atriplex halimus L. and their corresponding callus. Plant Sci 137:131–142Barbera G, Fatta-del-Bosco G, Lo-Cascio B (1985) Effect of water stress on lemon summer bloom: the Forzatura technique in the Sicilian citrus industry. Acta Hortic 171:391–397Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207Bota J, Medrano H, Flexas J (2004) Is photosynthesis limited by decreased Rubisco activity and RuBP content under progressive water stress? New Phytol 162:671–681Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254Cassin J, Bourdeaut A, Fougue V, Furon V, Gaillard JP, LeBourdelles J, Montagut G, Moreuil C (1969) The influence of climate upon blooming of Citrus in tropical areas. Proc Int Soc Citrus 1:315–323Castel JR, Buj A (1990) Response of Salustiana oranges to high frequency deficit irrigation. Irrig Sci 11:121–127Chaikiatitiyos S, Menzel CM, Rasmussen TS (1994) Floral induction in tropical fruit trees: effects of temperature and water supply. J Hortic Sci 69:397–415Chaves MM, Flexas J, Pinheiro C (2009) Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. Ann Bot 103:551–560Costa JM, Ortuño MF, Chaves M (2007) Deficit irrigation as a strategy to save water: physiology and potential application to horticulture. J Integr Plant Biol 49:1421–1434Davenport TL (1990) Citrus flowering. Hortic Rev 12:249–408Davies FS, Albrigo LG (1994) Citrus. CAB International, Wallingford, pp 126–134Domingo R, Ruiz-Sánchez MC, Sánchez-Blanco MJ, Torrecillas A (1996) Water relations, growth and yield of Fino lemon trees under regulated deficit irrigation. Irrig Sci 16:115–123Erismann ND, Machado EC, Tucci MLS (2008) Photosynthetic limitation by CO2 diffusion in drought stressed orange leaves on three rootstocks. Photosynth Res 96:163–172Flexas J, Bota J, Galmés J, Medrano H, Ribas-Carbó M (2006) Keeping a positive carbon balance under adverse conditions: responses of photosynthesis and respiration to water stress. Physiol Plant 127:343–352Gallé A, Florez-Sarasa I, Tomas M, Pou A, Medrano H, Ribas-Carbó M, Flexas J (2009) The role of mesophyll conductance during water stress and recovery in tobacco (Nicotiana sylvestris): acclimation or limitation? J Exp Bot 60:2379–2390Galmés J, Medrano H, Flexas J (2007) Photosynthetic limitations in response to water stress and recovery in Mediterrenean plants with different growth forms. New Phytol 175:81–93García-Luis A, Kanduser M, Santamarina P, Guardiola JL (1992) Low temperature influence on flowering in Citrus. The separation of inductive and bud dormancy releasing effects. Physiol Plant 86:648–652García-Sánchez F, Syvertsen JP, Gimeno V, Botía P, Pérez-Pérez JG (2007) Responses to flooding and drought stress by two citrus rootstock seedlings with different water-use efficiency. Physiol Plant 130:532–542Genty B, Briantais JM, Baker NR (1989) The relationship between quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim Biophys Acta 990:87–92Gómez-Cadenas A, Tadeo FR, Talon M, Primo-Millo E (1996) Leaf abscission induced by ethylene in water-stressed intact seedlings of Cleopatra mandarin requires previous abscisic acid accumulation in roots. Plant Physiol 112:401–408Gordo O, Sanz JJ (2010) Impact of climate change on plant phenology in Mediterranean ecosystems. Glob Chang Biol 16:1082–1106Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–190Hoagland DR, Arnon DI (1950) The water culture method for growing plants without soil. California Agricultural Experiment Station Circular no. 347, p 32IPCC (2001) Climate change 2001. In: Houghton JT (ed) The scientific basis. Cambridge University Press, CambridgeLawlor DW (1995) The effects of water deficit on photosynthesis. In: Smirnoff N (ed) Environment and plant metabolism. Bios Scientific Publishers, Oxford, pp 129–160Lichtenthaler HK, Buschmann C (2001) Current protocols in food analytical chemistry, F4.2.1 and F4.3.1. John Wiley and Sons, Inc, NJLorimer GH, Badger MR, Andrews TJ (1977) D-Ribulose-1, 5-bisphosphate carboxilase-oxigenase. Improved methods for activation and assay of catalytic activities. Anal Biochem 78:66–75Miyashita K, Tanakamaru S, Maitani T, Kimura K (2005) Recovery responses of photosynthesis, transpiration, and stomatal conductance in kidney bean following drought stress. Environ Exp Bot 53:205–214Nir I, Leshem B, Goren R (1972) Effects of water stress, gibberellic acid and 2-chloroethylammoniumchloride (CCC) ob flower differentiation in Eureka lemon trees. J Am Soc Hortic Sci 97:774–778Peñarrubia L, Moreno J (1988) Ribulose 1, 5-bisphosphate carboxylase oxygenase from citrus leaves. Phytochemistry 27:1999–2004Pérez-Pérez JG, Syvertsen JP, Botía P, García-Sánchez F (2007) Leaf water relations and net gas exchange responses of salinized carrizo citrange seedlings during drought stress and recovery. Ann Bot 100:335–345Pérez-Pérez JG, Robles JM, Tovar JC, Botía P (2009) Response to drought and salt stress of lemon ‘Fino 49’ under field conditions: water relations, osmotic adjustment and gas Exchange. Sci Hortic 122:83–90Reynolds M, Tuberosa R (2008) Translational research impacting on crop productivity in drought-prone environments. 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Frequent Arousal from Hibernation Linked to Severity of Infection and Mortality in Bats with White-Nose Syndrome

White-nose syndrome (WNS), an emerging infectious disease that has killed over 5.5 million hibernating bats, is named for the causative agent, a white fungus (Geomyces destructans (Gd)) that invades the skin of torpid bats. During hibernation, arousals to warm (euthermic) body temperatures are normal but deplete fat stores. Temperature-sensitive dataloggers were attached to the backs of 504 free-ranging little brown bats (Myotis lucifugus) in hibernacula located throughout the northeastern USA. Dataloggers were retrieved at the end of the hibernation season and complete profiles of skin temperature data were available from 83 bats, which were categorized as: (1) unaffected, (2) WNS-affected but alive at time of datalogger removal, or (3) WNS-affected but found dead at time of datalogger removal. Histological confirmation of WNS severity (as indexed by degree of fungal infection) as well as confirmation of presence/absence of DNA from Gd by PCR was determined for 26 animals. We demonstrated that WNS-affected bats aroused to euthermic body temperatures more frequently than unaffected bats, likely contributing to subsequent mortality. Within the subset of WNS-affected bats that were found dead at the time of datalogger removal, the number of arousal bouts since datalogger attachment significantly predicted date of death. Additionally, the severity of cutaneous Gd infection correlated with the number of arousal episodes from torpor during hibernation. Thus, increased frequency of arousal from torpor likely contributes to WNS-associated mortality, but the question of how Gd infection induces increased arousals remains unanswered

Neurons of the Dentate Molecular Layer in the Rabbit Hippocampus

The molecular layer of the dentate gyrus appears as the main entrance gate for information into the hippocampus, i.e., where the perforant path axons from the entorhinal cortex synapse onto the spines and dendrites of granule cells. A few dispersed neuronal somata appear intermingled in between and probably control the flow of information in this area. In rabbits, the number of neurons in the molecular layer increases in the first week of postnatal life and then stabilizes to appear permanent and heterogeneous over the individuals’ life span, including old animals. By means of Golgi impregnations, NADPH histochemistry, immunocytochemical stainings and intracellular labelings (lucifer yellow and biocytin injections), eight neuronal morphological types have been detected in the molecular layer of developing adult and old rabbits. Six of them appear as interneurons displaying smooth dendrites and GABA immunoreactivity: those here called as globoid, vertical, small horizontal, large horizontal, inverted pyramidal and polymorphic. Additionally there are two GABA negative types: the sarmentous and ectopic granular neurons. The distribution of the somata and dendritic trees of these neurons shows preferences for a definite sublayer of the molecular layer: small horizontal, sarmentous and inverted pyramidal neurons are preferably found in the outer third of the molecular layer; vertical, globoid and polymorph neurons locate the intermediate third, while large horizontal and ectopic granular neurons occupy the inner third or the juxtagranular molecular layer. Our results reveal substantial differences in the morphology and electrophysiological behaviour between each neuronal archetype in the dentate molecular layer, allowing us to propose a new classification for this neural population

New structural insights into the role of TROVE2 complexes in the on-set and pathogenesis of systemic lupus eythematosus determined by a combiantion of QCM-D and DPI

The final publication is available at link.springer.com.[EN] The mechanism of self-recognition of the autoantigen TROVE2, a common biomarker in autoimmune diseases, has been studied with a quartz crystal microbalance with dissipation monitoring (QCM-D) and dual polarization interferometry (DPI). The complementarity and remarkable analytical features of both techniques has allowed new insights into the onset of systemic lupus erythematosus (SLE) to be achieved at the molecular level. The in vitro study for SLE patients and healthy subjects suggests that anti-TROVE2 autoantibodies may undergo an antibody bipolar bridging. An epitope-paratope-specific binding initially occurs to activate a hidden Fc receptor in the TROVE2 tertiary structure. This bipolar mechanism may contribute to the pathogenic accumulation of anti-TROVE2 autoantibody immune complex in autoimmune disease. Furthermore, the specific calcium-dependent protein-protein bridges point out at how the TRIM21/TROVE2 association might occur, suggesting that the TROVE2 protein could stimulate the intracellular immune signaling via the TRIM21 PRY-SPRY domain. These findings may help to better understand the origins of the specificity and affinity of TROVE2 interactions, which might play a key role in the SLE pathogenesis. This manuscript gives one of the first practical applications of two novel functions (-df/dD and Delta h/molec) for the analysis of the data provided by QCM-D and DPI. In addition, it is the first time that QCM-D has been used for mapping hidden Fc receptors as well as linear epitopes in a protein tertiary structure.We would like to thank Sylvia Daunert for her invaluable help with the discussion of the paper. Furthermore, we acknowledge financial support from the Generalitat Valenciana (GVA-PROMETEOII/2014/040) as well as the Spanish Ministry of Economy and Competitiveness and the European Regional Development Fund under award numbers CTQ2013-45875-R and CTQ2013-42914-RJuste-Dolz, AM.; Do Nascimento, NM.; Monzó, IS.; Grau-García, E.; Roman-Ivorra, JA.; López-Paz, JL.; Escorihuela Fuentes, J.... (2019). New structural insights into the role of TROVE2 complexes in the on-set and pathogenesis of systemic lupus eythematosus determined by a combiantion of QCM-D and DPI. Analytical and Bioanalytical Chemistry. 411(19):4709-4720. https://doi.org/10.1007/s00216-018-1407-xS4709472041119Kakatia S, Teronpia R, Barmanb B. Frequency, pattern and determinants of flare in systemic lupus erythematosus: a study from North East India. Egypt Rheumatol. 2015;37:S55–9.Kuhn A, Wenzel J, Weyd H. Photosensitivity, apoptosis, and cytokines in the pathogenesis of lupus erythematosus: a critical review. 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Stress-induced c-Fos expression is differentially modulated by dexamethasone, diazepam and imipramine

Immobilization stress upregulates c-Fos expression in several CNS areas. Repeated stress or the use of drugs can modulate stress-induced c-Fos expression. Here, we investigated in 40 different areas of the rat brain the effects of dexamethasone (SDX, a synthetic glucocorticoid), diazepam (SBDZ, a benzodiazepine), and imipramine (IMI, an antidepressant) on the c-Fos expression induced by restraint stress. Wistar rats were divided into four groups and submitted to 20 days of daily injection of saline (three first groups) or imipramine, 15 mg/kg, i.p. On day 21, animals were submitted to injections of saline (somatosensory, SS), SDX (1 mg/kg, i.p.), SBDZ (5 mg/kg, i.p.), or IMI (15 mg/kg, i.p.) before being submitted to restraint. Immediately after stress, the animals were perfused and their brains processed with immunohistochemistry for c-Fos (Ab-5 Oncogene Science). Dexamethasone reduced stress- induced c-Fos expression in SS cortex, hippocampus, paraventricular nucleus of the hypothalamus (PVH), and locus coeruleus (LC), whereas diazepam reduced c-Fos staining in the SS cortex, hippocampus, bed nucleus of stria terminalis, septal area, and hypothalamus (preoptic area and supramammillary nucleus). Chronic administration of imipramine decreased staining in the hippocampus, PVH, and LC, while increasing it in the nucleus raphe pallidus. We conclude that dexamethasone, diazepam and imipramine differentially modulate stress-induced Fos expression. the present study provides an important comparative background that may help in the further understanding of the effects of these compounds and on the brain activation as well as on the behavioral, neuroendocrine, and autonomic responses to stress.UFRRJ, Dept Physiol Sci, BR-23890000 Rio de Janeiro, BrazilUniversidade Federal de São Paulo, Dept Physiol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Psychobiol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Physiol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Psychobiol, São Paulo, BrazilWeb of Scienc