On the relationship between C and N fixation in nodulated alfalfa (Medicago sativa)

Legumes such as alfalfa (Medicago sativa L.) are vital N2-fixing crops accounting for a global N2 fixation of ~35 MtNyear-1. Although enzymatic and molecular mechanisms of nodule N2 fixation are now well documented, some uncertainty remains as to whether N2 fixation is strictly coupled with photosynthetic carbon fixation. That is, the metabolic origin and redistribution of carbon skeletons used to incorporate nitrogen are still relatively undefined. Here, we conducted isotopic labelling with both 15N2 and 13C-depleted CO2 on alfalfa plants grown under controlled conditions and took advantage of isotope ratio mass spectrometry to investigate the relationship between carbon and nitrogen turn-over in respired CO2, total organic matter and amino acids. Our results indicate that CO2 evolved by respiration had an isotopic composition similar to that in organic matter regardless of the organ considered, suggesting that the turn-over of respiratory pools strictly followed photosynthetic input. However, carbon turn-over was nearly three times greater than N turn-over in total organic matter, suggesting that new organic material synthesised was less N-rich than pre-existing organic material (due to progressive nitrogen elemental dilution) or that N remobilisation occurred to sustain growth. This pattern was not consistent with the total commitment into free amino acids where the input of new C and N appeared to be stoichiometric. The labelling pattern in Asn was complex, with contrasted C and N commitments in different organs, suggesting that neosynthesis and redistribution of new Asn molecules required metabolic remobilisation. We conclude that the production of new organic material during alfalfa growth depends on both C and N remobilisation in different organs. At the plant level, this remobilisation is complicated by allocation and metabolism in the different organs. Additional keywords: carbon exchange, carbon isotopes, nitrogen fixation, nitrogen 15 isotope

Efficacy of the fluorescent dyes Fast Blue, Fluoro-Gold, and Diamidino Yellow for retrograde tracing to dorsal root ganglia after subcutaneous injection

The present study was designed to investigate the efficacy of the fluorescent dyes Fast Blue (FB), Fluoro-Gold (FG), and Diamidino Yellow (DY) for retrograde tracing of lumbar dorsal root ganglia after their subcutaneous injection into different hindlimb digits. Injection of equal volumes (0.5 mu l) of 5% FB or 2% FG resulted in similar mean numbers of sensory neurones labelled by each tracer. Injection of equal volumes (0.5 mu l) of FB or FG in a single digit followed 10 days later by a second injection of the same volume of 5% DY into the same digit resulted in similar mean numbers of labelled sensory neurones for each of the three tracers. Furthermore, on average, 75% of all the FB-labelled cells and 74% of all FC-labelled cells also contained DY. Repeating the same experiment with an increased volume of DY (1.5 mu l) resulted in an increase in the mean number of double-labelled profiles to 82 and 84% for FB and FG, respectively. The results show that FB, FG and DY label similar numbers of cutaneous afferents and that a high level of double labelling may be obtained after sequential injections in digits. These properties make them suitable candidates in investigations where a combination of tracers with similar labelling efficacies is needed.

Persistence of tracer in the application site -a potential confounding factor in nerve regeneration studies

Selective reinnervation of peripheral targets after nerve injury might be assessed by injecting a first tracer in a target before nerve injury to label the original neuronal population, and applying a second tracer after the regeneration period to label the regenerated population. However, altered uptake of tracer, fading, and cell death may interfere with the results. Furthermore, if the first tracer injected remains in the target tissue, available for 're-uptake' by misdirected regenerating axons, which originally innervated another region, then the identification of the original population would be confused. With the aim of studying this problem, the sciatic nerve of adult rats was sectioned and sutured. After 3 days, to allow the distal axon to degenerate avoiding immediate retrograde transport, one of the dyes: Fast Blue (FB), Fluoro-Gold (FG) or Diamidino Yellow (DY), was injected into the tibial branch of the sciatic nerve, or in the skin of one of the denervated digits. Rats survived 2-3 months. The results showed labelled dorsal root ganglion (DRG) cells and motoneurones, indicating that late re-uptake of a first tracer occurs. This phenomenon must be considered when the model of sequential labelling is used for studying the accuracy of peripheral reinnervation.

Sertifioidun puuraaka-aineen ja sertifioitujen tuotteiden tietojärjestelmän määrittely

Asiakkaiden kiinnostus tuotteiden raaka-aineiden alkuperää kohden on kasvanut. Puuraaka-aineesta tehtyjen tuotteiden alkuperä voidaan todentaa sertifikaatilla, jos puu on peräisin sertifioidusta metsästä ja sen alkuperäketju tunnetaan. Tieto sertifioidusta tuotteesta kulkeutuu asiakkaalle leimaamalla tuotteita sertifioinnista kertovin merkein tai merkitsemällä esimerkiksi tuotteen asiakirjaan sertifioinnista kertova teksti. Tutkimus tehtiin Stora Enson Imatran tehtaalle, jolla oli tarve uudelle tietojärjestelmälle. Tutkimuksen tarkoituksena oli määritellä sertifioidun puuraaka-aineen käytön ja sertifioituina myytävien tuotteiden seurannan mandollistava tietojärjestelmä. Puun alkuperäketjun hallinnan standardit määiittävät, paljonko sertifioitua puuta kohdennettava tuotteelle, jos tuotteita halutaan myydä sertifioituna. Tietojärjestelmän määrittelyn lisäksi oli tarve selvittää prosessi, kuinka tuotteisiin ja asiakirjoihin saataisiin sertifioinnista kertova leima. Tutkimusta tehdessä selvisi, että tehtaan nykyiset myyntitoiminnot ja tietojärjestelmät asettavat rajoituksia tuotteiden ja asiakirjojen leimaukselle. Prosessia kartoitettiin mm. haastatt elemalla tehtaan ti etojärj estelmi en, myynti- ja varastotoimintojen parissa työskenteleviä ihmisiä. Haastattelujen perusteella koottiin myös tietojärjestelmän asiakasvaatimukset. Vaatimusmäärittelyä tehdessä huomioitiin myös alkuperäketjun hallinnan standardien vaatimukset. Määriteltävän tietojärjestelmän toimintaa pyrittiin havainnollistamaan tehtaan edustajille prototyyppien avulla. Lisäksi tutkimuksessa on pyritty havainnollistamaan järjestelmän toimintaa erilaisin kaavioin ja kuvin. Haasteena työssä oli tulkita standardeja ja kerätä yhteen toisistaan poikkeavia asiakasvaatimuksia.

Correlative microscopy.

In recent years correlative microscopy, combining the power and advantages of different imaging system, e.g., light, electrons, X-ray, NMR, etc., has become an important tool for biomedical research. Among all the possible combinations of techniques, light and electron microscopy, have made an especially big step forward and are being implemented in more and more research labs. Electron microscopy profits from the high spatial resolution, the direct recognition of the cellular ultrastructure and identification of the organelles. It, however, has two severe limitations: the restricted field of view and the fact that no live imaging can be done. On the other hand light microscopy has the advantage of live imaging, following a fluorescently tagged molecule in real time and at lower magnifications the large field of view facilitates the identification and location of sparse individual cells in a large context, e.g., tissue. The combination of these two imaging techniques appears to be a valuable approach to dissect biological events at a submicrometer level. Light microscopy can be used to follow a labelled protein of interest, or a visible organelle such as mitochondria, in time, then the sample is fixed and the exactly same region is investigated by electron microscopy. The time resolution is dependent on the speed of penetration and fixation when chemical fixatives are used and on the reaction time of the operator for cryo-fixation. Light microscopy can also be used to identify cells of interest, e.g., a special cell type in tissue or cells that have been modified by either transfections or RNAi, in a large population of non-modified cells. A further application is to find fluorescence labels in cells on a large section to reduce searching time in the electron microscope. Multiple fluorescence labelling of a series of sections can be correlated with the ultrastructure of the individual sections to get 3D information of the distribution of the marked proteins: array tomography. More and more efforts are put in either converting a fluorescence label into an electron dense product or preserving the fluorescence throughout preparation for the electron microscopy. Here, we will review successful protocols and where possible try to extract common features to better understand the importance of the individual steps in the preparation. Further the new instruments and software, intended to ease correlative light and electron microscopy, are discussed. Last but not least we will detail the approach we have chosen for correlative microscopy.

Characterisation of anatomical properties of the human basal ganglia using magnetic resonance imaging

The detailed in-vivo characterization of subcortical brain structures is essential not only to understand the basic organizational principles of the healthy brain but also for the study of the involvement of the basal ganglia in brain disorders. The particular tissue properties of basal ganglia - most importantly their high iron content, strongly affect the contrast of magnetic resonance imaging (MRI) images, hampering the accurate automated assessment of these regions. This technical challenge explains the substantial controversy in the literature about the magnitude, directionality and neurobiological interpretation of basal ganglia structural changes estimated from MRI and computational anatomy techniques. My scientific project addresses the pertinent need for accurate automated delineation of basal ganglia using two complementary strategies: ? Empirical testing of the utility of novel imaging protocols to provide superior contrast in the basal ganglia and to quantify brain tissue properties; ? Improvement of the algorithms for the reliable automated detection of basal ganglia and thalamus Previous research demonstrated that MRI protocols based on magnetization transfer (MT) saturation maps provide optimal grey-white matter contrast in subcortical structures compared with the widely used Tl-weighted (Tlw) images (Helms et al., 2009). Under the assumption of a direct impact of brain tissue properties on MR contrast my first study addressed the question of the mechanisms underlying the regional specificities effect of the basal ganglia. I used established whole-brain voxel-based methods to test for grey matter volume differences between MT and Tlw imaging protocols with an emphasis on subcortical structures. I applied a regression model to explain the observed grey matter differences from the regionally specific impact of brain tissue properties on the MR contrast. The results of my first project prompted further methodological developments to create adequate priors for the basal ganglia and thalamus allowing optimal automated delineation of these structures in a probabilistic tissue classification framework. I established a standardized workflow for manual labelling of the basal ganglia, thalamus and cerebellar dentate to create new tissue probability maps from quantitative MR maps featuring optimal grey-white matter contrast in subcortical areas. The validation step of the new tissue priors included a comparison of the classification performance with the existing probability maps. In my third project I continued investigating the factors impacting automated brain tissue classification that result in interpretational shortcomings when using Tlw MRI data in the framework of computational anatomy. While the intensity in Tlw images is predominantly

Chlamydiaceae and Chlamydia-like organisms in the koala (Phascolarctos cinereus)--organ distribution and histopathological findings.

Chlamydial infections in koalas can cause life-threatening diseases leading to blindness and sterility. However, little is known about the systemic spread of chlamydiae in the inner organs of the koala, and data concerning related pathological organ lesions are limited. The aim of this study was to perform a thorough investigation of organs from 23 koalas and to correlate their histopathological lesions to molecular chlamydial detection. To reach this goal, 246 formalin-fixed and paraffin embedded organ samples from 23 koalas were investigated by histopathology, Chlamydiaceae real-time PCR and immunohistochemistry, ArrayTube Microarray for Chlamydiaceae species identification as well as Chlamydiales real-time PCR and sequencing. By PCR, two koalas were positive for Chlamydia pecorum whereas immunohistochemical labelling for Chlamydiaceae was detected in 10 tissues out of nine koalas. The majority of these (n=6) had positive labelling in the urogenital tract related to histopathological lesions such as cystitis, endometritis, pyelonephritis and prostatitis. Somehow unexpected was the positive labelling in the gastrointestinal tract including the cloaca as well as in lung and spleen indicating systemic spread of infection. Uncultured Chlamydiales were detected in several organs of seven koalas by PCR, and four of these suffered from plasmacytic enteritis of unknown aetiology. Whether the finding of Chlamydia-like organisms in the gastrointestinal tract is linked to plasmacytic enteritis is unclear and remains speculative. However, as recently shown in a mouse model, the gastrointestinal tract might play a role being the site for persistent chlamydial infections and being a source for reinfection of the genital tract.

Facilitar la teoría de la mente en niños con alteraciones del lenguaje

The ability to recognize mental states in oneself and others, or the theory of mind, has been a widely studied topic in last years. Traditionally, these studies have focused on the understanding of false belief, one of the most important milestones in its development. Studies conducted with children with language difficulties suggest that they have a delay in false belief acquisition. On the other hand, numerous studies have established a relationship between the understanding of false belief and language skills. In particular, it has been suggested that training based in labelling objects with double perspective facilitates the understanding of false belief. The aim of the present study is to test the effect of the structuration level of labelling training in a group of children with difficulties in language development. A total of 14 children, aged between 58 and 107 months, were trained in 3 sessions, and a pre-test and a post-test were administered to evaluate the effects of the training. The results confirm a delay in false belief understanding in children with developmental language difficulties, and suggest that the structured labelling training is effective to improve this understanding

Aplicação da espectroscopia de ressonância magnética nuclear para estudos de difusão molecular em líquidos: a técnica DOSY

Diffusion coefficients provide uniquely detailed and easily interpreted information on molecular organization and phase structure. They are quite sensitive to structural changes, and to binding and association phenomena, in particular for liquid colloidal or macromolecular systems. This paper describes the principles of diffusion measurements in liquids by pulsed magnetic field gradient spin-echo (PFG-SE) NMR spectroscopy. The important PFG-SE technique known as DOSY is presented and discussed. This is a noninvasive technique that can provide individual multicomponent translational diffusion coefficients with good precision in a few minutes, without the need for radioactive isotopic labelling.

In vivo co-ordinated interactions between inhibitory systems to control glutamate mediated hippocampal excitability.

We present an overview of the long-term adaptation of hippocampal neurotransmission to cholinergic and GABAergic deafferentation caused by excitotoxic lesion of the medial septum. Two months after septal microinjection of 2.7 nmol a -amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA), a 220% increase of GABA A receptor labelling in the hippo- campal CA3 and the hilus was shown, and also changes in hippocampal neurotransmission characterised by in vivo microdialysis and HPLC. Basal amino acid and purine extra- cellular levels were studied in control and lesioned rats. In vivo effects of 100 m M KCl perfusion and adenosine A 1 receptor blockade with 1,3-dipropyl- 8-cyclopentylxanthine (DPCPX) on their release were also investigated. In lesioned animals GABA, glutamate and glutamine basal levels were decreased and taurine, adenosine and uric acid levels increased. A similar response to KCl infusion occurred in both groups except for GABA and glutamate, which release decreased in lesioned rats. Only in lesioned rats, DPCPX increased GABA basal level and KCl-induced glutamate release, and decreased glutamate turnover. Our results evidence that an excitotoxic septal lesion leads to increased hippocampal GABA A receptors and decreased glutamate neurotransmis- sion. In this situation, a co-ordinated response of hippocampal retaliatory systems takes place to control neuron excitability.

Veus ignorades: representacions socials, subjectivitat i resiliència en persones grans amb pèrdua d’autonomia funcional

Els processos per a l’aprovació i la implementació de la Llei per a l’Autonomia personal i l’Atenció a les Persones en situació de Dependència (LAPAD) han donat lloc a un intens debat polític i social que, coincidint també amb les millores en la provisió de serveis i els avenços mèdics, ha contribuït a un procés de classificació i d’etiquetatge basats en els dèficits de les persones que es troben en aquestes circumstàncies. Aquesta visió anul·la el subjecte i la seva experiència singular i condiciona l’abordatge dels models d’atenció i de cura. L’estudi pretén fer una aproximació a les persones grans amb pèrdua d’autonomia funcional, fent emergir les seves veus, que expressen com perceben, interpreten, afronten i es reajusten a la nova situació. Partint d’un enfocament constructivista, basat en la subjectivitat, es fa un recorregut sobre els models de la discapacitat que han reeixit en l’activitat científica dels darrers anys, els mecanismes de regulació de les pèrdues que defensen les teories del cicle vital i les aportacions que s’han fet sobre el model de la resiliència aplicat a les persones que envelleixen. El resultats de l’estudi mostren com les representacions i els significats que les persones grans atribueixen a la seva experiència s’inscriuen en les seves trajectòries vitals, donant un sentit únic i singular a la forma de viure i de respondre a la pèrdua d’autonomia funcional i les seves conseqüències. Aquelles que expressen una vivència d’integritat respecte de la vida viscuda, amb predomini d’afectes positius envers un mateix i els altres, que conserven l’esperança i el desig de continuar vivint, s’ajusten a les pèrdues de manera més satisfactòria que aquelles que expressen desconfiança i una certa amargor respecte de la pròpia vida. D’això se’n deriva que els espais d’escolta i d’acompanyament poden ser un recurs vàlid i necessari en el qual, a través de la paraula i el testimoni narrat, el subjecte pugui repensar i resignificar les seves experiències.

Rhizodeposition of organic carbon by plants with contrasting traits for resource acquisition: responses to different fertility regimes

Background and aims Rhizodeposition plays an important role in mediating soil nutrient availability in ecosystems. However, owing to methodological difficulties (i.e., narrow zone of soil around roots, rapid assimilation by soil microbes) fertility-induced changes in rhizodeposition remain mostly unknown. Methods We developed a novel long-term continuous 13C labelling method to address the effects of two levels of nitrogen (N) fertilization on rhizodeposited carbon (C) by species with different nutrient acquisition strategies. Results Fertility-induced changes in rhizodeposition were modulated by root responses to N availability rather than by changes in soil microbial biomass. Differences among species were mostly related to plant biomass: species with higher total leaf and root biomass also had higher total rhizodeposited C, whereas species with lower root biomass had higher specific rhizodeposited C (per gram root mass). Experimental controls demonstrated that most of the biases commonly associated with this type of experiment (i.e., long-term steady-state labelling) were avoided using our methodological approach. Conclusions These results suggest that the amount of rhizodeposited C from plants grown under different levels of N were driven mainly by plant biomass and root morphology rather than microbial biomass. They also underline the importance of plant characteristics (i.e., biomass allocation) as opposed to traits associated with plant resource acquisition strategies in predicting total C rhizodeposition.

The Child Protection System from the Perspective of Young People: Messages from 3 Studies

This article reports findings and reflections based on the results of three different research projects conducted between 2008 and 2013 and focusing on the perspective of young care leavers in Spain. The overall aim was to examine these young people’s perceptions and evaluations of how they were treated while in the public care system, mainly residential care. Reviewing these qualitative studies, the most common and relevant issues highlighted by young people were related to the following themes: (a) entering care; (b) stability and emotional bonds in care; (c) education; (d) friends; (e) labelling, stigmatization, rights and opportunities; (f) autonomy and responsibility versus overprotection; (g) contact with parents, siblings and extended family; (h) maltreatment in care; and (i) leaving care. One of the main elements used in their assessments was comparison (i) between their previous situation within their birth family and the quality of care experienced in the residential home; and (ii) between what these young people commonly refer to as “normal children” and children in care. Recommendations deriving from their advice and opinions are also debated

O sistema globalmente harmonizado de classificação e rotulagem de produtos químicos (GHS): uma introdução para sua aplicação em laboratórios de ensino e pesquisa acadêmica

The present article provides an overview of the Globally Harmonised System of Classification, Labelling and Packaging of Chemicals (GHS) and its implementation in Brazil. Although Classification and Packaging is beyond the scope of the responsibility of academic chemists, labelling of chemicals used in academic laboratories will be required by the competent authorities to ensure the safety of students and staff. Therefore, academic teachers and researchers responsible should be familiarised with the GHS principles outlined here and at least be able to label, by applying these principles, mixtures of substances previously classified by the competent authorities.

Produtos químicos perigosos utilizados em laboratórios de ensino: proposta e exemplos para indicação de seus perigos no rótulo

Typically hundreds of different chemical products stored in small flasks which cannot accommodate labels large enough to bear all information required by the Globally Harmonised System of Classification, Labelling and Packaging of Chemicals (GHS) are used in academic laboratories. To overcome this restriction Brazilian legislation permits communication of the dangers of chemical products to trained laboratory workers/students by alternative means. Here we describe an alternative label system, developed by the German Social Accident Insurance (DGUV), which allows hazard communication to trained workers/students via a labelling system, using pictograms and highly condensed hazard statements to alert about dangerous chemical products.