Impaired expression of the plastidic ferrochelatase by antisense RNA synthesis leads to a necrotic phenotype of transformed tobacco plants

Protoporphyrin IX is the last common intermediate of tetrapyrrole biosynthesis. The chelation of a Mg2+ ion by magnesium chelatase and of a ferrous ion by ferrochelatase directs protoporphyrin IX towards the formation of chlorophyll and heme, respectively. A full length cDNA clone encoding a ferrochelatase was identified from a Nicotiana tabacum cDNA library. The encoded protein consists of 497 amino acid residues with a molecular weight of 55.4 kDa. In vitro import of the protein into chloroplasts and its location in stroma and thylakoids confirm its close relationship to the previously described Arabidopsis thaliana plastid-located ferrochelatase (FeChII). A 1700-bp tobacco FeCh cDNA sequence was expressed in Nicotiana tabacum cv. Samsun NN under the control of the CaMV 35S promoter in antisense orientation allowing investigation into the consequences of selective reduction of the plastidic ferrochelatase activity for protoporphyrin IX channeling in chloroplasts and for interactions between plastidic and mitochondrial heme synthesis. Leaves of several transformants showed a reduced chlorophyll content and, during development, a light intensity-dependent formation of necrotic leaf lesions. In comparison with wild-type plants the total ferrochelatase activity was decreased in transgenic lines leading to an accumulation of photosensitizing protoporphyrin IX. Ferrochelatase activity was reduced only in plastids but not in mitochondria of transgenic plants. By means of the specifically diminished ferrochelatase activity consequences of the selective inhibition of protoheme formation for the intracellular supply of heme can be investigated in the future.     

Expression and secretion of ficolin beta by porcine neutrophils

Ficolins are collagenous lectins that bind N-acetylglucosamine (GlcNAc) as well as some bacterial surfaces, and may have opsonic and complement-activating functions. Ficolin alpha in porcine plasma binds Actinobacillus pleuropneumoniae serotype 5 (APP) in a GlcNAc-dependent manner. In the present study, we discovered that porcine neutrophils, but not platelets or mononuclear cells, contained a different ficolin that migrated as a 39-kDa band on SDS-PAGE and resembled a minor component of plasma ficolins that binds APP. However, neutrophil ficolins (pI range 6.4-7.4) were readily distinguished from plasma ficolin alpha (pI 5.2-5.8) by 2D PAGE. Neutrophil ficolin was consistent with ficolin beta by pI and peptide mass fingerprinting with matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Porcine neutrophils expressed ficolin beta, but not ficolin alpha, as determined by RT-PCR. Ficolin beta was present in the membrane and cytoplasmic fractions of nonactivated neutrophils, but the majority of ficolin beta was secreted upon activation with PMA. Ficolin alpha readily bound to intact APP, but ficolin beta did not under the same conditions. These studies demonstrate that neutrophils express ficolin beta and secrete it when activated; however, ficolin beta may have different binding functions than ficolin alpha.     

Specific Expression of N-Acetylaspartate in Neurons, Oligodendrocyte-Type-2 Astrocyte Progenitors, and Immature Oligodendrocytes In Vitro

To test the specificity of N-acetylaspartate (NAA) as a neuronal marker for proton nuclear magnetic resonance (1H NMR) spectroscopy, purified and characterized cultured cells were analyzed for their NAA content using both 1H NMR and HPLC. Cell types studied included cerebellar granule neurons, type-1 astrocytes, meningeal cells, oligodendrocyte-type-2 astrocyte (O-2A) progenitor cells, and oligodendrocytes. A high concentration of NAA was found in extracts of cerebellar granule neurons (approximately 12 nmol/mg of protein), whereas NAA remained undetectable in purified type-1 astrocytes, meningeal cells, and mature oligodendrocytes. However, twice the neuronal level of NAA was found in O-2A progenitors grown in vitro. In addition significant levels of NAA were also detected in cultures of immature oligodendrocytes. Our data partly support previous suggestions that NAA may be a useful neuronal marker for 1H NMR spectroscopic examination of the adult brain. However, they also raise the further possibility that alterations of NAA associated with some specific brain disorders, particularly disorders seen in newborn and young children, may reflect abnormalities in the development of oligodendroglia or their precursors.     

Changes in blood glutathione concentrations, and in erythrocyte glutathione reductase and glutathione S-transferase activity after running training and after participation in contests

Previously sedentary men (n = 23) and women (n = 18) were trained to run a half marathon contest after 40 weeks. Total blood glutathione had increased by 20 weeks of training and had returned to normal after 40 weeks. Erythrocyte glutathione reductase activity had increased by 20 weeks and remained elevated after 40 weeks. This effect was accompanied by decreases in glutathione reductase coefficients, which indicated that increases in the presence of riboflavin may have been responsible for the changes in reductase activity. Erythrocyte glutathione S-transferase activity had increased slightly after 20 weeks of training and a much more marked increase was found after 40 weeks. This may have been indicative of the occurrence of lipid peroxidation in this phase of training. The participants ran a 15-km race after the first 20 weeks of training and a half marathon after 40 weeks. Blood glutathione tended to decrease after the 15-km race and increased after the half marathon. In both cases it had returned to normal values 5 days after the race. Erythrocyte glutathione reductase was elevated 1 day after the races, and had returned to normal after 5 days. This could also have been explained from concurrent changes in the riboflavin content of the erythrocytes. Erythrocyte glutathione S-transferase activity decreased after both races, but was restored 5 days after the half marathon while such was not the case after the 15-km race.     

Life cycle inventory dataset review criteria—a new proposal

Purpose

A review of LCA process datasets is an important element of quality assurance for databases and for other systems to provide LCA datasets. Somewhat surprisingly, a broadly accepted and applicable set of criteria for a review of LCA process datasets was lacking so far. Different LCA databases and frameworks are proposing and using different criteria for reviewing datasets. To close this gap, a set of criteria for reviewing LCA dataset has been developed within the Life Cycle Initiative.

Methods

Previous contributions to LCA dataset review have been analysed for a start, from ISO and various LCA databases. To avoid somewhat arbitrary review criteria, four basic rules are proposed which are to be fulfilled by any dataset. Further, concepts for assessing representativeness and relevance are introduced into the criteria set from established practices in statistics and materiality. To better structure the criteria and to ease their application, they are grouped into clusters. A first version of the developed review criteria was presented in two workshops with database providers and users on different levels of experience, and draft versions of the criteria were shared within the initiative. The current version of the criteria reflects feedback received from various stakeholders and has been applied and tested in a review for newly developed datasets in Brazil, Malaysia and Thailand.

Results and discussion

Overall, 14 criteria are proposed, which are organised in clusters. The clusters are goal, model, value, relevance and procedure. For several criteria, a more science-based definition and evaluation is proposed in comparison to ‘traditional’ LCA. While most of the criteria depend on the goal and scope of dataset development, a core set of criteria are seen as essential and independent from specific LCA modelling. For all the criteria, value scales are developed, typically using an ordinal scale, following the pedigree approach.

Conclusions

Review criteria for LCI datasets are now defined based on a stringent approach. They aim to be globally acceptable, considering also database interoperability and database management aspects, as well as feedback received from various stakeholders, and thus close an important gap in LCA dataset quality assurance. The criteria take many elements of already existing criteria but are the first to fully reflect the implications of the ISO data quality definition, and add new concepts for representativeness and relevance with the idea to better reflect scientific practice outside of the LCA domain. A first application in a review showed to be feasible, with a level of effort similar to applying other review criteria. Aspects not addressed yet are the review procedure and the mutual recognition of dataset reviews, and their application for a very high number of datasets.

     

Intensification of agriculture in southwestern Germany between the Bronze Age and Medieval period,based on archaeobotanical data from Baden-Württemberg

Vegetation History and Archaeobotany - A system of farming with an alternation of land use between being cultivated or left fallow as grassland (Feldgraswirtschaft) developed in southwestern...     

You Do the Math: Coding of Bets and Outcomes in a Gambling Task in the Feedback-Related Negativity and P300 in Healthy Adults

The feedback-related negativity (FRN) is an event-related potential (ERP) component associated with processing of performance feedback, with more negative amplitudes for losses relative to wins. The amplitude of the FRN following near misses, i.e. the experience of coming close to winning, is between the amplitude elicited by losses and wins. In gambling, however, outcome value may not always be obvious since initially placed bets need to be taken into account when evaluating wins or losses. It is still unclear if initial bet size is reflected in the FRN or the later P300 component. The present study applied a virtual card gambling task to investigate the sensitivity of FRN and P300 to the manipulation of outcome magnitude as implemented through the presence or absence of initial bets, resulting in wins, losses or ambivalent outcomes, with the latter representing losses with and wins without bets. The FRN was larger for trials with bets compared to trials without bets. Wins were associated with a smaller FRN than losses or ambivalent outcomes, while losses and ambivalent outcomes did not differ. P300 amplitudes were larger for trials without bets, and wins were associated with a larger P300 than losses or ambivalent outcomes. Crucially, P300 amplitudes were also smaller for ambivalent outcomes compared to losses. Thus, the different dimensions determining outcome value appear to be integrated in early and late stages of feedback processing. However, only at later stages reflected in the P300 were ambivalent outcomes with and without bet clearly distinguished from other outcomes.     

Emotion recognition in children and adolescents with attention-deficit/hyperactivity disorder (ADHD)

Children with attention-deficit/hyperactivity disorder (ADHD) are impaired in social adaptation and display deficits in social competence. Deficient emotion recognition has been discussed to underlie these social problems. However, comorbid conduct problems have not been considered in the majority of studies conducted so far, and the influence of medication on emotion recognition has rarely been studied. Here, emotion recognition performance was assessed in children with ADHD without medication compared with children with ADHD under stimulant medication and a matched control group. In order to rule out confounding by externalizing symptoms, children with comorbid conduct problems were excluded. Video clips with neutral faces developing a basic emotion (happiness, sadness, disgust, fear and anger) were presented in order to assess emotion recognition. Results indicated between-group differences neither concerning the number of correctly identified emotions nor concerning reaction times and their standard deviations. Thus, we suggest that ADHD per se is not associated with deficits in emotion recognition.     

Seminal roots of wild and cultivated barley differentially respond to osmotic stress in gene expression,suberization, and hydraulic conductivity

Wild barley, Hordeum vulgare spp. spontaneum, has a wider genetic diversity than its cultivated progeny, Hordeum vulgare spp. vulgare. Osmotic stress leads to a series of different responses in wild barley seminal roots, ranging from no changes in suberization to enhanced endodermal suberization of certain zones and the formation of a suberized exodermis, which was not observed in the modern cultivars studied so far. Further, as a response to osmotic stress, the hydraulic conductivity of roots was not affected in wild barley, but it was 2.5-fold reduced in cultivated barley. In both subspecies, osmotic adjustment by increasing proline concentration and decreasing osmotic potential in roots was observed. RNA-sequencing indicated that the regulation of suberin biosynthesis and water transport via aquaporins were different between wild and cultivated barley. These results indicate that wild barley uses different strategies to cope with osmotic stress compared with cultivated barley. Thus, it seems that wild barley is better adapted to cope with osmotic stress by maintaining a significantly higher hydraulic conductivity of roots during water deficit.     

A UNEP/SETAC approach towards a life cycle sustainability assessment—our contribution to Rio+20

Purpose

To contribute to the upcoming United Nations Conference on Sustainable Development (Rio+20) in 2012 by introducing a life cycle sustainability assessment (LCSA) and showing how it can play a crucial role in moving towards sustainable consumption and production. The publication, titled Towards a Life Cycle Sustainability Assessment, and published by the UNEP/SETAC Life Cycle Initiative aims to show how three life cycle techniques—(environmental) LCA, S-LCA and LCC—can be combined as part of an over-arching LCSA.

Methods

The method was demonstrated by evaluating the characteristics of each phase for each life cycle technique. In defining the goal and scope of an LCSA, for example, different aspects should be taken into account to establish the aim of the study as well as the functional unit, system boundaries, impact category and allocation. Then, the data to be collected for the life cycle sustainability inventory can be either in a unit process or on an organisational level. They can also be quantitative or qualitative. Life cycle sustainability impact assessment should consider the relevance of the impacts as well as the perspective of stakeholders. The interpretation should not add up the results, but rather evaluate them jointly. In order to clarify the approach, a case study is presented to evaluate three types of marble according to the proposed method.

Results and discussion

The authors have identified that while LCSA is feasible, following areas need more development: data production and acquisition, methodological development, discussion about LCSA criteria (e.g. cutoff rules), definitions and formats of communication and dissemination of LCSA results and the expansion of research and applications combining (environmental) LCA, LCC and S-LCA. The authors also indicate that it is necessary to develop more examples and cases to improve user capacity to analyse the larger picture and therefore address the three dimensions or pillars of sustainability in a systematic way. Software and database providers are called for in order to facilitate user-friendly and accessible tools to promote LCSAs.

Conclusions

The application demonstrated that, although methodological improvements are still needed, important steps towards an overarching sustainability assessment have been accomplished. LCSA is possible and should be pursued; however, more efforts should be made to improve the technique and facilitate the studies in order to contribute to a greener economy.