In vitro culture and medium-term conservation of the rare wild species Gladiolus imbricatus

Gladiolus imbricatus, a rare species spread in the hill region of Europe, is resistant to abiotic and biotic stress being one of the most cold-tolerant in the genus. Moreover it contains high vitamin C and minerals in the leaves and the flowers are considered as edible. The aim of our study was to develop in vitro technologies for micropropagation, multiplication, corm development, somatic embryogenesis and medium-term storage of this endangered species, Red listed in South-Eastern and Central European countries. Initiation of in vitro cultures was started from wounded seeds or excised mature embryos. Micropropagation and multiplication was achieved on Murashige Skoog (MS) solid medium with 30 g L-1 sucrose and 1 mg L-1 N-6-benzyl adenine (BA). The callus developed from wounded seeds on a richer MS medium proved to be embryogenic. Increased concentrations of sucrose promote corm development as also shown for other genotypes of cultivated gladioli. Further on a two-step culture protocol, on solid and liquid shaken MS based media, each for 6 months was done to evaluate the effects of acetic acid and ancymidol on corm development. Acetic acid + sucrose 9%, alone or in combination with ancymidol stimulated corm formation and yield. For medium term conservation maintenance of the in vitro cultures in low temperature in the dark proved to be the best in reducing the growth rate of the shoots after 3, 6 and 12 months. The recovery of plant growth was evaluated after 12 months, by the transfer to normal growth conditions. 25% of the plants were recovered after one year storage. Besides the importance for biotechnology, the in vitro techniques described here might be used, after molecular analysis of genetic stability, for the restoration of natural populations into the habitats where this species became extinct.Keywords: Acetic acid, ancymidol, corm development, somatic embryogenesi

Alcohol use in the first three years of bereavement: a national representative survey

BACKGROUND:Earlier results concerning alcohol consumption of bereaved persons are contradictory. The aim of the present study was to analyze the relationship between bereavement and alcohol consumption accounting for time and gender differences on a nationally representative sample from Hungary ("Hungarostudy Epidemiological Panel Survey", N = 4457)METHODS:Drinking characteristics of mourning persons (alcohol consumption, dependence symptoms, and harmful consequences of alcohol use) in the first three years of grief were examined among persons between 18-75 years using the Alcohol Use Disorders Identification Test (AUDIT).RESULTS:Men bereaved for one year scored higher on two dimensions of AUDIT (dependence symptoms and harmful alcohol use), while men bereaved for two years scored higher on all three dimensions of AUDIT compared to the non-bereaved. The rate of men clinically at-risk concerning alcohol consumption among the non-bereaved is 12.9%, and among men bereaved for one year is 18.4% (a non-significant difference), while 29.8% (p<0.001, OR=2,781) among men bereaved for two years. However, men bereaved for three years did not differ from the non-bereaved in their drinking habits. In case of bereaved women, again no difference was found with respect to alcohol use compared to the non-bereaved.CONCLUSION:Among bereaved men, the risk of alcohol related problems tends to be higher, which can be shown both among men bereaved for one year as well as men bereaved for two years. Considering the higher morbidity and mortality rates of bereaved men, alcohol consumption might play a mediator role. These facts draw attention to the importance of prevention, early recognition, and effective therapy of hazardous drinking in bereaved men

Bond behaviour of twisted stainless steel bars in mortar joints

The use of twisted stainless steel bars has proven to be effective in scenarios where lateral loads (e. g. earthquakes and winds storms) can cause partial or complete out-of-plane collapse of masonry wall’s outer leaves or separation of wood diaphragms from masonry walls. The particular application of these bars as a dry system in mortar joints, without any binder, brings additional advantages in terms of cost, installation time and weather restrictions. An experimental campaign composed by 60 pull-out tests aimed at characterizing the bond behaviour of twisted stainless steel bars in mortar joints. The influence on bond behaviour of two diameters of the helibar (ϕ8 mm and ϕ10 mm), three different anchorage lengths (8ϕhb, 12ϕhb, and 20ϕhb), and two diameters of the pre-drilled holes (ϕhb–2 mm and ϕhb–4 mm) was studied. Bond strength increased for the tighter pre-drilled hole but decreased for the higher twisted steel bar diameter. Good correlations between bond strength and anchorage length were found for the less tight pre-drilled holes

Flexural out-of-plane retroftting technique for masonry walls in historical constructions

The paper presents a recent technology for the out-of-plane flexural strengthening of historic masonry walls. The method is based on the forming of an irregular net following the joint texture of the wall, which would be able to work in tension. The applied net can be hidden by repointing the joints. This way the technology does not cause visual alteration to walls without rendering, respecting their authenticity. An ongoing research in Portugal is aiming to increase the workability and efficiency of the technique by developing a new anchor element for the application of the net. The discussion includes the introduction of the method and partial results of the research from the University of Minho, Portugal

Contribution à l'évaluation de l'impact environnemental des bâtiments (modélisation numérique des émissions dans l'eau des substances dangereuses)

Tous les produits de construction ou les ouvrages qui sont en contact avec l eau de pluie, sont susceptibles de relarguer des composants chimiques potentiellement dangereux pour la santé humaine et l environnement. Actuellement, le relargage des substances dangereuses dans l eau est évalué essentiellement par des méthodes expérimentales - généralement des essais de lixiviation réalisées à l échelle du laboratoire. Par ailleurs, les impacts environnementaux des produits de construction et des bâtiments sont évalués par l approche Analyse de Cycle de Vie (ACV), réalisées à partir de données propres aux fabricants et de données génériques existantes constituées en bases de données. Or, les émissions de polluants pendant la vie en oeuvre des produits de construction sont actuellement mal prises en compte dans ces mêmes bases de données existantes et dans les méthodes utilisées pour l évaluation des performances environnementales des produits et des bâtiments.Dans ce contexte, les objectifs de ces travaux ont été : (i) de développer une méthodologie pour l évaluation du comportement à la lixiviation de différentes matrices de produits de construction et (ii) d intégrer les données de lixiviation dans les méthodes ACV à l échelle du bâtiment. La méthodologie développée pour l évaluation du comportement à la lixiviation des produits de construction est basée sur : (1) des essais de caractérisation à la lixiviation, selon les méthodes standardisées d évaluation répondant au Règlement Produits de Construction (RPC), (2) la modélisation et la simulation des phénomènes couplés chimie-transport à l échelle laboratoire et vraie grandeur et (3) l intégration des données de lixiviation, obtenues par des simulations numériques en conditions d exposition naturelle des produits de construction, dans les méthodes ACV, aux échelles produit et bâtiment.La méthodologie proposée a été appliquée et validée sur trois produits de construction : le bois traité CBA (Cuivre - Bore - Azole), la plaque fibrociment et la membrane bitumineuse. Les comportements à la lixiviation du bois traité CBA et de la plaque fibrociment a été simulé pour des conditions naturelles d exposition à long terme, en utilisant le logiciel géochimique PHREEQC®. Les modèles chimiques développées permettent de considérer les matrices organiques et minérales et leurs interactions avec les polluants. Les phénomènes de transport des substances ont été également modélisés. Les modèles d émission développés (couplage chimie-transport) permettent de simuler la lixiviation dynamique des produits considérés, pour différentes conditions d exposition. Pour la membrane bitumineuse, seule la caractérisation expérimentale a été réalisée. Les données de lixiviation obtenues lors des simulations sur la durée de vie en oeuvre des produits ont été intégrées dans les inventaires respectifs du cycle de vie (ICV) des produits. Des méthodes d impact spécifiques ont été utilisées en association avec les inventaires enrichis pour évaluer les impacts pollution de l eau , toxicité humaine et écotoxicité des produits considérés. L ACV d une maison simplifiée a été réalisée en utilisant le logiciel ELODIE. La méthode d évaluation à l échelle bâtiment consiste à intégrer les nouvelles données construites sur la base de la méthodologie développée à l échelle produit aux autres données d ACV à l échelle bâtiment. La méthodologie développée au cours de ces travaux permet donc l intégration des données de lixiviation dans les outils ACV dédiés au bâtimentAny construction product and building undergoing contact with water during its life cycle can release chemical compounds potentially hazardous for the human health and the environment. The release of dangerous substances is presently investigated especially by experimental approaches commonly by laboratory leaching tests while the environmental impacts of construction products and buildings are evaluated by a Life Cycle Assessment (LCA) approach. The dangerous substances release during use stage of construction products (leaching behaviour) is currently very poorly represented in the existing data bases and methods for environmental assessment of construction products and buildings.In this context, the main objectives of this study were: (i) to develop a methodology for the assessment of the leaching behaviour of various construction products and (ii) to integrate the leaching data in LCA approach at building scale. The methodology developed for the assessment of the leaching behaviour of construction products is based on: (1) characterisation leaching tests at lab scale, following the horizontal standardised assessment methods for harmonised approaches relating to dangerous substances under the Construction Products Regulation (CPR), (2) modelling and simulation of coupled chemical and transport phenomena at lab and field scale, and (3) utilisation of simulated leaching data for the construction products exposed in natural condition with the LCA method for the characterization of the hazardous effect during the use stage. The proposed methodology was applied and validated on three different classical construction products i.e. CBA (Copper-Boron-Azole) treated wood, fibre-cement sheets and bitumen membranes. The leaching behaviour of CBA treated wood and fibre-cement sheets was simulated over several years under natural exposure conditions, using the geochemical software PHREEQC. The chemical models consider both the mineral and the organic matrixes and their interaction with treatmentcompounds. Mass transfer and transport phenomena were modelled. The developed coupled chemical-transport models are able to represent the dynamic leaching behaviour of the respective products in various leaching conditions. For bitumen membranes only experimental characterisation was possible. The leaching data obtained by simulation over the whole use stage of the products were integrated in the life cycle inventory (LCI) and the relevant life cycle impact assessment (LCIA) methods were applied with the enriched inventory. A simplified single-family house was modelled using a software designed to evaluate the intrinsic environmental performances of a building over its entire life cycle. ELODIE software was used in this work. The building scale assessment methodology is based on coupling the methodology developed for the product scale with the Life Cycle Assessment (LCA) at building scale. This methodology allows a proper integration of leaching data in LCA tools and answers several technical questions raised in the fieldTOULOUSE-INSA-Bib. electronique (315559905) / SudocSudocFranceF

Suppression of AMPK/aak-2 by NRF2/SKN-1 down-regulates autophagy during prolonged oxidative stress.

NF-E2-related factor 2 (NRF2) transcription factor has a fundamental role in cell homeostasis maintenance as one of the master regulators of oxidative and electrophilic stress responses. Previous studies have shown that a regulatory connection exists between NRF2 and autophagy during reactive oxygen species-generated oxidative stress. The aim of the present study was to investigate how autophagy is turned off during prolonged oxidative stress, to avoid overeating and destruction of essential cellular components. AMPK is a key cellular energy sensor highly conserved in eukaryotic organisms, and it has an essential role in autophagy activation at various stress events. Here the role of human AMPK and its Caenorhabditis elegans counterpart AAK-2 was explored upon oxidative stress. We investigated the regulatory connection between NRF2 and AMPK during oxidative stress induced by tert-butyl hydroperoxide (TBHP) in HEK293T cells and C. elegans. Putative conserved NRF2/protein skinhead-1 binding sites were found in AMPK/aak-2 genes by in silico analysis and were later confirmed experimentally by using EMSA. After addition of TBHP, NRF2 and AMPK showed a quick activation; AMPK was later down-regulated, however, while NRF2 level remained high. Autophagosome formation and Unc-51-like autophagy activating kinase 1 phosphorylation were initially stimulated, but they returned to basal values after 4 h of TBHP treatment. The silencing of NRF2 resulted in a constant activation of AMPK leading to hyperactivation of autophagy during oxidative stress. We observed the same effects in C. elegans demonstrating the conservation of this self-defense mechanism to save cells from hyperactivated autophagy upon prolonged oxidative stress. We conclude that NRF2 negatively regulates autophagy through delayed down-regulation of the expression of AMPK upon prolonged oxidative stress. This regulatory connection between NRF2 and AMPK may have an important role in understanding how autophagy is regulated in chronic human morbidities characterized by oxidative stress, such as neurodegenerative diseases, certain cancer types, and in metabolic diseases.-Kosztelnik, M., Kurucz, A., Papp, D., Jones, E., Sigmond, T., Barna, J., Traka, M. H., Lorincz, T., Szarka, A., Banhegyi, G., Vellai, T., Korcsmaros, T., Kapuy, O. Suppression of AMPK/aak-2 by NRF2/SKN-1 down-regulates autophagy during prolonged oxidative stress

Customized bioreactor enables the production of 3D diaphragmatic constructs influencing matrix remodeling and fibroblast overgrowth

The production of skeletal muscle constructs useful for replacing large defects in vivo, such as in congenital diaphragmatic hernia (CDH), is still considered a challenge. The standard application of prosthetic material presents major limitations, such as hernia recurrences in a remarkable number of CDH patients. With this work, we developed a tissue engineering approach based on decellularized diaphragmatic muscle and human cells for the in vitro generation of diaphragmatic-like tissues as a proof-of-concept of a new option for the surgical treatment of large diaphragm defects. A customized bioreactor for diaphragmatic muscle was designed to control mechanical stimulation and promote radial stretching during the construct engineering. In vitro tests demonstrated that both ECM remodeling and fibroblast overgrowth were positively influenced by the bioreactor culture. Mechanically stimulated constructs also increased tissue maturation, with the formation of new oriented and aligned muscle fibers. Moreover, after in vivo orthotopic implantation in a surgical CDH mouse model, mechanically stimulated muscles maintained the presence of human cells within myofibers and hernia recurrence did not occur, suggesting the value of this approach for treating diaphragm defects