The adjuvant activity of two urea derivatives on cytokinins: an example of serendipitous dual effect

The aim of this study was to investigate the action spectrum of two urea derivatives, the 1,3-di(benzo[d]oxazol-5-yl)urea (5-BDPU) and the 1,3-di(benzo[d]oxazol-6-yl)urea (6-BDPU). In order to evaluate a possible adjuvant activity on cytokinins the compounds alone or in the simultaneous presence of different cytokinins were assayed either on in vitro typical cytokinin-related bioassays, or on in planta interaction with cytokinin signal transduction pathway. The compounds ability to activate the cytokinin receptor CRE1/AHK4 was studied either by a heterologous bacterial assay or by a competitive binding assay and docking simulations were performed with the crystal structure of the same receptor. Then, owing to their chemical structure which resembles that of urea-type cytokinins, the ability of 5- and 6-BDPU to inhibit the activity of cytokinin oxidase/dehydrogenase of Zea mays (ZmCKX1) was investigated and docking simulations were performed as well. Accordingly to the experimental results, we speculate that BDPUs could show a dual activity: the blocking of the conformational re-adaption of CRE1/AHK4 receptor maintaining the cytokinin inside its binding pocket, thus possibly enhancing its kinase action; the inhibition of cytokinin oxidase/dehydrogenase activity thus possibly preventing its cleavage of natural cytokinins with isoprenoid side chain. Graphic abstract: [Figure not available: see fulltext.

Irradiation programme HFR phase IIb - SPICE. Impact testing on up to 16.3 dpa irradiated RAFM steels. Final report for task TW2-TTMS 001b-D05

Bestrahlungsprogramm HFR Phase IIb – SPICE: Kerbschlagbiegeuntersuchungen an bis 16.3 dpa bestrahlten RAFM Stählen Die Untersuchung des bestrahlungsinduzierten Versprödungsverhaltens von niedrigaktivierbaren ferritisch-martensitischen (RAFM) Stählen bildet das Ziel der vorliegenden Arbeit. Die Neutronenbestrahlung bei verschiedenen Bestrahlungstemperaturen (250-450 °C) bis zu einer Schädigung von 16,3 dpa (nominell) wurde im Hoch Fluss Reaktor („High Flux Reactor“) Petten im Rahmen des HFR Phase IIb (SPICE) Bestrahlungsprogramms durchgeführt. Die Kerbschlageigenschaften wurden an miniaturisierten Charpy-V Proben (des KLST Typs) mittels der instrumentierten Kerbschlagbiegeversuche ermittelt. Der Hauptschwerpunkt des Bestrahlungsprogramms liegt in der Untersuchung des Einflusses der Neutronenbestrahlung auf die mechanischen Eigenschaften von einem europäischen RAFM Referenzstahl für die Erste Wand des Demonstrationsreaktors (DEMO), EUROFER97, bei zwei verschiedenen Wärmebehandlungen. Die mechanischen Eigenschaften von EUROFER97 wurden mit denen von internationalen Referenzmaterialien (F82H-mod, OPTIFER-Ia, GA3X und MANET-I), welche im SPICE Projekt untersucht wurden, verglichen. Die Bestrahlungsresistenz von bis 16,3 dpa bei 250-450 °C bestrahltem EUROFER97 ist vergleichbar mit der von besten Referenz-Strukturmaterialien. Es wurde für alle untersuchten RAFM Stähle eine erhebliche Tieftemperaturversprödung (Tirr ≤ 300 °C) beobachtet. Die Wärmebehandlung von EUROFER97 bei höherer Austenisierungstemperatur führt zur wesentlichen Verbesserung des Versprödungsverhaltens bei niedrigen Bestrahlungstemperaturen (Tirr ≤ 350 °C). Bei Tirr ≥ 350 °C liegen die Sprödbruchübergangstemperaturen (DBTT) von untersuchten niedrigaktivierbaren Stählen unterhalb von -20 °C und damit weit unterhalb der Anwendungstemperatur. Die Analyse des Verfestigungs- vs. Versprödungs- Verhaltens deutet auf die verfestigungsdominierte Versprödung unterhalb von Tirr ≤ 350 °C mit 0,17≤ C100 ≤ 0,53 °C/MPa. Oxiddispersionsgehärteter ODS EUROFER Stahl mit 0,5 wt.% Y2O3 wurde bei ausgewählten Temperaturen bestrahlt. Bereits im unbestrahlten Zustand zeigte ODS EUROFER keine be- friedigende Kerbschlagbiegeeigenschaften, gekennzeichnet durch eine niedrige USE = 2,54 J und durch eine große DBTT = 135 °C. Die Zunahme von USE für Bestrahlungstemperaturen unterhalb von Tirr ≤ 350°C deutet außerdem auf ein nicht optimiertes Herstellungsverfahren. Bei Tirr = 250 °C ist die bestrahlungsinduzierte Verschiebung der Übergangstemperatur vergleichbar mit der vom EUROFER97. Bei höheren Bestrahlungstemperaturen zeigt hingegen ODS EUROFER größere Versprödung im Vergleich zum Referenzmetall. Die Rolle des Heliums in der Materialversprödung wurde an EUROFER97 basierten Stählen untersucht, welche mit verschiedenem Bor Gehalt (0,008-0,112 wt.%) dotiert wurden. Bor dotierte Proben zeigen zunehmende Versprödung und Abnahme der Zähigkeit mit der Zunahme des generierten Helium Gehalts. Bis zu einem Helium Gehalt von 84 appm ist bei Tirr = 250 °C Helium induzierte Versprödung auf Helium induzierte Verfestigung zurückzuführen. Höhere Helium Konzentrationen führten zu weiteren Versprödungsmechanismen zusätzlich zur Helium induzierten Verfestigung

Immobilization of the Glycosylphosphatidylinositol-anchored Gas1 protein into the chitin ring and septum is required for proper morphogenesis in yeast

Gas1p is a glucan-elongase that plays a crucial role in yeast morphogenesis. It is predominantly anchored to the plasma membrane through a glycosylphosphatidylinositol but a fraction was also found covalently bound to the cell wall. We have used fusions with the green or red fluorescent proteins (GFP or RFP) to determine its localization. Gas1p was present in microdomains of the plasma membrane, at the mother-bud neck and in the bud scars. By exploiting the instability of RFP-Gas1p we identified mobile and immobile pools of Gas1p. Moreover, in chs3\uf044 cells the chitin ring and the cross-linked Gas1p were missing but this unveiled an additional unexpected localization of Gas1p along the septum line in cells at cytokinesis. Localization of Gas1p was also perturbed in a chs2\uf044 mutant where a remedial septum is produced. Phenotypic analysis of cells expressing a fusion of Gas1p to a trans-membrane domain unmasked new roles of the cell wall-bound Gas1p in the maintenance of the bud neck size and in cell separation. We present evidence that the Crh1p and Crh2p are required for tethering Gas1p to the chitin ring and bud scar. These results reveal a new mechanism of protein immobilization at specific sites of the cell envelop

Different pioneer plant species select specific rhizosphere bacterial communities in a high mountain environment

The rhizobacterial communities of 29 pioneer plants belonging to 12 species were investigated in an alpine ecosystem to assess if plants from different species could select for specific rhizobacterial communities. Rhizospheres and unvegetated soils were collected from a floristic pioneer stage plot at 2,400 m a.s.l. in the forefield of Weisskugel Glacier (Matsch Valley, South Tyrol, Italy), after 160 years of glacier retreat. To allow for a culture-independent perspective, total environmental DNA was extracted from both rhizosphere and bare soil samples and analyzed by Automated Ribosomal Intergenic Spacer Analysis (ARISA) and Denaturing Gradient Gel Electrophoresis (DGGE). ARISA fingerprinting showed that rhizobacterial genetic structure was extremely different from bare soil bacterial communities while rhizobacterial communities clustered strictly together according to the plant species. Sequencing of DGGE bands showed that rhizobacterial communities were mainly composed of Acidobacteria and Proteobacteria whereas bare soil was colonized by Acidobacteria and Clostridia. UniFrac significance calculated on DGGE results confirmed the rhizosphere effect exerted by the 12 species and showed different bacterial communities (P < 0.05) associated with all the plant species. These results pointed out that specific rhizobacterial communities were selected by pioneer plants of different species in a high mountain ecosystem characterized by oligotrophic and harsh environmental conditions, during an early primary succession

Safe-site effects on rhizosphere bacterial communities in a high-altitude alpine environment

The rhizosphere effect on bacterial communities associated with three floristic communities (RW, FI, and M sites) which differed for the developmental stages was studied in a high-altitude alpine ecosystem. RW site was an early developmental stage, FI was an intermediate stage, M was a later more matured stage. The N and C contents in the soils confirmed a different developmental stage with a kind of gradient from the unvegetated bare soil (BS) site through RW, FI up to M site. The floristic communities were composed of 21 pioneer plants belonging to 14 species. Automated ribosomal intergenic spacer analysis showed different bacterial genetic structures per each floristic consortium which differed also from the BS site. When plants of the same species occurred within the same site, almost all their bacterial communities clustered together exhibiting a plant species effect. Unifrac significance value (P < 0.05) on 16S rRNA gene diversity revealed significant differences (P < 0.05) between BS site and the vegetated sites with a weak similarity to the RW site. The intermediate plant colonization stage FI did not differ significantly from the RW and the M vegetated sites. These results pointed out the effect of different floristic communities rhizospheres on their soil bacterial communities

Proceedings of the Workshop on Monte Carlo's, Physics and Simulations at the LHC PART II

These proceedings collect the presentations given at the first three meetings of the INFN "Workshop on Monte Carlo's, Physics and Simulations at the LHC", held at the Frascati National Laboratories in 2006. The first part of these proceedings contains pedagogical introductions to several basic topics of both theoretical and experimental high pT LHC physics. The second part collects more specialised presentations.Comment: 157 pages, 136 figures; contribution by M. Grazzini has been adde

Mineral\u2013microbe interactions : Biotechnological potential of bioweathering

Mineral\u2013microbe interaction has been a key factor shaping the lithosphere of our planet since the Precambrian. Detailed investigation has been mainly focused on the role of bioweathering in biomining processes, leading to the selection of highly efficient microbial inoculants for the recovery of metals. Here we expand this scenario, presenting additional applications of bacteria and fungi in mineral dissolution, a process with novel biotechnological potential that has been poorly investigated. The ability of microorganisms to trigger soil formation and to sustain plant establishment and growth are suggested as invaluable tools to counteract the expansion of arid lands and to increase crop productivity. Furthermore, interesting exploitations of mineral weathering microbes are represented by biorestoration and bioremediation technologies, innovative and competitive solutions characterized by economical and environmental advantages. Overall, in the future the study and application of the metabolic properties of microbial communities capable of weathering can represent a driving force in the expanding sector of environmental biotechnology

Rhizosheath-root system changes exopolysaccharide content but stabilizes bacterial community across contrasting seasons in a desert environment

Abstract Background In hot deserts daily/seasonal fluctuations pose great challenges to the resident organisms. However, these extreme ecosystems host unique microenvironments, such as the rhizosheath–root system of desert speargrasses in which biological activities and interactions are facilitated by milder conditions and reduced fluctuations. Here, we examined the bacterial microbiota associated with this structure and its surrounding sand in the desert speargrass Stipagrostis pungens under the contrasting environmental conditions of summer and winter in the Sahara Desert. Results The belowground rhizosheath–root system has higher nutrient and humidity contents, and cooler temperatures than the surrounding sand. The plant responds to the harsh environmental conditions of the summer by increasing the abundance and diversity of extracellular polymeric substances (EPS) compared to the winter. On the contrary, the bacterial community associated with the rhizosheath–root system and its interactome remain stable and, unlike the bulk sand, are unaffected by the seasonal environmental variations. The rhizosheath–root system bacterial communities are consistently dominated by Actinobacteria and Alphaproteobacteria and form distinct bacteria communities from those of bulk sand in the two seasons. The microbiome-stabilization mediated by the plant host acts to consistently retain beneficial bacteria with multiple plant growth promoting functions, including those capable to produce EPS, which increase the sand water holding capacity ameliorating the rhizosheath micro-environment. Conclusions Our results reveal the capability of plants in desert ecosystems to stabilize their below ground microbial community under seasonal contrasting environmental conditions, minimizing the heterogeneity of the surrounding bulk sand and contributing to the overall holobiont resilience under poly-extreme conditions

Rhomboid family member 2 regulates cytoskeletal stress-associated Keratin 16.

Keratin 16 (K16) is a cytoskeletal scaffolding protein highly expressed at pressure-bearing sites of the mammalian footpad. It can be induced in hyperproliferative states such as wound healing, inflammation and cancer. Here we show that the inactive rhomboid protease RHBDF2 (iRHOM2) regulates thickening of the footpad epidermis through its interaction with K16. K16 expression is absent in the thinned footpads of irhom2-/- mice compared with irhom2+/+mice, due to reduced keratinocyte proliferation. Gain-of-function mutations in iRHOM2 underlie Tylosis with oesophageal cancer (TOC), characterized by palmoplantar thickening, upregulate K16 with robust downregulation of its type II keratin binding partner, K6. By orchestrating the remodelling and turnover of K16, and uncoupling it from K6, iRHOM2 regulates the epithelial response to physical stress. These findings contribute to our understanding of the molecular mechanisms underlying hyperproliferation of the palmoplantar epidermis in both physiological and disease states, and how this 'stress' keratin is regulated