The dimorphic transition from the yeast to the filamentous form of growth allows cells to explore their environment for more suitable niches and is often crucial for the virulence of pathogenic fungi. In contrast to their Mep1/3 paralogues, fungal Mep2-type ammonium transport proteins of the conserved Mep-Amt-Rh family have been assigned an additional receptor role required to trigger the filamentation signal in response to ammonium scarcity. Here, genetic, kinetic, expression and structure-function analyses were used to shed light on the poorly characterized signaling role of Saccharomyces cerevisiae Mep2. We show that Mep2 variants lacking the C-terminal tail conserve the ability to induce filamentation, revealing that signaling can proceed in the absence of exclusive binding of putative partners to the largest cytosolic domain of the protein. Our data support that filamentation signaling requires the conformational changes accompanying substrate translocation through the pore crossing the hydrophobic core of Mep2. pHluorin reporter assays show that the transport activity of Mep2 and of non-signaling Mep1 differently affect yeast cytosolic pH in vivo, and that the unique pore variant Mep2H194E, with apparent uncoupling of transport and signaling functions, acquires increased ability of acidification. Functional characterization in Xenopus oocytes reveals that Mep2 mediates electroneutral substrate translocation while Mep1 performs electrogenic transport. Our findings highlight that the Mep2-dependent filamentation induction is connected to its specific transport mechanism, suggesting a role of pH in signal mediation. We also show that the signaling process is conserved for the Mep2 protein from the human pathogen Candida albicans. Our results allow to propose a model for the sensing function of Mep2 where pH and calcium are key players. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

While ammonium is an excellent nitrogen source for microorganisms and plants, it is known as acytotoxic metabolite and for its critical role in acid/base homeostasis in animals. Ammonium transportinside the cells is ensured by proteins of the Mep-Amt-Rh superfamily, which are conserved frombacteria to humans.The main objective of the thesis is to refine the understanding of the regulation of the three ammoniumtransport proteins Mep1, Mep2 and Mep3 from Saccharomyces cerevisiae. The three Mep proteins areregulated by the Npr1 kinase and the conserved TORC1 signaling pathway. While the activity of Mep2is regulated by phosphorylation of the C-terminal 457 serine, the activity of Mep1 and Mep3 is inhibitedby the factor Amu1 / Par32. In the presence of a poor nitrogen source, Npr1 induces phosphorylation ofAmu1 which appears mainly cytosolic and, Mep1 and Mep3 are active. On the other hand, in thepresence of a good nitrogen source, the activity of TORC1 induces the inhibition of Npr1 and thereforethe dephosphorylation of Amu1 which accumulates at the cell surface and inactivates Mep1 and Mep3.In order to further study the regulation of Mep1 / 3, a genetic screen was performed to isolate suppressorsrecovering Mep1-dependent ammonium transport in the absence of Npr1. Several mutations, insertionsand deletions have been identified in the MEP1 and AMU1 genes allowing Mep1 to be activeindependently of Npr1. This work shows that all the point mutations in Mep1 delimit an area at theinterface between the hydrophobic body of Mep1 and the cytosol, and that part of the C-terminus (CTR)is required for optimal activity of Mep1 but appears dispensable for regulation by Amu1 and Npr1. Thegenetic screen also shows that the last 15 amino acids of Amu1 are required to inactivate Mep1. Finally,the isolation of suppressors showing no mutation in MEP1 and AMU1 could reveal new factors involvedin the control of Mep1.The results indicate that Mep1 is inactivated in the presence of glutamine, a good source of nitrogen,and that this inactivation requires Amu1. The glutamine-dependent inactivation of Mep2 was alsostudied in this manuscript. Mass spectrometry analysis revealed putative phosphorylation sites in CTRspecific to the presence or absence of glutamine.This work also addressed the role of Amu1 in the reactivation function of TORC1 after treatment withrapamycin, in particular by confirming that it requires the function of Mep1/3. The study leads to thehypothesis that the transport mechanism specific to Mep1 and Mep3 and different from Mep2 isinvolved in this function.Finally, in order to better understand the mechanisms of regulation and transport of Mep-Amt-Rhproteins, the experimental determination of the three-dimensional structure of different variants ofMep2, in open or closed conformation, and of Mep1 was undertaken. Throughout this work, thecharacterized Mep1 or Mep2 variants were analyzed in silico by using the available three-dimensionalstructures. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

A growing population, urbanization and an increase in the number of industries is causing an increase in sewage sludge (SS) that needs to be either beneficially used or disposed of. Application of SS to agricultural lands is a well-known practice but the plant available phosphate and phosphate fertilizer value of SS has been of concern. This is especially due to the wide variety of soils that SS is applied to and the different phosphate removal processes that sewage waste water undergoes at the water care works and the source used to produce SS. Phosphate phyto-availability and phosphate fertilizer value of petrochemical and municipal wastewater sludges (SS) were determined in four different soils using an incubation study over 168 days, a pot trial over 42 days and a field trial over one season. Phosphate phyto-availability was determined/calculated by means of an incubation approach. Soils were incubated with sludge as well as mono ammonium phosphate (MAP). The soils were then subjected to a Bray-1 extraction after a certain time (42 days, 168 days). The relative phosphate fertilizer value (RPFV) was then expressed as a percentage of the Bray-1 extractability of the sludge-amended soil, relative to the MAP-amended soil. The influence of soil properties, especially clay content, and sludge properties namely phosphate extraction method (chemical and biological) from waste water stream, was investigated to determine the effect on phytoavailability and fertilizer value of phosphate in SS-amended soil. Soil properties were the dominant factor determining plant available phosphate, where plant available phosphate decreased with increasing clay content, irrelevant of the type of treatment. There were, however, significant differences between the chemical and biologically removed SS, where the biologically removed SS had higher plant available phosphate. The RPFV % of the SS was comparable to MAP in terms of its plant availability. Wetting and drying cycles in the pot trial influenced the plant available phosphate from the SS, where the chemically treated sludge showed in general lower plant availability. The RPFV % of the biologically removed phosphate sludges was better than that of MAP and that of the chemically phosphate removed sludge were lower. Application of all the different sludge types resulted in a positive reaction on plant available phosphate for all the soils. All the trials were conducted at pH of about 5.5. However, it is expected that biologically P removed sludge will perform better in acid soils. The reason being that ferric phosphate in the chemically treated sludge is less soluble under pH conditions lower than 5.5 than above it. / Dissertation (MScAgric)--University of Pretoria, 2014. / tm2015 / Plant Production and Soil Science / MScAgric / Unrestricted

<p>Iron oxide is the most common catalyst in solid rocket propellant. We have previously demonstrated increased performance of propellant by encapsulating iron oxide particles within ammonium perchlorate (AP), but only nanoscale particles were used, and encapsulation was only accomplished in fine AP (~20 microns in diameter). In this study, we extended the size of particle inclusions to micron-scale within the AP particles as well the particle sizes of the AP-encapsulated catalyst particles (100s of microns) using fractional crystallization techniques with the AP-encapsulated particles as nucleation sites for precipitation. Here we report catalyst particle inclusions of micron-scale, as well as nanoscale, within AP and present characterization of this encapsulation. Encapsulating micron-sized particles and growing these composite particles could pave the way for numerous possible applications. A study of the thermal degradation of these AP-encapsulated particles compared against a standard mixture of iron oxide and AP showed that AP-encapsulated micron-scale catalyst particles exhibited similar behavior to AP-encapsulated nanoscale particles. Using computed tomography, we found that catalyst particles were dispersed throughout the interior of coarse AP-encapsulated micron-scale catalyst particles and decomposition was induced within these particles around catalyst-rich regions.</p>

The Newmarket Till is a regional aquitard in southern Ontario that overlies the Illinoian to Middle Wisconsinan Lower Sediments and is overlain by the Oak Ridges Moraine (ORM). Geological investigations have mapped the distribution of the till and it is understood that erosional channels, subsequently infilled with fluvial material, breach the till and may create enhanced hydraulic connection between overlying and underlying aquifers. However, little is known about the protective capability of the Newmarket Till where it is intact. This study used natural tracers to assess the extent of transport in the aquitard-aquifer system. Stable isotopes of water (δ18O and δ2H) showed a depletion trend versus depth. In the Newmarket Till most of the samples had isotope ratios similar to meteoric water data from the nearest location (Egbert, ON). The depleted values of δ18O in the Thorncliffe Formation suggest a remnant signature of early-Holocene precipitation (-16‰ at the depth of 60 m). Elevated levels of NO3- and Cl- were detected near the surface and because of the low permeability aquitard (Newmarket Till), they could not migrate to depth. Total extractable ammonium concentrations are ranging from 4.09 ppm from near the surface to 60 ppm in the lowest part of the Newmarket and then gradually increase to 514 ppm in the bottom of the Thorncliffe Formation. The combination of high NH4+ values and organic carbon content in the Thorncliffe Formation suggests a natural source from mineralization of organic N. The fractionation which happened between δ15Nsediment and δ15N-NH4 may have three explanations: (1) lighter isotopes diffuse faster than heavier ones, so the higher rate of diffusion can cause fractionation; (2) heavier isotopes partition to exchange sites, causing fractionation along the transport pathway; (3) dissociation of NH4+ to NH3 under anaerobic condition. Positive values for δ13C in groundwater in the Thorncliffe Formation are likely due to i) a contribution of carbonate mineral dissolution, and ii) methanogenesis - the Archea favor the lighter isotope of C (12C). Methanogenesis, therefore, enriches the δ13C-DIC was enriched; however, the δ13C in dissolved organic carbon (DOC) is depleted. These geochemical characteristics demonstrate a long residence time for the porewater in the system and indicate that the Newmarket till inhibits recharge of recent precipitation, thereby providing protection to the underlying aquifers from surface-derived contaminants.

Resumo: O nitrogênio é considerado alimento de massa, isto é, o elemento químico que as plantas geralmente necessitam em maior quantidade principalmente na fase ativa de crescimento. Entretanto, este nutriente normalmente, confere às plantas maior ciclo vegetativo. A adubação nitrogenada em condições normais de cultivo em conjunto com a produção endógena de giberelinas, acarreta maior alongamento de hastes florais, que muitas vezes ganham dimensões em altura, não acompanhada de aumento no diâmetro ou número de flores. No caso do gladíolo, cujo objetivo comercial é a produção de hastes de maior tamanho, entretanto, com qualidade floral, o ethiltrinezapac poderia ser uma opção, uma vez que atua com o inibidor da síntese de giberelina, sendo assim, teoricamente, o nitrogênio ao invés de atuar exclusivamente no crescimento em extensão associado a este regulador, aumentaria além do comprimento, também o diâmetro das hastes florais e com consequente aumento do número e tamanho das flores. Desta forma, o presente trabalho teve como objetivo avaliar a produção e qualidade de gladíolo, em função de modos de aplicação de nitrogênio e do regulador de crescimento ethiltrinezapac. Este foi desenvolvido em área irrigada da FEPE - Setor de Produção Vegetal (FE/UNESP), localizada no município de Ilha Solteira (SP) em um ARGISSOLO VERMELHO Distrófico. O gladíolo (variedade white friendship) foi plantado em julho de 2007, em canteiros com 3m de largura por 2,40m de comprimento, com espaçamento de 10 x 50 cm. O delineamento experimental foi de blocos casualizados, em arranjo fatorial 4x2 (4 modos de aplicação de nitrogênio (a) 90 kg ha-1 de N (estádio de 2-3 folhas das plantas de gladíolo); b) 90 kg ha-1 de N (estádio de emissão das inflorescências); c) 45 kg ha-1 de N (estádio de 2-3 folhas das plantas de gladíolo) + 45 kg ha-1 de N (estádio de emissão... (Resumo completo, click acesso eletrônico abaixo) / Abstract: The nitrogen is considered food mass, that is, the element chemical plants generally require more active phase primarily in growth. This nutrient, usually, confers to the plants a higger vegetative cycle. The N fertilization in normal crop conditions, together endogenous production of giberelins, causes greater allonge of floral rods, which many times gain dimensions in height and not following increase in the diameter or number of flowers. In the case of Gladiolus, that commercial objective is the production of higher connecting rods, however, with floral quality, ethiltrinezapac could be an option, due to its acting with inhibitor of the giberelin synthesis, being thus, theoretically, nitrogen instead of acting exclusively in the length growth, associate to this regulator would increase beyond the length also the diameter of the floral connecting rods and with consequent increasing the number and size of the flowers. This way, the present study had as objective to evaluate the production and quality of gladiolus, as a function of N and ethiltrinezapac application. This was developed in irrigated area at FEPE - agriculture area belongs to FE/UNESP, located in Ilha Solteira (SP - Brazil) county in a Distrofic Red Argissol. Gladiolus (variety White Friendship) was planted in July of 2007, in seedbeds with 3m width for 2.40m length, with arrangement of 10 x 50 cm. The experimental design was a randomized blocks, in a factorial arrangement 4 x 2 (4 ways of N(a) 90 kg ha-1 of N (stage 2-3 Gladiolus plants sheets); b) 90 kg ha-1 of N (stage of issuance of the inflorescences); c) 45 kg ha-1 of N (stage 2-3 Gladiolus plants sheets) + 45 kg ha-1 of N (stage of issuance of the inflorescences); d) 30 kg ha-1 of N (stage 2-3 Gladiolus plants sheets) + 30 kg ha-1 of N (stage of issuance of the inflorescences) + 30 kg ha-1 of N (15 days after the issue of inflorescence), with and/or without... (Complete abstract click electronic access below) / Orientador: Marcelo Andreotti / Coorientador: Regina Maria Monteiro de Castilho / Banca: Salatiér Buzetti / Banca: Kathia Fernandes Lopes Pivetta / Mestre

Anaerobic ammonium oxidation (anammox) by anammox bacteria contributes significantly to the global nitrogen cycle and plays a major role in sustainable wastewater treatment. To date, autotrophic nitrogen removal by anammox bacteria is the most efficient and environmentally friendly process for the treatment of ammonium in wastewaters; its application can save up to 60% of the energy input, nearly 100% elimination of carbon demand and 80% decrease in excess sludge compared to conventional nitrification/denitrification process. In the anammox process, ammonium (NH4+) is directly oxidized to dinitrogen gas (N2) using intracellular electron acceptors such as nitrite (NO2–) or nitric oxide (NO). In the absence of NO2– or NO, anammox bacteria can couple formate oxidation to the reduction of metal oxides such as Fe(III) or Mn(IV). Their genomes contain homologs of Geobacter and Shewanella cytochromes involved in extracellular electron transfer (EET). However, it is still unknown whether anammox bacteria have EET capability and can couple the oxidation of NH4+ with transfer of electrons to extracellular electron acceptors. In this dissertation, I discovered by using complementary approaches that in the absence of NO2–, freshwater and marine anammox bacteria couple the oxidation of NH4+ with transfer of electrons to carbon-based insoluble extracellular electron acceptors such as graphene oxide (GO) or electrodes poised at a certain potential in microbial electrolysis cells (MECs). Metagenomics, fluorescence in-situ hybridization and electrochemical analyses coupled with MEC performance confirmed that anammox electrode biofilms were responsible for current generation through EET-dependent oxidation of NH4+. 15N-labelling experiments revealed the molecular mechanism of the EET-dependent anammox process. NH4+ was oxidized to N2 via hydroxylamine (NH2OH) as intermediate when electrode was used as the terminal electron acceptor. Comparative transcriptomics analysis supported isotope labelling experiments and revealed an alternative pathway for NH4+ oxidation coupled to EET when electrode was used as electron acceptor. The results presented in my dissertation provide the first experimental evidence that marine and freshwater anammox bacteria can couple NH4+ oxidation with EET, which is a significant breakthrough that is promising in the context of implementing EET-dependent anammox process for energy-efficient treatment of nitrogen using bioelectrochemical systems.

The principal aim of this research was to determine the optimum conditions of extraction and separation of niobium and tantalum with octanol as solvent, from Mozambican tantalite using ammonium bifluoride as an alternative to hydrofluoric acid. The extraction of niobium and tantalum from tantalite can be divided into three activities, viz., acid treatment of the ore to bring the niobium and tantalum values into solution, separation of niobium and tantalum by solvent extraction and preparation of pure niobium pentoxide and tantalum pentoxide by precipitation followed by calcination. An initial solution was prepared by melting a mixture of tantalite and ammonium bifluoride followed by leaching of the soluble component with water and separation of the solution by filtration. The solution filtered was successfully used after adjustment of the acidity for the extraction and separation of niobium and tantalum. After liquid-liquid extraction highly pure niobium pentoxide and tantalum pentoxide were obtained through precipitation with ammonium hydroxide and calcination. Comparative experiments were performed modifying the following variables: decomposition temperature; decomposition time of the digestion of niobium and tantalum; acid concentration of feed solution; solvent agent; and stripping agent. From the experimental results it was determined that the decomposition temperature, decomposition time of the digestion of niobium and tantalum, the acidity of the feed solution, the solvent agent, and stripping agent, all have an important effect on the extraction and separation of tantalum and niobium. The optimal conditions were determined to be: tantalite-to-ammonium bifluoride 1:30; decomposition temperature 250°C; decomposition time 3 hours; and a water leach period of 10 minutes. Under these conditions the leach recovery of niobium and tantalum was about 95.07% and 98.52%, respectively. For nearly complete extraction of tantalum and niobium with 2-octanol, two and three equilibrium stages, respectively, are required. The equilibrium data were obtained at an aqueous-to-organic ratio (A/O) of 1:1 using: 100% 2 octanol; 6 M H2SO4; 10 minutes contact time at room temperature for tantalum; and 100% 2 octanol, 9 M H2SO4, 10 minutes contact time at room temperature for niobium. Different stripping agents were used and water seems to give good result for both. For the nearly complete stripping process four equilibrium stages are required. From the results obtained an optimum stripping efficiency is achieved after 15 minutes for a 1:1 ratio. Niobium and tantalum were then neutralized using 28% ammonium hydroxide. The precipitate obtained was dried and placed in a muffle furnace for 4 hours at 900°C. After calcinations, pure tantalum pentoxide and niobium pentoxide were obtained and characterized using XRD and SEM. Tantalum pentoxide and niobium pentoxide synthesized contain trace impurities such as iron and titanium which can be removed by incorporating an appropriate intermediate treatment. / Dissertation (MSc)--University of Pretoria, 2012. / Chemical Engineering / unrestricted

Cellulose nanocomposites offer interesting potential in terms of improved properties and new functionalities compared with microcomposites. Preparation from colloidal suspensions is promising, since high reinforcement content is possible and a wide range of constituents can be used. In the first study, the challenge is to form a stable suspension of well-dispersed carbon nanotubes (CNT) and nanofibrillated cellulose (NFC) in water and to prepare commingled high CNT content nanopaper structures by filtration. Various surfactants were used to modify CNT. The NFC was stabilized by charged carboxylate groups. A nonylphenol phosphate ester surfactant, NPPE, worked well for CNT and provided a stable and well-dispersed water suspension of CNT and NFC. Field emission scanning electron microscopy (FE-SEM), porosimetry and atomic force microscopy (AFM) were used to characterize nanopaper structure, and tensile properties were measured as well as surface resistivity. The processing route is water based and it is possible to prepare thin coatings as well as thicker films with a combination of low surface resistivity, flexibility in bending and high strength and toughness in tension.  As inspired by organo-modified layered silicates, the objective of the second study is to develop an environmentally friendly procedure for the surface modification of cellulose nanocrystals, CNC, using quaternary ammonium salts via adsorption. In order to obtain higher surface charge density on CNC, a new route is developed for preparation of CNC with carboxylic acid groups. Quanternary ammonium cations bearing alkyl, phenyl, glycidyl, and diallyl groups are used to modify CNC to render their surface more hydrophobic. The structure and surface hydrophobicity of unmodified and modified CNC as well as their dispersibility in organic solvent are characterized by AFM, FE-SEM, Fourier-transformed infrared spectroscopy (FT-IR), X-Ray analysis (XDR) and contact angle measurement (CAM). Future work will focus on surface-modified nanocelluloses in composite materials, in order to learn more about surface treatment effects on nanocomposite properties. / Nanokompositer från cellulosa har potential att ge starkt förbättrade egenskaper och ny funktionalitet jämfört med mikrokompositer. De ger även möjlighet till komposittillverkning från kolloidala suspensioner där man kan uppnå hög halt av förstärkningsfasen. Det är också möjligt att välja från en bred flora av lösliga och dispergerbara materialkomponenter. I första studien är utmaningen att skapa en stabil och väldispergerad suspension av kolnanorör (CNT) och nanofibrillerad cellulosa (NFC) i vatten för att genom filtrering framställa nanopapper med interpenetrerande CNT och NFC nätverk. Olika ytaktiva ämnen användes för att modifiera CNT. NFC stabiliserades genom laddade karboxylgrupper på ytan. En nonylfenol fosfatester, NPPE, fungerade bra för CNT och resulterade i en stabil och väldispergerad vattensuspension av CNT och NFC. FE-SEM, densitometri och AFM användes för att karakterisera nanopapperstruktur. Mekaniska egenskaper och ytresistivitet mättes. Processen för framställning av CNT/NFC nanopapper är vattenbaserade och det är möjligt att framställa tunna ytbeläggningar likväl som tjockare filmer. Dessa strukturer har en kombination av låg resistivitet, flexibilitet i böjning liksom hög hållfasthet och seghet i dragbelastning.  Syftet med den andra studien är att utgå från organo-modifierade skiktade silikater (leror) för att utveckla en miljövänlig ytmodifieringsmetod för nanocellulosa. För att öka ytladdningstätheten på CNC (nanokristaller från cellulosa) utvecklas ett nytt sätt att skapa karboxylgrupper på ytan. Kvarternära ammoniumsalter med alkyl, fenyl, glycidyl och diallylgrupper används för att göra ytan på CNC mer hydrofob. Ytans struktur och hydrofoba karaktär, liksom dispersionsegenskaper i organiska lösningsmedel, karakteriseras med hjälp av AFM, FE-SEM, FT-IR, XDR och kontaktvinkelmätning. Fortsatt arbete kommer att fokusera på ytmodifierad cellulosa i kompositmaterial, för att utveckla förståelsen för effekter av ytmodifiering på nanokompositers egenskaper / <p>QC 20120302</p>

Anammox is one of the main processes discovered quite recently for removal of ammonium from wastewater. Anammox process is cost effective, in that low energy and carbon source is needed. Partial nitritation is a perquisite for anammox in wastewater treatment for removal nitrogen and therefore partial nitritation/Anammox technology is studied substantially and applied in full-scale. However, the technology at present can only be used to treat high rich ammonium streams. Application of Anammox for treatment of low ammonium wastewater is not possible because of low yield of Anammox bacteria. The study aimed at devising strategies for using the Anammox technology to treat wastewater streams with low concentration of ammonium nitrogen. The objective was to get systems that could concentrate ammonium from low ammonium waste streams, so as to be able to treat it with partial nitritation/Anammox process. Two methods were used to concentrate ammonium: ion exchange and reverse osmosis. Ion exchange method was used to concentrate UASB effluents of about 24 - 40 mg NH4-N/l to 188 - 367 mg  NH4-N/l respectively which is about 9 times the initial concentrations. At VRF 5, 163 mg  NH4-N/l concentrate was attained from 41.8 mg  NH4-N/l RO feed. Results also showed that concentrates from both methods are able to be treated with partial nitritation/Anammox technology. However it took more than 32 hours to complete treatment of ion exchange concentrates while it took less than 24 hours to finish the partial nitritation/Anammox process of RO concentrates. The longer time taken can be attributed to high salinity of the concentrates which is as a result of NaCl which was used for regeneration in ion exchange process. Both ion exchange and reverse osmosis are viable methods for concentrating ammonium from UASB effluents. Dissolved oxygen was very important factor that influenced the biological process.