Structures of substrate- and nucleotide-bound propionate kinase from Salmonella typhimurium: substrate specificity and phosphate-transfer mechanism

Kinases are ubiquitous enzymes that are pivotal to many biochemical processes. There are contrasting views on the phosphoryl-transfer mechanism in propionate kinase, an enzyme that reversibly transfers a phosphoryl group from propionyl phosphate to ADP in the final step of non-oxidative catabolism of L-threonine to propionate. Here, X-ray crystal structures of propionate- and nucleotide-bound Salmonella typhimurium propionate kinase are reported at 1.8-2.0 angstrom resolution. Although the mode of nucleotide binding is comparable to those of other members of the ASKHA superfamily, propionate is bound at a distinct site deeper in the hydrophobic pocket defining the active site. The propionate carboxyl is at a distance of approximate to 5 angstrom from the -phosphate of the nucleotide, supporting a direct in-line transfer mechanism. The phosphoryl-transfer reaction is likely to occur via an associative S(N)2-like transition state that involves a pentagonal bipyramidal structure with the axial positions occupied by the nucleophile of the substrate and the O atom between the - and the -phosphates, respectively. The proximity of the strictly conserved His175 and Arg236 to the carboxyl group of the propionate and the -phosphate of ATP suggests their involvement in catalysis. Moreover, ligand binding does not induce global domain movement as reported in some other members of the ASKHA superfamily. Instead, residues Arg86, Asp143 and Pro116-Leu117-His118 that define the active-site pocket move towards the substrate and expel water molecules from the active site. The role of Ala88, previously proposed to be the residue determining substrate specificity, was examined by determining the crystal structures of the propionate-bound Ala88 mutants A88V and A88G. Kinetic analysis and structural data are consistent with a significant role of Ala88 in substrate-specificity determination. The active-site pocket-defining residues Arg86, Asp143 and the Pro116-Leu117-His118 segment are also likely to contribute to substrate specificity.

Strength reliability and in vitro degradation of three-dimensional powder printed strontium-substituted magnesium phosphate scaffolds

Strontium ions (Sr2+) are known to prevent osteoporosis and also encourage bone formation. Such twin requirements have motivated researchers to develop Sr-substituted biomaterials for orthopaedic applications. The present study demonstrates a new concept of developing Sr-substituted Mg-3(PO4)(2) - based biodegradable scaffolds. In particular, this work reports the fabrication, mechanical properties with an emphasis on strength reliability as well as in vitro degradation of highly biodegradable strontium-incorporated magnesium phosphate cements. These implantable scaffolds were fabricated using three-dimensional powder printing, followed by high temperature sintering and/or chemical conversion, a technique adaptable to develop patient-specific implants. A moderate combination of strength properties of 36.7 MPa (compression), 242 MPa (bending) and 10.7 MPa (tension) were measured. A reasonably modest Weibull modulus of up to 8.8 was recorded after uniaxial compression or diametral tensile tests on 3D printed scaffolds. A comparison among scaffolds with varying compositions or among sintered or chemically hardened scaffolds reveals that the strength reliability is not compromised in Sr-substituted scaffolds compared to baseline Mg-3(PO4)(2). The micro-computed tomography analysis reveals the presence of highly interconnected porous architecture in three-dimension with lognormal pore size distribution having median in the range of 17.74-26.29 mu m for the investigated scaffolds. The results of extensive in vitro ion release study revealed passive degradation with a reduced Mg2+ release and slow but sustained release of Sr2+ from strontium-substituted magnesium phosphate scaffolds. Taken together, the present study unequivocally illustrates that the newly designed Sr-substituted magnesium phosphate scaffolds with good strength reliability could be used for biomedical applications requiring consistent Sr2+-release, while the scaffold degrades in physiological medium. Statement of significance The study investigates the additive manufacturing of scaffolds based on different strontium-substituted magnesium phosphate bone cements by means of three-dimensional powder printing technique (3DPP). Magnesium phosphates were chosen due to their higher biodegradability compared to calcium phosphates, which is due to both a higher solubility as well as the absence of phase changes (to low soluble hydroxyapatite) in vivo. Since strontium ions are known to promote bone formation by stimulating osteoblast growth, we aimed to establish such a highly degradable magnesium phosphate ceramic with an enhanced bioactivity for new bone ingrowth. After post-processing, mechanical strengths of up to 36.7 MPa (compression), 24.2 MPa (bending) and 10.7 MPa (tension) could be achieved. Simultaneously, the failure reliability of those bioceramic implant materials, measured by Weibull modulus calculations, were in the range of 4.3-8.8. Passive dissolution studies in vitro proved an ion release of Mg2+ and PO43- as well as Sr2+, which is fundamental for in vivo degradation and a bone growth promoting effect. In our opinion, this work broadens the range of bioceramic bone replacement materials suitable for additive manufacturing processing. The high biodegradability of MPC ceramics together with the anticipated promoting effect on osseointegration opens up the way for a patient-specific treatment with the prospect of a fast and complete healing of bone fractures. (C) 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

ATP Antagonizes Thrombin-Induced Signal Transduction through 12(S)-HETE and cAMP

15 p.

Origin of Th/U variations in chondritic meteorites

Isotope dilution thorium and uranium analyses of the Harleton chondrite show a larger scatter than previously observed in equilibrated ordinary chondrites (EOC). The linear correlation of Th/U with 1/U in Harleton (and all EOC data) is produced by variation in the chlorapatite to merrillite mixing ratio. Apatite variations control the U concentrations. Phosphorus variations are compensated by inverse variations in U to preserve the Th/U vs. 1/U correlation. Because the Th/U variations reflect phosphate ampling, a weighted Th/U average should converge to an improved solar system Th/U. We obtain Th/U=3.53 (1_-mean=0.10), significantly lower and more precise than previous estimates.

To test whether apatite also produces Th/U variation in CI and CM chondrites, we performed P analyses on the solutions from leaching experiments of Orgueil and Murchison meteorites.

A linear Th/U vs. 1/U correlation in CI can be explained by redistribution of hexavalent U by aqueous fluids into carbonates and sulfates.

Unlike CI and EOC, whole rock Th/U variations in CMs are mostly due to Th variations. A Th/U vs. 1/U linear correlation suggested by previous data for CMs is not real. We distinguish 4 components responsible for the whole rock Th/U variations: (1) P and actinide-depleted matrix containing small amounts of U-rich carbonate/sulfate phases (similar to CIs); (2) CAIs and (3) chondrules are major reservoirs for actinides, (4) an easily leachable phase of high Th/U. likely carbonate produced by CAI alteration. Phosphates play a minor role as actinide and P carrier phases in CM chondrites.

Using our Th/U and minimum galactic ages from halo globular clusters, we calculate relative supernovae production rates for ²³²Th/²³⁸U and ²³⁵U/²³⁸U for different models of r-process nucleosynthesis. For uniform galactic production, the beginning of the r-process nucleosynthesis must be less than 13 Gyr. Exponentially decreasing production is also consistent with a 13 Gyr age, but very slow decay times are required (less than 35 Gyr), approaching the uniform production. The 15 Gyr Galaxy requires either a fast initial production growth (infall time constant less than 0.5 Gyr) followed by very low decrease (decay time constant greater than 100 Gyr), or the fastest possible decrease (≈8 Gyr) preceded by slow in fall (≈7.5 Gyr).

Scientific investigations into the eutrophication of the Norfolk Broads

The word ”Broads” is used to describe a series of relatively shallow lakes resulting from the flooding of medieval peat diggings. Broadland is essentially freshwater, but because the rivers have such low gradients the lower reaches are brackish. The influence of tide is particularly apparent on the River Yare; in Norwich 40 km from the sea there is a vertical movement of half a metre at spring tide. This study examines the problems that the broadlands are facing. The problems are basically the progressive loss of aquatic plants, in particular the macro- phytes, animal life, outbreaks of avian botulism, occasional fish kills due to a toxin produced by the blue-green alga Prymesium parvum and the emergence of very heavy algal blooms. The main factor for the deteriation of the Broaslands is the eutrophication resulting from enhanced nutrient inputs, in particular of nitrates and phosphates, from a variety of sources. The most important of these are sewage effluents, agricultural drainage, which includes fertilisers and nutrient rich effluents from piggeries and dairy un

Al2O3,B2O3对磷酸盐包边玻璃形成区和物化性质影响研究

研究了Al2O3 ,B2O3对P2O5-Al2O3-B2O3-ZnO-Na2O-CuCl玻璃形成区的影响规律；包边玻璃的物化性质随B/(B+Al)的变化规律；以及特定组分包边玻璃的透射光谱和吸收系数。这些研究工作对高功率大尺寸钕磷酸盐激光包边玻璃的研制具有非常重要的意义。

不同氧磷比对磷酸盐包边玻璃性质的影响规律

包边技术是提高大尺寸激光玻璃饱和增益系数的关键技术。采用传统的方法熔制玻璃,研究了 P2O5含量对 P2O5-Al2O3-B2O3-CuCl-Na2O-ZnO磷酸盐包边玻璃的折射率、热膨胀系数、玻璃转变温度、膨胀软化温度以及化学稳定性的影响。结果表明：当 P2O5的摩尔分数为 60%左右,玻璃样品具有最高的折射率（1.522 0）、最低的玻璃转变温度（352.4 ℃）、较好的化学稳定性[0.52 mg/（cm^2·d）]和适宜的热膨胀系数（128.427×10^-7/℃）,是用作钕磷酸盐激光玻璃硬包边的理

Influence of phosphate precursors on the structures of sodium aluminophosphate sols, gels and glasses

Transparent and homogeneous aluminophosphate gels and glasses have been widely synthesized through an aqueous sol-gel route, extending significantly the glass-forming range compared to that accessible via the melt-cooling route. Different phosphorus precursors, sodium polyphosphate (NaPO3) and orthophosphate species (NaH2PO4 and/or H3PO4) were compared with regard to the macroscopic properties and the microscopic structure of the resultant gels and glasses as characterized by extensive high-resolution liquid- and solid-state NMR. Sodium polyphosphate solution results in a substantially wider composition range of homogenous gel formation than orthophosphate solutions, and the two routes produce significant structural differences in the sol and xerogel states. Nevertheless, the structures of the glasses obtained upon gel annealing above 400 degrees C are independent of the P-precursors used. (c) 2007 Elsevier B.V. All rights reserved.

Novel Bi-doped glasses for broadband optical amplification

Broadband infrared luminescence is observed in various Bi-doped oxide glasses prepared by conventional melting-quenching technique. The absorption spectrum of the Bi-doped germanium oxide glass consists of five broad peaks at below 370, 500, 700, 800 and 1000 nm. The fluorescence spectrum exhibits a broad peak at about 1300 nm with full width at half maximum (FWHM) of more than 300 nm when excited by an 808 nm laser diode. The fluorescence lifetime at room temperature decreases with increasing Bi2O3 concentration. Influence of the glass composition and melting atmosphere on the fluorescence lifetime and luminescent intensity is investigated. The mechanism of the broadband infrared luminescence is suggested. The product of stimulated emission cross-section and lifetime of the Bi-doped aluminophosphate glass is about 5.0 X 10(-24) cm(2) s. The glasses might be promising for applications in broadband optical fiber amplifiers and tunable lasers. (c) 2007 Elsevier B.V. All rights reserved.

Variations saisonnières des facteurs hydrologiques et du biovolume microalgal benthique au sein d'un habitat artificiel (acadja-enclos) dans la lagune Ebrié: site d'Adjapoté

This paper presents the results of a comparative study of chemical and physical characteristics (temperature, oxygen, salinity, turbidity, pH, nutrient: phosphates, nitrates, nitrites, ammoniac) within the extensive aquaculture structure (acadja-enclosure or artificial habitats for fishes) and in the surrounding water. Biological characteristics of the periphyton growing on bamboos from acadja-enclosure were also described.

Pig dung as pond manure: effect on water quality, pond productivity and growth of carps in polyculture system

Pig dung was used as manure at 18 and 36 t/ha/year in carp polyculture (without supplementary feeding) for 270 days. It was observed that pig dung at both the levels did not degrade the physico-chemical properties (pH, dissolved oxygen and alkalinity) of water. The nutrient (phosphates and nitrates) level of water was higher in manured ponds than control ponds (no manuring, only supplementary feeding). Further, plankton levels (phyto- and zooplankton) were also significantly higher in manured ponds. The growth of Catla catla and Labeo rohita was significantly more in manured ponds than in control ponds. Growth of Cirrhinus cirrhosus and Cyprinus carpio was significantly more in ponds manured with pig dung at 18 t/ha/year than in control ponds and the growth of Ctenopharyngodon idellus was significantly more in control than in manured ponds.

Aspects of phosphorus pollution from aquaculture

Phosphorus is an essential element for living organisms and exists in waterbodies as dissolved and particulate forms. Phosphorus is required for optimum growth, feed efficiency, bone development and maintenance of acid-base regulation in fish. The presence of high concentration of phosphates in water may indicate presence of pollution as it may accelerate plant growth and disrupt the aquatic ecosystem thereby benefiting certain species and altering species diversity in affected areas. Eutrophication of waterbodies is often correlated with the phosphorus loading into the environment and aquaculture has been identified as one of the sources of phosphorus pollution. Details of the impacts of eutrophication is given in Bernhardt (1981). Phosphorus must be provided in fish feed because of its low concentration in water. Studies made in Europe and Northern America have revealed a phosphorus surplus in most commercial feeds which is above actual requirements; or is supplied in a form which is unavailable to the fish. Surplus phosphorus is excreted, while unavailable phosphorus is passed out in the feces. Discharge of phosphorus from fish farms and hatchery effluents have caused phosphorus pollution in Nordic countries, North America and Europe. This article examines the path of phosphorus pollution, quantification/prediction of phosphorus load from aquaculture and remedial measures.

O uso de fosfatos associado à fitorremediação em solos de Santo Amaro (BA) contaminados por metais tóxicos

A contaminação do solo no município de Santo Amaro (BA) por metais tóxicos provocada pelas atividades da empresa Plumbum Mineração tem gerado impactos sobre a saúde ambiental e humana. O objetivo da presente pesquisa foi avaliar a viabilidade da remediação deste solo contaminado por chumbo e cádmio, utilizando diferentes fontes de fosfatos e fitorremediação com o capim vetiver [Vetiveria zizanioides (L.)]. O estudo foi realizado em colunas de PVC onde amostras de solo foram colocadas com a aplicação de fosfato dihidrogênio de potássio (KH2PO4) (T1); fertilizante fosfato natural reativo (FNR) (T2) e; uma mistura do KH2PO4 e de fertilizante FNR (T3). Amostras de solo contaminado sem tratamento (T0) foram utilizadas como controle. Após 60, 120 e 180 dias, alíquotas do solo foram retiradas das colunas para análises. Ao final de cada período, mudas de capim vetiver [(Vetiveria zizanioides (L.)] foram plantadas em vasos com as amostras de solo: T0, T1, T2 e T3 em triplicata. Para a determinação das concentrações de chumbo e cádmio no solo e tecidos vegetais foi utilizado o ICP-OES. A partir das análises física e química constatou-se que o solo possui textura argilosa e capacidade de troca catiônica (CTC) elevadas. As extrações com solução de ácido dietilenotriaminopentaacético (DTPA) e Toxicity Characteristic Leaching Procedure (TCLP) mostraram que o tratamento T1 seguido do T3 foram os mais eficientes na imobilização de Pb e Cd. Entretanto, todos os tratamentos resultaram em concentrações de metais ainda disponíveis no solo que excediam os limites estabelecidos pela USEPA, sendo o solo, portanto, considerado tóxico mesmo após o tratamento. Com base nas concentrações de metais extraídos através da extração sequencial pelo método BCR após a remediação e a fitorremediação do solo, foi verificado que todos os três tratamentos com fosfatos foram eficientes em imobilizar o Pb e Cd nas formas menos solúveis, porém, o Cd permaneceu mais solúvel e com maior mobilidade do que o Pb. Os ensaios de letalidade utilizando minhoca Eisenia andrei mostraram que a mortalidade observada no solo após 60 dias de tratamento foi significativamente reduzida após 120 e 180 dias de tratamento. A perda de biomassa pelas minhocas também foi reduzida de acordo com o tempo de tratamento. O teste de germinação com alfaces (Lactuca sativa L.) indicou que as amostras de solo tratadas continuam bastante tóxicas, apesar da disponibilidade reduzida do Pb e do Cd como visto nos resultados da extração por TCLP e por BCR. A avaliação de risco ecológico potencial indicou que os tratamentos do solo com fosfatos associado à fitorremediação reduziram a mobilidade do Pb, principalmente nos tratamentos T1 e T3. Para o Cd o risco ecológico potencial aumenta consideravelmente quando comparado com o Pb demonstrando que esse elemento, apesar dos tratamentos com fosfatos mais a fitorremediação continua móvel. O tratamento com KH2PO4 (T1) foi o mais eficiente na redução da mobilidade, disponibilidade e da toxicidade dos metais, seguido pelo T3 e T2 para o Pb e o T3 seguido pelo T1 e T2 para o Cd.

Ocorrência e ecotoxicidade de desreguladores endócrinos químicos em ambientes aquáticos e em sistemas de tratamento de esgoto

A aplicabilidade de um método selecionado de medição indireta de vitelogenina (Vtg) em plasma sanguíneo de peixe, baseado na quantificação de fosfato álcali-lábil (alkali-labile phosphate-ALP) para acessar estrogenicidade em água, foi investigada na presente tese. O método foi originalmente desenvolvido para a espécie de peixe Carassius carassius (Carpa cruciana) e aplicado pela primeira vez na espécie Oreochromis niloticus (Tilápia do Nilo) no presente estudo. Com o objetivo de acessar a sensibilidade do método, em uma primeira etapa da investigação foram realizados estudos laboratoriais com soluções estoques de 17-ethinylestradiol (EE2), 17-estradiol (E2), e estrona (E1). Os efeitos destes hormônios foram investigados com base tanto na concentração quanto na carga, utilizando-se para tanto, unidades experimentais com volumes distintos (2 L e 130 L). Após a validação do método de ALP, a estrogenicidade foi avaliada nas seguintes águas contaminadas: (i) afluente e efluente de uma grande estação de tratamento de esgotos convencional (ETE) e de uma estação descentralizada de tratamento de esgoto de pequeno porte (Ecossistema Engenheirado-DEE); (ii) água superficial (SW) e água subterrânea (GW) coletadas em uma área de brejo contaminada com gasolina; (iii) água de uma lagoa urbana (LRF) da cidade do Rio de Janeiro, com alta densidade populacional e descarte clandestino de esgoto. Na segunda etapa foram analisados em microalgas os efeitos (outros que não disrupção endócrina) causados pelos hormônios EE2, E2 e E1. Os hormônios foram testados individualmente e em misturas, em culturas individuais e combinada (S+) das espécies de microalgas unicelulares P. subcapitata e D. subspicatus. Com base nos níveis de ALP para a espécie de peixe e no EC50 para as espécies de algas, os resultados mostraram que o EE2 e o E2 causaram disrupção endócrina superior e foram mais tóxicos do que o E1 para peixes e microalgas respectivamente. Quando em misturas (E+) de concentrações equivalentes (EE2:E2:E1), os estrogênios resultaram em efeito aditivo para as espécies O. niloticus e P. subcapitata, e menos que aditivo para D. subspicatus e cultivo misto de algas (S+). Culturas contendo ambas as espécies de algas (S+) por um longo período de exposição (96 h) resultaram na atenuação dos efeitos tóxicos causados pela exposição, tanto individual (EE2, E2 ou E1), quanto na mistura (E+) dos estrogênios, medidos em termos de EC50 (T0h 0,07; 0,09; 0,18; e 0,06 g mL-1; e T96h 1,29; 1,87; 5,58; e 4,61 g mL-1, respectivamente). O DEE apresentou uma maior eficiência na remoção dos disrutores endócrinos do que a ETE convencional. Foi detectada estrogenicidade em amostras da LRF, e de água SW e GW em área brejosa contaminada com gasolina. Os resultados dos ensaios sugerem que as interações (efeitos aditivos ou menos que aditivo) causadas pela mistura dos estrogênios assim como, as interações entre as espécies de algas afetaram o resultado final dos ensaios ecotoxicológicos. Um fator raramente abordado em estudos ecotoxicológicos que foi destacado na presente tese refere-se à importância de considerar não somente a concentração e a dosagem, mas também a carga aplicada e o volume das unidades experimentais. Devido à boa sensibilidade do O. niloticus quando exposto às concentrações relativamente baixas dos estrogênios, a combinação do método de ALP com os biomarcadores auxiliares (particularmente MN) pode ser um protocolo adequado para a detecção de estogenicidade e genotoxicidade respectivamente em diferentes ambiente aquáticos contaminados, como parte de um programa de monitoramento ambiental

Studies on water quality and fish diversity of Sadatpur lake in Ahmednagar district Maharashtra

The present study deals with the water quality and ichthyofauna of Sadatpur lake. Monthly changes in water quality parameters such as temperature, pH, dissolved oxygen (DO), total alkalinity, chlorides, sulphates, phosphates and nitrites have been studied for a period of one year beginning from January 2005 and were investigated to assess the suitability of this lake for fish and fisheries practices. All the physico-chemical parameters determined, revealed that the fluctuations in water temperature, pH, dissolved oxygen, alkalinity and nitrites, were within the desirable limits. On the other hand, chloride and sulphate were lower whereas phosphate was higher than the desirable limits. Altogether 24 fish species belonging to 17 genera and 2 orders were found to be present in the lake. Among fish species family Cyprinidae dominated the lake. The lake subsistence fishery served the need of local fishermen community.