950 resultados para Beckman pH meter
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RESUME: Etude de l'activation et de l'inactivation pH-dépendantes des canaux ASICs (Acid-Sensing Ion Channels) Benoîte BARGETON, Département de Pharmacologie et de Toxicologie, Université de Lausanne, rue du Bugnon 27, CH-1005 Lausanne, Suisse Les canaux sodiques ASICs (Acid-Sensing Ion Channels) participent à la signalisation neuronale dans les systèmes nerveux périphérique et central. Ces canaux non voltage dépendants sont impliqués dans l'apprentissage, l'expression de la peur, la neurodégénération consécutive à une attaque cérébrale et la douleur. Les bases moléculaires sous-tendant leur activité ne sont pas encore totalement comprises. Ces canaux sont activés par une acidification du milieu extracellulaire et régulés, entre autres, par des ions tels que le Ca2+, le Zn2+ et le CI". La cristallisation de ASIC inactivé a été publiée. Le canal est un trimére de sous-unités identiques ou homologues. Chaque sous-unité a été décrite en analogie à un avant bras, un poignet et une main constituée d'un pouce, d'un doigt, d'une articulation, une boule β et une paume. Nous avons appliqué une approche bioinformatique systématique pour identifier les pH senseurs putatifs de ASICIa. Le rôle des pH senseurs putatifs a été testé par mutagénèse dirigée et des modifications chimiques combinées à une analyse fonctionnelle afin de comprendre comment les variations de ρ H ouvrent ces canaux. Les pH senseurs sont des acides aspartiques et glutamiques éparpillés sur la boucle extracellulaire suggérant que les changements de pH contrôlent l'activation et l'inactivation de ASIC en (dé)protonant ces résidus en divers endroits de la protéine. Par exemple lors de l'activation, la protonation des résidus à l'interface entre le pouce, la boule β et le doigt d'une même sous-unité induit un mouvement du pouce vers la bouie β et le doigt. De même lors de l'inactivation du canal les paumes des trois sous-unités formant une cavité se rapprochent. D'après notre approche bioinformatique, aucune histidine n'est impliquée dans la détection des variations de pH extracellulaire c'est-à-dire qu'aucune histidine ne serait un pH-senseur. Deux histidines de ASIC2a lient le Zn2+ et modifient l'affinité apparente du canal pour les protons. Une seule des deux est conservée parmi tous les ASICs, hASICIa H163. Elle forme un réseau de liaison hydrogène avec ses voisins conservés. L'étude détaillée de ce domaine, Pinterzone, montre son importance dans l'expression fonctionnelle des canaux. La perturbation de ce réseau par l'introduction d'un résidu hydrophobe (cystéine) par mutagénèse dirigée diminue l'expression du canal à la membrane plasmique. La modification des cystéines introduites par des réactifs spécifiques aux groupements sulfhydryle inhibe les canaux mutés en diminuant leur probabilité d'ouverture. Ces travaux décrivent les effets de l'acidification du milieu extracellulaire sur les canaux ASICs. ABSTRACT: Study of pH-dependent activation and inactivation of ASIC channels Benoîte BARGETON, Department of Pharmacology and Toxicology, University of Lausanne, Rue du Bugnon 27, CH-1G05 Lausanne, Switzerland The ASIC (Acid-Sensing Ion Channels) sodium channels are involved in neuronal signaling in the central and peripheral nervous system. These non-voltage-gated channels are involved in learning, the expression of fear, neurodegeneration after ischemia and pain sensation. The molecular bases underlying their activity are not yet fully understood. ASICs are activated by extracellular acidification and regulated, eg by ions such as Ca2+, the Zn2+ and CI". The crystallization of inactivated ASIC has been published. The channel is a trimer of identical or homologous subunits. Each subunit has been described in analogy to a forearm, wrist and hand consisting of a thumb, a finger, a knuckle, a β-ball and a palm. We applied a systematic computational approach to identify putative pH sensor(s) of ASICIa. The role of putative pH sensors has been tested by site-directed mutagenesis and chemical modification combined with functional analysis in order to understand how changes in pH open these channels. The pH sensors are aspartic and glutamic acids distributed throughout the extracellular loop, suggesting that changes in pH control activation and inactivation of ASIC by protonation / deprotonation of many residues in different parts of the protein. During activation the protonation of various residues at the interface between the finger, the thumb and the β-ball induces the movement of the thumb toward the finger and the β-ball. During inactivation of the channel the palms of the three subunits forming a cavity approach each other. No histidine has been shown to be involved in extracellular pH changes detection, i.e. no histidine is a pH- sensor. Two histidines of ASIC2 bind Zn2+ and alter the apparent affinity of channel for protons. Only one of the two His is conserved among all ASICs, hASICIa H163. This residue is part of a network of hydrogen bonding with its conserved neighbors. The detailed study of this area, the interzone, shows its importance in the functional expression of ASICs. Disturbance of this network by the introduction of hydrophobic residues decreases the cell surface channel expression. Chemical modification of the introduced cysteines by thiol reactive compounds inhibits the mutated channels by a reduction of their open probability. These studies describe the effects of extracellular acidification on ASICs. RESUME GRAND PUBLIC: Etude de l'activation et de l'inactivation pH-dépendantes des canaux ASICs (Acid-Sensing Ion Channels) Benoîte BARGETON, Département de Pharmacologie et de Toxicologie, Université de Lausanne, rue du Bugnon 27, CH-1005 Lausanne, Suisse La transmission synaptique est un processus chimique entre deux neurones impliquant des neurotransmetteurs et leurs récepteurs. Un dysfonctionnement de certains types de synapses est à l'origine de beaucoup de troubles nerveux, tels que certaine forme d'épilepsie et de l'attention. Les récepteurs des neurotransmetteurs sont de très bonnes cibles thérapeutiques dans de nombreuses neuropathologies. Les canaux ASICs sont impliqués dans la neurodégénération consécutive à une attaque cérébrale et les bloquer pourraient permettre aux patients d'avoir moins de séquelles. Les canaux ASICs sont des détecteurs de l'acidité qui apparaît lors de situations pathologiques comme l'ischémie et l'inflammation. Ces canaux sont également impliqués dans des douleurs. Cibler spécifiquement ces canaux permettrait d'avoir de nouveaux outils thérapeutiques car à l'heure actuelle l'inhibiteur de choix, l'amiloride, bloque beaucoup d'autres canaux empêchant son utilisation pour bloquer les ASICs. C'est pourquoi il faut connaître et comprendre les bases moléculaires du fonctionnement de ces récepteurs. Les ASICs formés de trois sous-unités détectent les variations de l'acidité puis s'ouvrent transitoirement pour laisser entrer des ions chargés positivement dans la cellule ce qui active la signalisation neuronale. Afin de comprendre les bases moléculaires de l'activité des ASICs nous avons déterminé les sites de liaison des protons (pH-senseurs), ligands naturels des ASICs et décrit une zone importante pour l'expression fonctionnelle de ces canaux. Grâce à une validation systématique de résultats obtenus en collaboration avec l'Institut Suisse de Bioinformatique, nous avons décrit les pH-senseurs de ASICIa. Ces résultats, combinés à ceux d'autres groupes de recherche, nous ont permis de mieux comprendre comment les ASICs sont ouverts par une acidification du milieu extracellulaire. Une seconde étude souligne le rôle structural crucial d'une région conservée parmi tous les canaux ASICs : y toucher c'est diminuer l'activité de la protéine. Ce domaine permet l'harmonisation des changements dus à l'acidification du milieu extracellulaire au sein d'une même sous-unité c'est-à-dire qu'elle participe à l'induction de l'inactivation due à l'activation du canal Cette étude décrit donc quelle région de la protéine atteindre pour la bloquer efficacement en faisant une cible thérapeutique de choix.
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Alterações no pH da rizosfera de plantas fixadoras de N2 parecem exercer papel fundamental na absorção de micronutrientes que têm sua disponibilidade dependente de alterações da acidez do solo. Estudaram-se variações na absorção de B, Cu, Fe, Mn e Zn durante o ciclo de crescimento e desenvolvimento da soja, induzidas pela fixação biológica de N2 e pelo pH inicial de amostras de dois solos (um LV argiloso e outro arenoso), em um ensaio conduzido em casa de vegetação. Essas amostras foram incubadas com doses de CaCO3 + MgCO3 (4:1) para elevar o pH (H2O) a valores de 5,2, 5,6, 6,2 e 6,6 no solo argiloso e 5,3, 5,6, 5,9 e 6,3 no solo arenoso. Após 60 dias de incubação, essas amostras receberam 450 mg dm-3 de P e 120 mg dm-3 de K no solo. Sementes de soja (Glycine max (L) Merrill), variedade Paranaíba, inoculadas com Bradyrhizobium japonicum, estirpes SEMIA 587 e SEMIA 5019, foram colocadas para germinar. Foram cultivadas quatro plantas por vaso (2,2 dm³) e colhidas aos 16, 20, 24, 28, 32, 36, 40, 46 e 54 dias após a emergência. Determinaram-se o pH da rizosfera (pHr), o pH do solo entre raízes - não rizosférico (pHnr), os teores de B, Cu, Fe, Mn e de Zn na parte aérea e raiz, o N apenas na parte aérea, o número de nódulos e o peso da matéria seca de parte aérea, raiz e nódulos. Observou-se que as mudanças ocorridas no pHr e pHnr foram dependentes do pH inicial dos solos (pHs) e da fixação biológica de N2. O acúmulo de B e de Fe na parte aérea não foi alterado pelos valores de pHr, modificados em função do pHs, exceto para o Fe no solo argiloso. Todavia, aumentos significativos no acúmulo destes nutrientes na parte aérea ocorreram com o aparecimento dos nódulos, a partir de 24 dias após a emergência. Para Cu, Mn e Zn, as diferenças apareceram sobretudo quanto ao pHs. O conteúdo de micronutrientes na planta revelou-se sensível a mudanças no pH rizosférico, principalmente após a nodulação.
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We developed a mathematical model of Ca transport along the late distal convoluted tubule (DCT2) and the connecting tubule (CNT) to investigate the mechanisms that regulate Ca reabsorption in the DCT2-CNT. The model accounts for apical Ca influx across transient receptor potential vanilloid 5 (TRPV5) channels and basolateral Ca efflux via plasma membrane Ca-ATPase pumps and type 1 Na/Ca exchangers (NCX1). Model simulations reproduce experimentally observed variations in Ca uptake as a function of extracellular pH, Na, and Mg concentration. Our results indicate that amiloride enhances Ca reabsorption in the DCT2-CNT predominantly by increasing the driving force across NCX1, thereby stimulating Ca efflux. They also suggest that because aldosterone upregulates both apical and basolateral Na transport pathways, it has a lesser impact on Ca reabsorption than amiloride. Conversely, the model predicts that full NCX1 inhibition and parathyroidectomy each augment the Ca load delivered to the collecting duct severalfold. In addition, our results suggest that regulation of TRPV5 activity by luminal pH has a small impact, per se, on transepithelial Ca fluxes; the reduction in Ca reabsorption induced by metabolic acidosis likely stems from decreases in TRPV5 expression. In contrast, elevations in luminal Ca are predicted to significantly decrease TRPV5 activity via the Ca-sensing receptor. Nevertheless, following the administration of furosemide, the calcium-sensing receptor-mediated increase in Ca reabsorption in the DCT2-CNT is calculated to be insufficient to prevent hypercalciuria. Altogether, our model predicts complex interactions between calcium and sodium reabsorption in the DCT2-CNT.
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Selostus: Korkealla virranvoimakkuudella tainnutettujen broilereiden rintafileen irroitushetken vaikutus lihaksen leikkausvoiman vastukseen, pH:hon, keittohävikkiin ja väriin
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Rice in Rio Grande do Sul State is grown mostly under flooding, which induces a series of chemical, physical and biological changes in the root environment. These changes, combined with the presence of rice plants, affect the availability of exchangeable ammonium (NH4+) and pH of soil solution, whereas the dynamics of both variables can be influenced by soil salinity, a common problem in the coastal region. This study was conducted to evaluate the dynamics of exchangeable NH4+ and pH in the soil solution, and their relation in the solution of Albaqualf soils with different salinity levels, under rice. Four field experiments were conducted with soils with exchangeable Na percentage (ESP) of 5.6, 9.0, 21.2, and 32.7 %. Prior to flooding, soil solution collectors were installed at depths of 5, 10 and 20 cm. The soil solution was collected weekly, from 7 to 91 days after flooding (DAF), to analyze exchangeable NH4+ and pH in the samples. Plant tissue was sampled 77 DAF, to determine N uptake and estimate the contribution of other N forms to rice nutrition. The content of exchangeable NH4+ decreased over time at all sites and depths, with a more pronounced reduction in soils with lower salinity levels, reaching values close to zero. A possible contribution of non-exchangeable NH4+ forms and N from soil organic matter to rice nutrition was observed. Soil pH decreased with time in soils with ESP 5.6 and 9.0 %, being positively correlated with the decreasing NH4+ levels at these sites.
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The use of SMP pH to estimate the potential acidity (H + Al) is more practical than the method of 0.5 mol L-1 calcium acetate for routine laboratory analyses. The objective was to fit an equation to estimate the H + Al from SMP pH values of soils of the State of Pará. From various regions of the state, 177 soil samples were collected, in which the SMP pH was determined in 0.01 mol L-1 CaCl2 solution and H + Al in 0.5 mol L-1 calcium acetate and the results were related by regression analysis. The equation H + Al = 77.77 + 20.61 SMP pH - 1.435 SMP pH² (R² = 0.90) expressed the H + Al values (in cmol c dm-3) best. When the SMP pH values were used in equations referring to other regions or states in Brazil, the H +Al values were over- or underestimated. The potential acidity in soils of Pará can be estimated by the method of SMP pH.
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O desbalanço entre Ca2+, Mg2+ e K+ no solo como consequência das aplicações elevadas de gesso deve-se às relações de tamanho (raio iônico) e densidades de cargas (relação carga/raio) de cada espécie iônica. Quanto maior a densidade de carga, mais intensa será a ligação iônica do cátion com íons de cargas opostas como OH- e SO4-2. Dessa maneira, o uso excessivo de gesso agrícola, sem considerar o balanço de cargas das partículas do solo; o equilíbrio iônico; e a CTC podem resultar em expressiva lixiviação ao longo do perfil do solo. O objetivo deste estudo foi avaliar o efeito de elevadas doses de gesso (0, 7 e 56 t ha-1) nos teores de Ca2+, Mg2+, K+ e pH na solução de um Latossolo Vermelho distrófico cultivado com cafeeiro, obtida pelo método adaptado do extrato aquoso. O solo foi amostrado nas profundidades de 0,15-0,25; 0,35-0,45; 0,75-0,85; 1,15-1,25 e 2,35-2,45 m na linha de plantio, em quatro tratamentos: G-0 - gesso no preparo (aplicação ocorreu em setembro de 2008, distribuído a lanço, na quantidade de 2 t ha-1) e sem gesso na linha de plantio; G-7 - gesso adicionado durante a preparação do solo (2 t ha-1), na mesma condição do G-0 e 7,0 t ha-1 de gesso na linha de plantio; G-56 - gesso adicionado durante a preparação do solo (2 t ha-1), na mesma condição do G-0 e 56 t ha-1 de gesso na linha de plantio (nessas parcelas experimentais as entrelinhas de plantio foram cobertas com braquiária); e CV-7: ausência de braquiária na entrelinha, com gesso no preparo e 7 t ha-1 de gesso na linha, com três repetições distribuídas em blocos ao acaso, totalizando 60 amostras. Após 16 meses da adição de gesso, observou-se redução do pH na solução do solo nas profundidades de 0,15-0,25; 0,35-0,45 e 0,75-0,85 m. A aplicação de gesso agrícola foi eficiente na melhoria do ambiente radicular no subsolo, aumentou a concentração de Mg2+ e Ca2+ na solução do solo, mas reduziu o K+ em profundidade, a partir de 0,85 m. Os teores de Ca2+ e Mg2+ trocáveis na solução do solo estiveram acima do nível crítico; entretanto, os teores de K+ trocável se mantiveram na faixa do valor crítico, indicado para o desenvolvimento da cultura.
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Visible and near infrared (vis-NIR) spectroscopy is widely used to detect soil properties. The objective of this study is to evaluate the combined effect of moisture content (MC) and the modeling algorithm on prediction of soil organic carbon (SOC) and pH. Partial least squares (PLS) and the Artificial neural network (ANN) for modeling of SOC and pH at different MC levels were compared in terms of efficiency in prediction of regression. A total of 270 soil samples were used. Before spectral measurement, dry soil samples were weighed to determine the amount of water to be added by weight to achieve the specified gravimetric MC levels of 5, 10, 15, 20, and 25 %. A fiber-optic vis-NIR spectrophotometer (350-2500 nm) was used to measure spectra of soil samples in the diffuse reflectance mode. Spectra preprocessing and PLS regression were carried using Unscrambler® software. Statistica® software was used for ANN modeling. The best prediction result for SOC was obtained using the ANN (RMSEP = 0.82 % and RPD = 4.23) for soil samples with 25 % MC. The best prediction results for pH were obtained with PLS for dry soil samples (RMSEP = 0.65 % and RPD = 1.68) and soil samples with 10 % MC (RMSEP = 0.61 % and RPD = 1.71). Whereas the ANN showed better performance for SOC prediction at all MC levels, PLS showed better predictive accuracy of pH at all MC levels except for 25 % MC. Therefore, based on the data set used in the current study, the ANN is recommended for the analyses of SOC at all MC levels, whereas PLS is recommended for the analysis of pH at MC levels below 20 %.
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O método do pH SMP, em virtude de sua praticidade analítica, tem sido amplamente utilizado na estimativa da acidez potencial (H+Al), em substituição à solução de acetato de cálcio 0,5 mol L-1 a pH 7,0, em laboratórios de análise de solo no Brasil. No entanto, tornam-se imprescindíveis estudos regionalizados para estimar com segurança os valores da acidez potencial. Este trabalho objetivou ajustar uma equação de regressão com boa capacidade preditiva dos valores de H+Al, a partir da medida do pH(SMP), para solos do Estado da Paraíba, visando sua utilização em análises de rotina para estimar a acidez potencial. Foram utilizadas 90 amostras de solo coletadas nas várias regiões do Estado. Essas amostras foram analisadas para pH(H2O), pH(SMP), H+Al e pH do sobrenadante da solução de acetato de cálcio. Foram observadas estreitas correlações quadrática simples do pH de equilíbrio da suspensão de acetato de cálcio com a acidez potencial e linear simples com o pH(SMP), com elevada capacidade preditiva (R2 = 0,93 e R2 = 0,95, respectivamente). Os resultados permitiram concluir que a acidez potencial (H+Al), expressa em cmolc dm-3 pode ser estimada pelo método do pH(SMP), por meio da equação H+Al = 57,108 -13,338** SMP + 0,7637** SMP2 (R2 = 0,87).
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Nanoparticles with pH-sensitive behavior may enhance the success of chemotherapy in many cancers by efficient intracellular drug delivery. Here, we investigated the effect of a bioactive surfactant with pH-sensitive properties on the antitumor activity and intracellular behavior of methotrexate-loaded chitosan nanoparticles (MTX-CS-NPs). NPs were prepared using a modified ionotropic complexation process, in which was included the surfactant derived from Nα,Nε-dioctanoyl lysine with an inorganic lithium counterion. The pH-sensitive behavior of NPs allowed accelerated release of MTX in an acidic medium, as well as membrane-lytic pH-dependent activity, which facilitated the cytosolic delivery of endocytosed materials. Moreover, our results clearly proved that MTX-CSNPs were more active against the tumor HeLa and MCF-7 cell lines than the free drug. The feasibilty of using NPs to target acidic tumor extracellular pH was also shown, as cytotoxicity against cancer cells was greater in a mildly acidic environment. Finally, the combined physicochemical and pH-sensitive properties of NPs generally allowed the entrapped drug to induce greater cell cycle arrest and apoptotic effects. Therefore, our overall results suggest that pH-sensitive MTX-CS-NPs could be potentially useful as a carrier system for tumor and intracellular drug delivery in cancer therapy.
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Many strategies for treating diseases require the delivery of drugs into the cell cytoplasm following internalization within endosomal vesicles. Thus, compounds triggered by low pH to disrupt membranes and release endosomal contents into the cytosol are of particular interest. Here, we report novel cationic lysine-based surfactants (hydrochloride salts of N¿- and N¿-acyl lysine methyl ester) that differ in the position of the positive charge and the length of the alkyl chain. Amino acid-based surfactants could be promising novel biomaterials in drug delivery systems, given their biocompatible properties and low cytotoxic potential. We examined their ability to disrupt the cell membrane in a range of pH values, concentrations and incubation times, using a standard hemolysis assay as a model of endosomal membranes. Furthermore, we addressed the mechanism of surfactant-mediated membrane destabilization, including the effects of each surfactant on erythrocyte morphology as a function of pH. We found that only surfactants with the positive charge on the ¿-amino group of lysine showed pH-sensitive hemolytic activity and improved kinetics within the endosomal pH range, indicating that the positive charge position is critical for pH-responsive behavior. Moreover, our results showed that an increase in the alkyl chain length from 14 to 16 carbon atoms was associated with a lower ability to disrupt cell membranes. Knowledge on modulating surfactant-lipid bilayer interactions may help us to develop more efficient biocompatible amino acid-based drug delivery devices.
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The toxicity and environmental behavior of new pH-sensitive surfactants from lysine are presented. Three different chemical structures are studied: surfactants with one amino acid and one alkyl chain, surfactants with two amino acids on the polar head and one alkyl chain, and gemini surfactants. The pH sensitivity of these compounds can be tuned by modifying their chemical structures. Cytotoxicity has been evaluated using erythrocytes and fibroblast cells. The toxic effects against these cells depend on the hydrophobicity of the molecules as well as their cationic charge density. The effect of hydrophobicity and cationic charge density on toxicity is different for each type of cells. For erythrocytes, the toxicity increases as hydrophobicity and charge density increases. Nevertheless, for fibroblasts cationic charge density affects cytotoxicity in the opposite way: the higher charge density, the lower the toxicity. The effect of the pH on hemolysis has been evaluated in detail. The aquatic toxicity was established using Daphnia magna. All surfactants yielded EC50 values considerably higher than that reported for cationic surfactants based on quaternary ammonium groups. Finally, their biodegradability was evaluated using the CO2 headspace test (ISO 14593). These lysine derivatives showed high levels of biodegradation under aerobic conditions and can be classified as"readily biodegradable compounds".
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ABSTRACT Management of boron fertilization depends on the magnitude of B leaching in the soil profile, which varies proportionally with the concentration of B in the soil solution, which, in turn, decreases as the soil pH increases due to the higher sorption of B on soil solid surfaces. The objective of this study was to quantify the effect of liming and rates of B applied to the soil on B leaching. The experiment was carried out in the laboratory in 2012, and treatments consisted of a factorial combination of two rates of liming (without and with lime to raise the soil pH to 6.0) and five rates of B (0, 10, 20, 50 and 100 mg kg-1, as boric acid). A Typic Rhodudalf was used, containing 790 g kg-1 clay and 23 g kg-1 organic matter; the pH(H2O) was 4.6. Experimental units were composed of PVC leaching columns (0.10 m in diameter) containing 1.42 kg of soil (dry base). Boron was manually mixed with the top 0.15 m of the soil. After that, every seven days for 15 weeks, 300 mL of distilled water were added to the top of each column. In the percolated solution, both the volume and concentration of B were measured. Leaching of B decreased with increased soil pH and, averaged across the B rates applied, was 58 % higher from unlimed (pH 4.6) than from limed (pH 6.6) samples as a result of the increase in B sorption with higher soil pH. In spite of its high vertical mobility, the residual effect of B was high in this oxisol, mainly in the limed samples where 80 % of B applied at the two highest rates remained in the soil, even after 15 water percolations. Total recovery of applied B, including leached B plus B extracted from the soil after all percolations, was less than 50 %, showing that not all sorbed B was quantified by the hot water extraction method.
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Audit report on the City of Van Meter, Iowa for the year ended June 30, 2012