Abstract: The salt of milk constitutes a small part of milk (8-9 g.L(-1)); this fraction contains calcium, magnesium, sodium and potassium for the main cations and inorganic phosphate, citrate and chloride for the main anions. In milk, these ions are more or less associated between themselves and with proteins. Depending on the type of ion, they are diffusible (cases of sodium, potassium and chloride) or partially associated with casein molecules (cases of calcium, magnesium, phosphate and citrate), to form large colloidal particles called casein micelles. Today, our knowledge and understanding concerning this fraction is relatively complete. In this review, the different models explaining (i) the nature and distribution of these minerals (especially calcium phosphate) in both fractions of milk and (ii) their behaviour in different physico-chemical conditions, are discussed.

Abstract: The renal handling of inorganic phosphate (Pi) is controlled not only by PTH, but also by hitherto undetermined mechanisms dependent on phosphate intake. Recently, fibroblast growth factor (FGF)-23 was identified as a novel phosphaturic factor in tumor-induced osteomalacia and autosomal-dominant hypophosphatemic rickets. We hypothesized that phosphate intake could influence FGF-23 concomitantly to the changes in renal Pi handling. Twenty-nine healthy males were subjected to a 5-d low-phosphate diet and a phosphate binder, followed by a high-phosphate diet including supplements. Concomitant modifications in calcium intake allowed minimizing PTH changes in response to dietary phosphate. Serum FGF-23 levels significantly decreased on the low-phosphate diet, then increased with the oral phosphate load. Changes in FGF-23 were positively correlated with changes in 24-h urinary Pi excretion and negatively correlated with changes in the maximal tubular reabsorption of Pi and 1,25(OH)(2)D(3) (calcitriol), whereas PTH was not. In multivariate analysis, changes in FGF-23 remained the most significantly correlated to changes in 1,25(OH)(2)D(3) and maximal tubular reabsorption of Pi. Moreover, FGF-23 was positively correlated to serum osteocalcin, a marker of osteoblastic activity. In summary, FGF-23 was inversely related to renal Pi transport and serum calcitriol levels in healthy young men. These data suggest that FGF-23 may be implicated in the physiological regulation of Pi homeostasis in response to dietary phosphate changes, independent of PTH.

Abstract: In the present work, the surface of chitosan membranes was modified using a phosphorylation method carried out at room temperature. Phosphorylation may be of particular interest in materials for orthopaedic applications, due to the cation-exchange properties of phosphate functionalities. Phosphate groups chelate calcium ions, thus inducing the deposition of an apatite-like layer known to improve the osteoconduction of polymer-based implants. Additionally, the negatively charged phosphate functionalities, together with the positively charged amine groups from chitosan, are expected to provide chitosan with an amphoteric character, which may be useful as a combinatorial therapeutic strategy, by simultaneously allowing the immobilization of signalling molecules like growth factors. Phosphorylation was carried out at room temperature using the H3PO4/Et3PO4/P2O5/butanol method. Surface characterization was performed by XPS, ATR-FT-IR, and SEM. Cross-sections were analyzed by SEM fitted with EDS. The phosphate content increased with the reaction time, as shown by XPS and ATR-FT-IR, a P/N atomic ratio of 0.73 being obtained after 48 h of treatment. High-resolution XPS spectra regarding C1s, O1s, N1s and P2p are discussed. The introduction of a neutralization step led to a reduction of P content, which pointed out to the presence of phosphates ionically bound to protonated amines, in addition to phosphate esters. EDS analysis of cross-sections revealed a gradual P reduction up to 50% towards the inner part of the membrane.

Abstract: A new route for preparing hydroxyapatite (Ca 10 (PO 4 ) 6 (OH) 2 ) bioceramic has been described. An amorphous, nanosized, and carbonate-containing calcium phosphate powder that had a Ca:P ratio of 1.67 was synthesized from calcium diethoxide and phosphoric acid in ethanol via a sol-gel method. The powder was pressed at 98 MPa into green specimens and then heated to a temperature range of 500°-1300°C. At 600°C, the powder crystallized to a carbonated hydroxyapatite and a trace of β-tricalcium phosphate before converting to hydroxyapatite at 900°C. The thermal crystallization was associated with grain growth, shrinkage, and active surface diffusion. The activation energy of grain growth was 37 ± 2 kJ/mol. After sintering at 1100°C, the decomposition of carbonated hydroxyapatite generated a microporous ceramic with an average pore size of 0.2 μm and an open porosity of 15.5%. This microporous bioceramic can be used as a bone filler.

Abstract: Being a major cause of eutrophication and subsequent loss of water quality, the turnover of phosphorus (P) in lake sediments is in need of deeper understanding. A major part of the flux of P to eutrophic lake sediments is organically bound or of biogenic origin. This P is incorporated in a poorly described mixture of autochthonous and allochthonous sediment and forms the primary storage of P available for recycling to the water column, thus regulating lake trophic status. To identify and quantify biogenic sediment P and assess its lability, we analyzed sediment cores from Lake Erken, Sweden, using traditional P fractionation, and in parallel, NaOH extracts were analyzed using 31P NMR. The surface sediments contain orthophosphates (ortho-P) and pyrophosphates (pyro-P), as well as phosphate mono- and diesters. The first group of compounds to disappear with increased sediment depth is pyrophosphate, followed by a steady decline of the different ester compounds. Estimated half-life times of these compound groups are about 10 yr for pyrophosphate and 2 decades for mono- and diesters. Probably, these compounds will be mineralized to ortho-P and is thus potentially available for recycling to the water column, supporting further growth of phytoplankton. In conclusion, 31P NMR is a useful tool to asses the bioavailability of certain P compound groups, and the combination with traditional fractionation techniques makes quantification possible.

Abstract: Geochemical cycling of phosphorus (P) in aquatic environments is carried out almost exclusively by biota and involves reactions that are catalyzed by enzymes. Oxygen isotope effects accompanying phosphoenzymatic reactions have been determined in controlled laboratory experiments in order to elucidate processes underlying biogeochemical cycling of P, and to identify possible reaction pathways for P-compounds in nature. Phosphate oxygen isotope effects are distinct for specific enzymatic reaction mechanisms measured in microbial culture experiments and in cell-free systems. P 16 O4 is taken up preferentially from inorganic phosphate (Pi )i n the growth medium by intact E. coli cells, producing a kinetic fractionation in the extracellular dissolved Pi pool. Inorganic pyrophosphatase is the intracellular enzyme that catalyzes the temperature-dependent equilibrium oxygen isotope fractionations between phosphates and water in biological systems, and imprints an equilibrium isotope signature on Pi that is turned over or cycled by intact cells. Alkaline phospha- tase, a key enzyme involved in extracellular Pi regeneration in aquatic systems, catalyzes hydrolysis of phosphomonoesters, reactions that are accompanied by kinetic fractionations and disequilibrium (inheritance) isotope effects in released Pi. Compari- son of laboratory determined enzyme-specific isotopic fractionations with those ob- served in microbial culture experiments and in natural aquatic systems, provide new insights into processes controlling P cycling and the relations between P availability and the cycling of N and C. Isotopic signatures associated with specific cellular processes and phosphoenzyme reaction pathways may be useful in assessing P status and for tracing P turnover.

Abstract: Plants inoculated with arbuscular mycorrhizal (AM) fungi utilize more soluble phosphorus from soil mineral phosphate than non-inoculated plants. However, there is no information on the response of soil microflora to mineral phosphate weathering by AM fungi and, in particular, on the catabolic diversity of soil microbial communities. The AM fungus, Glomus intraradices was examined for (i) its effect on the growth of Acacia holosericea, (ii) plant-available phosphate and (iii) soil microbial activity with and without added rock phosphate. After 4-months culture, AM fungal inoculation significantly increased the plant biomasses (by 1.78× and 2.23× for shoot and root biomasses, respectively), while mineral phosphate amendment had no effect in a sterilized soil. After 12-months culture, the biomasses of A. holosericea plants growing in a non-sterilized soil amended with mineral phosphate were significantly higher than those recorded in the control treatment (by 2.5× and 5× for shoot and root biomasses, respectively). The fungal inoculation also significantly stimulated plant growth, which was significantly higher than that measured in the mineral phosphate treatment. When G. intraradices and mineral phosphate were added together to the soil, shoot growth were significantly stimulated over the single treatments (inoculation or amendment) (1.45×). The P leaf mineral content was also higher in the G. intraradices+mineral phosphate treatment than in G. intraradices or rock phosphate amendment. Moreover, the number of fluorescent pseudomonads has been significantly increased when G. intraradices and/or mineral phosphate were added to the soil. By using a specific type of multivariate analysis (co-inertia analysis), it has been shown that plant growth was positively correlated to the metabolization of ketoglutaric acid, and negatively linked to the metabolisation of phenylalanine and other substrates, which shows that microbial activity is also affected. G. intraradices inoculation is highly beneficial to the growth of A. holosericea plants in controlled conditions. This AM symbiosis optimises the P solubilization from the mineral phosphate and affects microbial activity in the hyphosphere of A. holosericea plants.

Abstract: The effects of high aqueous carbonate concentrations on arsenic mobility and transport in the subsurface were studied in synthetic iron oxide-coated sand column experiments. Elevated aqueous carbonate concentrations in groundwater have been studied and linked, by some authors, to increased aqueous As concentrations in natural waters. This study found that increasing carbonate concentrations had relatively little effect on As(V) adsorption to the iron oxide-coated sand surface at pH 7. The adsorption of As(V) decreased marginally when the CO2(g) partial pressure increased from 10(-3.5) to 10(-1.8) atm, despite a 50-fold increase in total dissolved carbonate (0.072 to 3.58 mM). Increasing the CO2(g) partial pressure to 10(-10) atm resulted in only a slight decrease in As(V) adsorption and increase in mobility, despite a >300-fold increase in total dissolved carbonate (to 22.7 mM). When compared to phosphate, a known competitive anion, carbonate mobilized less adsorbed As(V) than was mobilized by phosphate, even when present in much higher concentrations than phosphate. This was also true for an experiment with lower pore water velocity and an experiment where As(II) was introduced instead of As(V). Our experiments conclude that while carbonate anions do compete with As for adsorption to iron oxide-coated sand, the competitive effect is relatively small with regard to the total concentration of adsorbed As and the potential competitive effects of phosphate.

Abstract: Oncogenic osteomalacia (OO), a tumor-associated phosphate-wasting syndrome, provides an opportunity to identify regulators of renal phosphate homeostasis. We established cultures from OO-associated tumors. Conditioned medium from these cultures inhibited phosphate uptake in renal tubular epithelial cells. We then compared RNA from tumor-derived cultures expressing inhibitory activity with RNA from tumor-derived cultures in which inhibitory activity was not evident and identified candidate mRNAs specifically expressed by cultures inhibiting renal phosphate transport. Testing of identified candidates revealed that one protein, fibroblast growth factor 7 (FGF7), was a potent and direct inhibitor of phosphate uptake in vitro. A neutralizing monoclonal antibody to FGF7 reversed FGF7-dependent phosphate transport inhibition and inhibitory activity in conditioned medium from tumor cell cultures. Immunoassay revealed abundant FGF7 in inhibitory conditioned medium and minimal amounts in nonconditioned medium or conditioned medium with no phosphate transport inhibitory activity. Furthermore, only small amounts of FGF23 were present in inhibitory conditioned medium, comparable to concentrations found in conditioned medium with no phosphate transport inhibitory activity. Thus, FGF7 was specifically identified when selecting for in vitro phosphate transport inhibitory activity of tumor-derived cultures and was confirmed as a potent inhibitor of phosphate transport. Finally, FGF7 message was confirmed in PCR products of mRNA extracted from fragments of each tumor. Members of the FGF family (other than FGF23) are expressed by OO-associated tumors and may play a role in mediating this syndrome.

Abstract: Purple acid phosphatases (PAPs) are a family of binuclear metalloenzymes that catalyze the hydrolysis of phosphoric acid esters and anhydrides. A PAP in sweet potato has a unique, strongly antiferromagnetically coupled Fe(III)–Mn(II) center and is distinguished from other PAPs by its increased catalytic efficiency for a range of activated and unactivated phosphate esters, its strict requirement for Mn(II), and the presence of a μ-oxo bridge at pH 4.90. This enzyme displays maximum catalytic efficiency (kcat/Km) at pH 4.5, whereas its catalytic rate constant (kcat) is maximal at near-neutral pH, and, in contrast to other PAPs, its catalytic parameters are not dependent on the pKa of the leaving group. The crystal structure of the phosphate-bound Fe(III)–Mn(II) PAP has been determined to 2.5-A resolution (final Rfree value of 0.256). Structural comparisons of the active site of sweet potato, red kidney bean, and mammalian PAPs show several amino acid substitutions in the sweet potato enzyme that can account for its increased catalytic efficiency. The phosphate molecule binds in an unusual tripodal mode to the two metal ions, with two of the phosphate oxygen atoms binding to Fe(III) and Mn(II), a third oxygen atom bridging the two metal ions, and the fourth oxygen pointing toward the substrate binding pocket. This binding mode is unique among the known structures in this family but is reminiscent of phosphate binding to urease and of sulfate binding to λ protein phosphatase. The structure and kinetics support the hypothesis that the bridging oxygen atom initiates hydrolysis.

Abstract: Phosphorus is recognized as the most critical nutrient limiting lake productivity. The trophic status and development of lake systems are also influenced by the phosphorus content and fractions and phosphate sorption characteristics of the lake sediments. The phosphorus fractions and phosphate sorption characteristics of sediments in shallow lakes from the middle and lower reaches of Yangtze River region in China were investigated. The results show that the phosphorus contents in the sediments ranged from 217.8 to 1640 mg kg(-1); inorganic phosphorus (IP) was the major fraction of total phosphorus (TP); phosphorus bound to Al, Fe, Mn oxides, and hydroxides (Fe/Al-P), and calcium bound phosphorus (Ca-P) were the main fractions of IP. Phosphate sorption on the sediments mainly occurred within 2 h and then reached equilibrium in 10 h. The phosphate sorption rate was closely related to the concentration of fine particles. The phosphate sorption capacity ranged from 128.21 to 833.33 mg kg(-1), showing a significant correlation with the contents of Fe, Fe+Al, total organic carbon (TOC), cationic exchange capacity, total nitrogen, TP, Ca, IP, and the ratio of P/(Al+Fe), and it was higher in the sediments of eutrophic lakes than in mesotrophic lakes. Phosphate was mainly sorbed onto Fe and Al particles. The phosphate sorption efficiency ranged from 26.74 to 312.50 L kg(-1), and had a strong positive correlation with Fe content. For the eutrophic lake sediments, there were no significant relationships between the phosphate sorption efficiency and the selected physical and chemical parameters. But for the mesotrophic lake sediments, the phosphate sorption efficiency was found to be positively related to the contents of Al and Fe+Al.

Abstract: Phosphate deposits are generally characterized by enhanced radionuclide concentrations compared to natural levels. The mining and processing of this phosphate ore redistribute radionuclides throughout the environment and introduce them into phosphoric acid and phosphogypsum. Phosphoric acid is the starting material for triple superphosphate (TSP), single superphosphate (SSP), monoammonium phosphate (MAP), diammonium phosphate (DAP), NPK fertilizers and di-calcium phosphate (DCP). Contents of natural radionuclides from thorium and uranium series, 226 Ra, 210 Pb and 228 Ra, were measured in Brazilian igneous phosphate rock, phosphoric acid, phosphogypsum and phosphate fertilizer samples, using high-resolution gamma-spectrometry. Neutron activation analysis was used for the determination of U and Th in the same samples. The fertilizers samples which are derived directly from phosphoric acid, MAP and DAP, presented activity concentrations around the detection limits of the counting system for 226 Ra (<5.0 Bq . kg –1 ), for 228 Ra (<3.0 Bq . kg –1 ) and for 210 Pb (<19 Bq . kg –1 ). As for U and Th, the concentrations found in MAP and DAP are more significant, up to 374 and 250 Bq . kg –1 , respectively. SSP, TSP and NPK, which are obtained by mixing phosphoric acid with different amounts of phosphate rock and NH3, presented higher concentrations of radionuclides, up to 871 Bq . kg –1 for 226 Ra, 283 Bq . kg –1 for 228 Ra, 1255 Bq . kg –1 for 210 Pb, 413 Bq . kg –1 for U and 538 Bq . kg –1 for Th.

Abstract: Novel calcium phosphate materials were synthesized from solutions containing the surfactant bis(2-ethylhexyl)sulfosuccinate sodium salt (AOT), water and oil. A range of morphologies was obtained by varying the relative concentrations of the solution components. A material with structural features resembling tooth enamel was produced from a highly viscous reaction solution. This material consisted of bundles of co-aligned filaments, 750 nm–1 µm in length and 250–350 nm wide. Each bundle contained 10–20 filaments, identified as hydroxyapatite crystals. Electron diffraction of the bundles resulted in an arc pattern indicative of elongated aligned crystals, and similar to that known for dental enamel. Systems such as this may be used as models to gain insight into the mechanisms involved in the biomineralization of tooth enamel. Importantly, we have provided new evidence in support of the hypothesis that hierarchical structures in nature result from cooperative interactions between organic assembly and crystal growth. Amorphous calcium phosphate nanoparticles, hollow spheres, spherical octacalcium phosphate aggregates of plates and elongated plates of calcium hydrogen phosphate dihydrate were also obtained under different sets of conditions that altered AOT assembly and solution viscosity. These latter findings illustrate further the significant influence of organic assembly on the formation of calcium phosphate materials.

Abstract: Depending on the calcium:phosphorus molar ratio of the initial precipitates determined by precipitation conditions (calcium:phosphorus molar ratio of reactants and pH of reaction environment), after sintering at 1250°C, monophase, biphase, or triphase ceramics consisting of hydroxyapatite, β-tricalcium phosphate, α-tricalcium phosphate, and calcium oxide were obtained. The phase composition and properties—i.e., density, shrinkage, hardness, bending strength, and roughness—of the fractured surfaces of the isostatically re-pressed sinters were determined.

Abstract: To survive in a dynamic and unpredictable environment, cells must correctly interpret and integrate extracellular signals with internal factors. In particular, internal stores of nutrients must be managed for use during periods of nutrient limitation. To gain insight into this complex process, we combined biochemical and spectroscopic techniques to follow the dynamics of the phosphate responsive signaling pathway in both single yeast cells and populations. We demonstrate that the phosphate-responsive genes PHO5 and PHO84 exhibit different kinetics of transcriptional induction in response to phosphate starvation, and that transient phosphate limitation causes induction of PHO84 but not PHO5. This differential kinetic behavior is largely eliminated in cells that lack the ability to store phosphate internally in the form of polyphosphate, but the threshold of external phosphate required for induction of PHO5 and PHO84 is unaffected. Our observations indicate that polyphosphate acts as a buffer that can be mobilized during periods of phosphate limitation and enables the phosphate-responsive signaling pathway to filter transient fluctuations in extracellular phosphate levels.

Abstract: By measuring phosphate uptake by Mycobacterium tuberculosis strains with the pstS1 and pstS2 genes genetically inactivated, we showed that these pstS genes encode high-affinity phosphate binding proteins. In a mouse infection model, both mutants were attenuated in virulence, suggesting that M. tuberculosis encounters limiting phosphate concentrations during its intracellular life span.