Abstract: In order to investigate the possible existence in biological and poorly crystalline synthetic apatites of local atomic organizations different from that of apatite, resolution-enhanced, Fourier transform infrared spectroscopy studies were carried out on chicken bone, pig enamel, and poorly crystalline synthetic apatites containing carbonate and HPO4 2− groups. The spectra obtained were compared to those of synthetic well crystallized apatites (stoichiometric hydroxyapatite, HPO4 2−-containing apatite, type B carbonate apatite) and nonapatitic calcium phosphates which have been suggested as precursors of the apatitic phase [octacalcium phosphate (OCP), brushite, and β tricalcium phosphate and whitlockite]. The spectra of bone and enamel, as well as poorly crystalline, synthetic apatite in thev 4 PO4 domain, exhibit, in addition to the three apatitic bands, three absorption bands that were shown to be independent of the organic matrix. Two low-wave number bands at 520–530 and 540–550 cm−1 are assigned to HPO4 2−. Reference to known calcium phosphates shows that bands in this domain also exist in HPO4 2−-containing apatite, brushite, and OCP. However, the lack of specific absorption bands prevents a clear identification of these HPO4 2− environments. The third absorption band (610–615 cm−1) is not related to HPO4 2− or OH− ions. It appears to be due to a labile PO4 3− environment which could not be identified with any phosphate environment existing in our reference samples, and thus seems specific of poorly crystalline apatites. Correlation of the variations in band intensities show that 610–615 cm−1 band is related to an absorption band at 560 cm−1 superimposed on an apatite band. All the nonapatitic phosphate environments were shown to decrease during aging of enamel, bone, and synthetic apatites. Moreover, EDTA etching show that the labile PO4 3− environment exhibited a heterogeneous distribution in the insoluble precipitate.

Abstract: The potential of strains of Rhizobium and Bradyrhizobium to solubilize rock phosphate was evaluated in vitro. Almost all organisms tested effectively solubilized rock phosphate and lowered the medium pH. The presence or absence of (NH4)2SO4 made little difference in the solubilization of rock phosphate. Among the strains, Rhizobium leguminosarum biovar viceae BICC635 was the most effective solubilizer. Maximum solubilization of phosphate and acid production was achieved after 3 days of incubation. The strain produced 2-ketogluconic acid in the culture medium, the primary cause of rock phosphate solubilization. Increasing the phosphate status of the medium had little effect on the extent of dissolution of Purulia rock phosphate. Adding calcium as CaCl2, CaCO3 and Ca(OH)2 reduced the phosphate solubilization from phosphate rocks. The results indicated that pH, per se, is of less importance in phosphate solubilization. EDTA increased the extent of rock phosphate solubilization, possibly by chelation of the calcium ions that the solubilization produced.

Abstract: Bioactive glasses are glasses which can adhere to bone through chemical bonding. This paper discusses the bonding to bone of phosphate-free glasses, and the formation of calcium phosphate at the surface of these glasses in vivo. The bone bonding was proved by a push-out test. In the bonding to bone, the glass surface initially transforms into a silica gel. By SEM/EDX analysis it was shown that phosphate ions from the physiological solution penetrate far into the gel. Calcium accumulation occurs only in the presence of phosphate. The results suggest that the initial stage of the calcium phosphate build-up is complexation of phosphate by the silica gel.

Abstract: Biomaterials useful for orthopedic and dental applications is disclosed. These materials have a base portion of calcium carbonate and a surface layer of a synthetic phosphate such as hydroxyapatite. The base portion may be a calcium carbonate structure having three-dimensional interconnected porosity such as may be found in porous skeletal carbonate of marine life, e.g. coral porites skeletal aragonite, or it may be porous or non-porous granules of calcium carbonate. A method for making the biomaterials is also disclosed. The synthetic phosphate surface is made using a hydroconversion reaction with a soluble or solubilized phosphate such as ammonium dibasic phosphate ((NH4)2HPO4).

Abstract: The comparative study of the intumescent formulations (poly propylene-ammonium polyphosphate-pentaerythritol) and (polypropylene- ammonium pyrophosphate-pentaerythritol) shows that the highest fire retar dance property of the second system may be related to the existence of a protective carbonaceous coating stable in the temperature range 300-550°C. Comparatively, the stability of the carbonaceous coating on the first system is only observed between 300 and 420°C. A 31P N.M.R. study shows the presence (in the carbon-based layers) of acidic phosphate species (Lewis acids) which result from the thermal degradation of the additive mixtures. In the particular case of (PP-PPA-PER), these species degrade at 420 ° C to give phosphorus oxide. The study reveals that, with (PP-PY-PER), an additional protective char (stable in the temperature range 420-550°C) is formed by a reaction between the Lewis acids and the oxidized products of the thermo-oxidative degradation of the polymer.

Abstract: Suspension-cultured cells of tomato (Lycopersicon esculentum) start to secrete an RNA-degrading enzyme activity during transition from logarithmic to stationary growth phase. Using affinity chromatography on agarose-5-(4-aminophenyl-phosphoryl) uridine 3′(2′) monophosphate as a powerful and final enrichment step, the enzyme was purified to homogeneity and characterized as ribonuclease I (RNase I) according to the following data: (a) it has an Mr of 22,000 (sodium dodecyl sulfate-polyacrylamide gel electrophoresis), a pH-optimum of pH 5.5, a pl of 3.9, and its activity was found to be insensitive to EDTA; (b) the enzyme splits single-stranded RNA endonucleolytically by a phosphotransferase reaction yielding 2′,3′-cNMPs as primary monomeric products; (c) as studied with diribonucleoside monophosphates as substrates, the enzyme exhibits a pronounced preference for 5′ purine residues adjacent to the cleavage site. Most interestingly, in vivo synthesis and secretion was found to be induced when tomato cells were specifically starved for phosphate as mineral nutrient. (a) Extracellular enzyme activity increased about tenfold after transfer of phosphate-grown cells into medium lacking only phosphate. Accordingly, this increase in activity was not detectable when cells were constantly supplied with phosphate. (b) Biosynthetically labeling of the extracellular protein with radioactive amino acids was detectable by sodium dodecyl sulfate-polyacrylamide gel electrophoresis/fluorography directly within the bulk of extracellular proteins. Therefore, we propose that the secreted tomato RNase I synthesized upon phosphate starvation is a component of a higher plant inducible rescue system for scavenging exogenous phosphate.

Abstract: Bean (Phaseolus vulgaris L.) seedlings were cultured on complete or phosphate-deficient nutrient medium. After 14 days of culture on phosphate-deficient medium the visible symptoms of P(i) deficiency were observed only in the shoot, the fresh and dry weights of the roots were slightly higher than in control plants. The decreased P(i) content in the roots had little effect on total respiration rate but had an effect on the level of inhibition of respiration by cyanide. The high resistance of respiration to cyanide observed in P(i) -deficient roots was the result of the suppression of cytochrome path activity and an increased participation of the alternative, cyanide-resistant pathway. The cytochrome pathway activity increased when inorganic phosphate was supplied to P(i) -deficient roots for 1 or 3.5 h. It is speculated that the suppression of cytochrome pathway in P(i) -deficient roots may result from restriction of the phosphorylating capacity or a partial inhibition of cytochrome oxidase activity.

Abstract: The vacuolar and cytoplasmic inorganic phosphate (Pi) content of the mature regions of maize roots was measured by a 31P NMR technique which used an external standard to avoid the need for tissue extraction and which exploited the relatively rapid relaxation of cytoplasmic Pi in order to improve the detection of this pool in fully-vacuolated cells.In mature roots of maize growing with abundant external phosphate, the concentration of Pi in the cytoplasm was approximately 6.5 mol m-3. When these plants were deprived of external phosphate, the vacuolar Pi content of the roots decreased rapidly, but the cytoplasmic Pi concentration initially remained constant and did not begin to decline until P-stress became severe. Calculations show that withdrawal of Pi from the vacuoles into the cytoplasm under these conditions would be against an electrochemical gradient.During P-starvation, an increased capacity for Pi influx developed, preceding any detectable change in the cytoplasmic Pi content of the roots. This response is considered in terms of parallel effects on transport sites for phosphate at the plasmalemma and at the tonoplast. Comparisons of simultaneous rates of influx and net uptake implied that phosphate efflux accounted for < 10% of influx in plants of a steady or declining P-status. However, direct measurements of efflux suggested that this process may be temporarily accelerated when plants are recovering from P-stress. © 1990 Oxford University Press.

Abstract: Upon absorption of phosphate, cultured cells of Catharanthus roseus (L.) G. Don caused a rapid alkalinization of the medium in which they were suspended. The alkalinization continued until the added phosphate was completely exhausted from the medium, at which time the pH of the medium started to drop sharply toward the original pH value. Phosphate exposure caused the pH of the medium to increase from pH 3.5 to values as high as 5.8, while the rate of phosphate uptake was constant throughout (10-17 micromoles per hour per gram fresh weight). This indicates that no apparent pH optimum exists for the phosphate uptake by the cultured cells. The amount of protons cotransported with phosphate was calculated from the observed pH change up to the maximum alkalinization and the titration curve of the cell suspension. Proton/phosphate transport stoichiometry ranged from less than unity to 4 according to the amount of phosphate applied. At low phosphate doses, the stoichiometries were close to 4, while at high phosphate doses, smaller stoichiometries were observed. This suggests that, at high phosphate doses, activation of the proton pump is induced by the longer lasting proton influx acidifying the cytoplasm. The increased H(+) efflux due to the proton pump could partially compensate protons taken up via the proton-phosphate cotransport system. Thus, the H(+)/H(2)PO(4) (-) stoichiometry of the cotransport is most likely to be 4.

Abstract: The iron core of proteins in the ferritin family displays structural variations that include phosphate content as well as the number and the degree of ordering of the iron atoms. Earlier studies had shown that ferritin iron cores naturally high in phosphate, e.g., Azotobacter vinelandii (AV) ferritin (Fe:P ratio = 1:1.7), had decreased long-range order. Here, the influence of phosphate on the local structure around iron in ferritin cores is reported, comparing the EXAFS of AV ferritin, reconstituted ferritin [the protein coats of horse spleen ferritin mixed with Fe(II) with and without phosphate at pH 7] (Fe:P ratio = 1:0.25), and native horse spleen ferritin (Fe:P ratio = 1:0.125); reconstituted horse spleen ferritin without phosphate was indistinguishable from native horse spleen ferritin (HSF) in the analysis. In contrast, when the phosphate content was high in AV ferritin and horse spleen ferritin reconstituted with phosphate, the average iron atom had five to six phosphorus neighbors at 3.17 A. Moreover, the number of detectable iron neighbors was lower when phosphate was high or present during reconstitution (2-3 vs 5-6), and the interatomic distance was longer (3.50 vs 3.03 A), indicating that some phosphate bridges neighboring iron atoms. However, the decrease in the number of detectable iron-iron neighbors compared to HSF and the higher number of Fe-P interactions relative to Fe-Fe interactions suggest that some phosphate ligands were chain termini, or blocked crystal growth, and/or introduced defects which contributed both to the long-range disorder and to altered redox properties previously observed in AV ferritin.

Abstract: The surface reactions of calcium phosphate ceramics have been thought to play an important role in bonding with living bone. Hydroxyapatite (HA), tricalcium phosphate (TCP), and two kinds of apatite-containing glass ceramics were immersed in three types of solutions with different chemical constituents. The first solution was a physiological saline, the second contained phosphate (PO4), and the third was a balanced salt solution consisting of calcium (Ca), magnesium (Mg), and PO4. After serial incubation periods, changes in the solutions were assessed by measurement of total Ca, Mg, and PO4. The ceramic surfaces were studied using scanning electron spectroscopy, infrared reflection spectroscopy, and thin-film x-ray diffraction. The surface reactions of the ceramics were greatly affected by the chemical compositions of the surrounding media. In the complete solution with both Ca and PO4, a carbonated apatite layer was formed on the surfaces of HA, TCP, and the glass ceramics. In comparison to HA and TCP, the glass ceramics were characterized as Ca-releasing materials, the dissolved Ca creating an apatite layer on the surfaces in a few days, in conjunction with PO4 stock in the surrounding media. The immersion test with various solutions proved to be a simple and effective method of assessing surface conditions of ceramic materials.

Abstract: The value of iron ore is adversely affected by phosphorus in concentrations over 0.03% by weight. The present research concerns the use of metabolic products of aPenicillium-like fungus to leach insoluble phosphates (hydroxyapatite) from ores. Ion chromatography was used to measure metabolism of glucose into acidic fragments. The rate and products of glucose degradation depended on both the chemical composition of the growth medium (buffered or not) and incubation conditions (shaken or quiescent). The principal products were identified as oxalic acid and isomers of propylene dicarboxylic acid, mainly itaconic acid. Continued, slow metabolism of itaconic acid generates more oxalic acid. Aliphatic acids were not detected. Both iron ore phosphate and calcium phosphate were partially solubilized by either the spent broth or aqueous oxalic acid. Solubilization of ore phosphorus was greatly assisted by hydrochloric acid added to the spent broth in small increments. The data suggest biological alternatives to costly leaching procedures that use only mineral acids.

Abstract: The effect of soil pH on the dissolution of phosphate rocks (PRs) and the subsequent availability of the dissolved inorganic phosphorus (Pi) to plants was examined in a volcanic soil adjusted to different pH values. Potassium dihydrogen orthophosphate (KH2PO4) and three PRs, Nauru (NPR), Jordan (JPR) and North Carolina (NCPR) were incubated with the pH-amended soils at a rate of 800µg P g−1 soil for 84 days. The extent of PR dissolution was determined by measuring the increases in the amount of 0.5 M NaOH extractable Pi (ΔNaOH-P) in the PR treated soil over the control soil. The amount of plant available P was measured either by extracting with 0.5 M NaHCO3 or by growing ryegrass in soil samples incubated with the phosphate sources. At each pH the order of the extent of PR dissolution followed NCPR > JPR > NPR, which was consistent with the decreasing order of their chemical reactivities. As the pH decreased from 6.5 to 3.9 the dissolution of PRs increased from 29.3% to 83.5%, from 18.2% to 78.9%, and from 12.5% to 60.3% for NCPR, JPR and NPR, respectively. In contrast, as the soil pH decreased from 6.5 to 3.9, the proportion of the dissolved P extracted by 0.5 M NAHCO3 decreased from 38% to 5% and the proportion taken up by ryegrass plants decreased from 46% to 7%. This decrease in plant available P corresponded to an increase in the adsorption of inorganic P with a decrease in pH. However, the uptake of P from PR relative to that from KH2PO4 was higher at low pH than at high pH. Further, the amount of P taken up by plants was more closely related to the amount of NaHCO3 extractable P than to the amount of dissolved P present in the soil.

Abstract: SUMMARY The sorption of phosphate (P) by four strongly acid Fijian soils from 0.01 M CaCl2 decreased with increasing pH up to pH 5.5–6.0 and then increased again. The initial decrease in P sorption with increasing pH appears to result from an interaction between added P, negative charge, and the electrostatic potential in the plane of sorption. The results of a sorption study, involving KCl or CaCl2 of varying concentrations as the background electrolyte and using Nadroloulou soil incubated with KOH or Ca(OH)2, suggested that the increase in P sorption at pH values > 6.0 was caused by the formation of insoluble Ca-P compounds. For some soils this is consistent with the results of an isotopic-exchange study in which incubation with lime caused marked reductions in the amounts of exchangeable P at high pH.

Abstract: Experiments were carried out to determine whether pyrophosphate: fructose-6-phosphate phosphotransferase (PFP) catalyses the rapid recycling of triose phosphates that is found in the cytosol of heterotrophic cell cultures of Chenopodium rubrum L. (W.-D. Hatzfeld, M. Stitt, 1990, Planta, 180, 198–204). Oxygen uptake, carbohydrate turnover, fructose 2,6-bisphosphate (Fru2,6bisP), glycolytic intermediates, adenine and uridine nucleotides, pyrophosphate and the activity of PFP and glycolytic enzymes were monitored for 48 h after subculturing carbohydrate-depleted cells onto glucose. Immediately after transfer there was an increase in the amount of Fru2,6bisP, and of the hexose phosphate. The triose phosphates, fructose-1,6-bisphosphate and inorganic pyrophosphate increased gradually over the next 24 h. This was accompanied by a tripling in the extractable activity of PFP, but not of phosphofructokinase. The activity of fructose-1,6-bisphosphatase was 20–50fold lower than that of PFP. It is calculated that the activity of PFP is high enough to catalyse the observed rate of cycling between the triose phosphates and the hexose phosphates, based on the measured Vmax capacity of the enzyme, the known kinetic properties, and the measured levels of its reactants and Fru2,6bisP. The changes in the levels of Fru2,6bisP were not correlated with the rate of respiration. Instead, the rate of O2 uptake was inversely related to the phosphoenolpyruvate level, showing that pyruvate kinase or phosphoenolpyruvate carboxylase are regulating the use of glucose for respiration. There was also no relation between Fru2,6bisP, and partitioning to sucrose or starch. It is proposed that the main function of the cycle in these cells is to maintain high levels of inorganic pyrophosphate and triose phosphates, which are necessary for the remobilisation of sucrose and for biosynthesis in the plastid, and that ‘coarse’ and ‘fine’ control of PFP play an important role in regulating this cycle.