Physiological and genetic characterization of calcium phosphate precipitation by Pseudomonas species.
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ABSTRACT:
SUBMITTER: Fishman MR
PROVIDER: S-EPMC6033914 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Early studies have shown that moderate levels of calcium overload can cause lower oxidative phosphorylation rates. However, the mechanistic interpretations of these findings were inadequate. And while the effect of excessive calcium overload on mitochondrial function is well appreciated, there has been little to no reports on the consequences of low to moderate calcium overload. To resolve this inadequacy, mitochondrial function from guinea pig hearts was quantified using several well-established methods including high-resolution respirometry and spectrofluorimetry and analyzed using mathematical modeling. We measured key mitochondrial variables such as respiration, mitochondrial membrane potential, buffer calcium, and substrate effects for a range of mitochondrial calcium loads from near zero to levels approaching mitochondrial permeability transition. In addition, we developed a computer model closely mimicking the experimental conditions and used this model to design experiments capable of eliminating many hypotheses generated from the data analysis. We subsequently performed those experiments and determined why mitochondrial ADP-stimulated respiration is significantly lowered during calcium overload. We found that when calcium phosphate levels, not matrix free calcium, reached sufficient levels, complex I activity is inhibited, and the rate of ATP synthesis is reduced. Our findings suggest that calcium phosphate granules form physical barriers that isolate complex I from NADH, disrupt complex I activity, or destabilize cristae and inhibit NADH-dependent respiration.
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Project description:Phosphorus (P) is an essential nutrient for living organisms and cannot be replaced or substituted. In this paper, we present a simple yet efficient membrane free electrochemical system for P removal and recovery as calcium phosphate (CaP). This method relies on in situ formation of hydroxide ions by electro mediated water reduction at a titanium cathode surface. The in situ raised pH at the cathode provides a local environment where CaP will become highly supersaturated. Therefore, homogeneous and heterogeneous nucleation of CaP occurs near and at the cathode surface. Because of the local high pH, the P removal behavior is not sensitive to bulk solution pH and therefore, efficient P removal was observed in three studied bulk solutions with pH of 4.0 (56.1%), 8.2 (57.4%), and 10.0 (48.4%) after 24 h of reaction time. While P removal efficiencies are not generally affected by bulk solution pH, the chemical-physical properties of CaP solids collected on the cathode are still related to bulk solution pH, as confirmed by structure characterizations. High initial solution pH promotes the formation of more crystalline products with relatively high Ca/P molar ratio. The Ca/P molar ratio increases from 1.30 (pH 4.0) to 1.38 (pH 8.2) and further increases to 1.55 (pH 10.0). The formation of CaP precipitates was a typical crystallization process, with an amorphous phase formed at the initial stage which then transforms to the most stable crystal phase, hydroxyapatite, which is inferred from the increased Ca/P molar ratio from 1.38 (day 1) to the theoretical 1.76 (day 11) and by the formation of needle-like crystals. Finally, we demonstrated the efficiency of this system for real wastewater. This, together with the fact that the electrochemical method can work at low bulk pH, without dosing chemicals and a need for a separation process, highlights the potential application of the electrochemical method for P removal and recovery.
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Project description:Calcium phosphates (CaPs) composites with silver nanoparticles (AgNPs) attract attention as a possible alternative to conventional approaches to combating orthopedic implant-associated infections. Although precipitation of calcium phosphates at room temperatures was pointed out as an advantageous method for the preparation of various CaP-based biomaterials, to the best of our knowledge, no such study exists for the preparation of CaPs/AgNP composites. Motivated by this lack of data in this study we investigated the influence of AgNPs stabilized with citrate (cit-AgNPs), poly(vinylpyrrolidone) (PVP-AgNPs), and sodium bis(2-ethylhexyl) sulfosuccinate (AOT-AgNPs) in the concentration range 5–25 mg dm−3 on the precipitation of CaPs. The first solid phase to precipitate in the investigated precipitation system was amorphous calcium phosphate (ACP). The effect of AgNPs on ACP stability was significant only in the presence of the highest concentration of AOT-AgNPs. However, in all precipitation systems containing AgNPs, the morphology of ACP was affected, as gel-like precipitates formed in addition to the typical chain-like aggregates of spherical particles. The exact effect depended on the type of AgNPs. After 60 min of reaction time, a mixture of calcium-deficient hydroxyapatite (CaDHA) and a smaller amount of octacalcium phosphate (OCP) formed. PXRD and EPR data point out that the amount of formed OCP decreases with increasing AgNPs concentration. The obtained results showed that AgNPs can modify the precipitation of CaPs and that CaPs properties can be fine-tuned by the choice of stabilizing agent. Furthermore, it was shown that precipitation can be used as a simple and fast method for CaP/AgNPs composites preparation which is of special interest for biomaterials preparation.
| S-EPMC10003846 | biostudies-literature
Project description:Calcium (Ca) is an essential plant nutrient, required for signaling, cell wall fortification and growth and development. Calcium deficiency (Ca-deficiency) in maize causes leaf tip rot and a so-called "bull-whipping" or "buggy-whipping" phenotype. Seedlings of the maize line B73 displayed these Ca-deficiency-like symptoms when grown in the greenhouse with excess fertilizer during the winter months, while seedlings of the Mo17 maize line did not display these symptoms under the same conditions. These differential phenotypes could be recapitulated in 'mini-hydroponic' systems in the laboratory in which high ammonium, but not nitrate, levels induced the symptoms in B73 but not Mo17 seedlings. Consistent with this phenotype being caused by Ca-deficiency, addition of Ca2+ completely relieved the symptoms. These data suggest that ammonium reduces the seedling's ability to absorb calcium, which causes the Ca-deficiency phenotype, and that this trait varies among genotypes. A recombinant inbred line (RIL) population derived from a B73 x Mo17 cross was used to map quantitative trait loci (QTL) associated with the Ca-deficiency phenotype. QTL associated with variation in susceptibility to Ca-deficiency were detected on chromosomes 1, 2, 3, 6 which explained between 3.30-9.94% of the observed variation. Several genes predicted to bind or be activated by calcium map to these QTL on chromosome 1, 2, 6. These results describe for the first time the genetics of Ca-deficiency symptoms in maize and in plants in general.
| S-EPMC7263677 | biostudies-literature
Project description:Alzheimer's disease (AD), the most common form of dementia, is manifested in the brain by the aggregation of amyloid plaques and neurofibrillary tangles. The tangles are primarily composed of microtubule-associated protein tau that is aberrantly hyperphosphorylated, suggesting that deregulated phosphorylation may contribute to AD pathogenesis. However, systematic analysis of the phosphoproteome in AD brain tissues has not been reported. We used calcium phosphate precipitation to analyze an AD postmortem brain, followed by liquid chromatography-tandem mass spectrometry. The protein sample was first resolved by one-dimensional polyacrylamide gel electrophoresis and subjected to gel excision and in-gel digestion. Phosphopeptides in the resulting peptide mixtures were enriched in a single step of calcium phosphate precipitation, and then analyzed by the LC-MS/MS approach. After database search, stringent filtering, and manual validation of neutral loss in the MS/MS spectra, a total of 466 phosphorylation sites on 185 proteins including tau were identified. A majority of sites were not described previously. This study demonstrates the feasibility of combining calcium phosphate precipitation with mass spectrometry for phosphoproteome analysis of postmortem human brain tissue.
| S-EPMC2663965 | biostudies-literature
Project description:The ability of matrix vesicles isolated from the epiphysial growth plate of 6-week-old chicks to facilitate the precipitation of calcium phosphate was studied in vitro. The vesicles lowered the minimum concentration product [ca2+]X[p1] needed to induce crystal formation, thereby showing the vesicles are nucleators of crystallization. After freezing and thawing the vesicles at pH6.0, part but not all of this ability to nucleate disappeared. Freezing and thawing markedly decreased the Ca and Pi content of the vesicles, suggesting that part of the nucleating activity may have been due to mineral already present. After removal of the mineral the residual nucleating activity could be destroyed by extracting the vesicles with lipid solvents or by treatment with enzymes such as phosphoilipase C, neuraminidase or proteinase. Matrix vesicles obtained from chicks treated with 1-hydroxyethane-1, 1-diphosphonate, a compound that inhibits calcification in vivo, showed impaired nucleating activity, both before and after treatment at pH6.0. The vesicle preparation bound some diphosphonate in vitro, probably to the mineral present in the preparation, since no binding could be detected in vesicles preincubated at pH6.0. No difference was found in the nucleating activity of vesicles isolated from rachitic chicks which had or had not received cholacalciferol 48 h before death. These results suggest that matrix vesicles possess intrinsic nucleating activity that may be important in biological calcification.
| S-EPMC1183947 | biostudies-other
Project description:Phosphorus (P) is a vital micronutrient element for all life forms. Typically, P can be extracted from phosphate rock. Unfortunately, the phosphate rock is a nonrenewable resource with a limited reserve on the earth. High levels of P discharged to water bodies lead to eutrophication. Therefore, P needs to be removed and is preferably recovered as an additional P source. A possible way to achieve this goal is by electrochemically induced phosphate precipitation with coexisting calcium ions. Here, we report a new concept of phosphate removal and recovery, namely a CaCO3 packed electrochemical precipitation column, which achieved improved removal efficiency, shortened hydraulic retention time, and substantially enhanced stability, compared with our previous electrochemical system. The concept is based on the introduction of CaCO3 particles, which facilitates calcium phosphate precipitation by buffering the formed H+ at the anode, releases Ca2+, acts as seeds, and establishes a high pH environment in the bulk solution in addition to that in the vicinity of the cathode. It was found that the applied current, the CaCO3 particle size, and the feed rate affect the removal of phosphate. Under optimized conditions (particle size, <0.5 mm; feed rate, 0.4 L/d; current, 5 mA), in a continuous flow system, the CaCO3 packed electrochemical precipitation column achieved 90 ± 5% removal of phosphate in 40 days and >50% removal over 125 days with little maintenance. The specific energy consumptions of this system lie between 29 and 61 kWh/kg P. The experimental results demonstrate the promising potential of the CaCO3 packed electrochemical precipitation column for P removal and recovery from P-containing streams.
| S-EPMC6876926 | biostudies-literature
Project description:Amelogenin is essential for proper enamel formation. The present in vitro study extends our previous work at low (10 mM) ionic strength (IS) by examining the effect of amelogenin on mineralization under higher (162 mM) IS conditions found in developing enamel. Full-length phosphorylated (P173) and non-phosphorylated (rP172) amelogenins were examined, along with P148 and rP147 that lack the hydrophilic C-terminus. Calcium phosphate formation was assessed by pH change, while the minerals formed were characterized using transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy. Amelogenin self-assembly was also studied using dynamic light scattering and TEM. The results indicate that IS does not influence the effects of rP147, rP172, and P173 on mineralization. However, in contrast to the findings for low IS, where both P173 and P148 stabilize initially formed amorphous calcium phosphate (ACP) nanoparticles for >1 d, elongated hydroxyapatite crystals were observed after 24 h using P148 at high IS, unlike that seen with P173. Differences in self-assembly help explain these findings, which suggest that P173 and P148 may play different roles in regulating enamel mineral formation. The present data support the notion that proteolytic processing of P173 is required in vivo to induce the transformation of initial ACP phases to apatitic enamel crystals.
| S-EPMC3448280 | biostudies-literature
Project description:Many questions remain regarding the formation of ultrathin hydroxapatite (HAP) crystals within the confines of collagen fibrils of bones. These structures form through the interplay of the collagen matrix and non-collagenous proteins, and in vitro mineralization studies employing poly(aspartic acid) (PAsp) as a mimic of the non-collagenous proteins have generated mineralized fibrils with structures comparable to their biogenic counterparts. In this article, we employ the nanoscale cylindrical pores perforating track-etch filtration membranes to investigate the role of PAsp in controlling the infiltration and crystallization of calcium phosphate (CaP) within confined volumes. Oriented polycrystalline HAP and non-oriented octacalcium phosphate (OCP) rods precipitated within the membrane pores via an amorphous calcium phosphate (ACP) precursor, where PAsp increased the proportion of OCP rods. Further, ACP crystallized faster within the membranes than in bulk solution when PAsp was present, suggesting that PAsp inhibits crystallization in solution, but promotes it when bound to a substrate. Finally, in contrast to the collagen system, PAsp reduced the yield of intra-membrane mineral and failed to enhance infiltration. This suggests that a specific interaction between the collagen matrix and ACP/PAsp precursor particles drives effective infiltration. Thus, while orientation of HAP crystals can be achieved by confinement alone, the chemistry of the collagen matrix is necessary for efficient mineralisation with CaP.
| S-EPMC3881609 | biostudies-literature
Project description:This work describes a simple sample pretreatment method for the fractionation of human milk proteins (into their two main groups: whey and caseins) prior to their analysis. The protein extraction protocol is based on the addition of calcium phosphate to non-adjusted pH human milk. The combination of calcium ions with phosphate results in an effective co-precipitation of caseins. To assess the suitability of this fractionation protocol, the protein extracts were analyzed by SDS-PAGE, LC-MS/MS and CE analysis. The results evidence a significant decrease in contamination of casein fraction with whey proteins and vice versa compared with the conventional isoelectric precipitation of caseins. In addition, CE fraction collection coupled to LC-MS/MS (off-line coupling) has been successfully applied to the identification of minor proteins in this complex matrix. The methodology presented here constitutes a promising tool to enlarge the knowledge of human milk proteome.
2018-08-31 | PXD010315 | Pride