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Oxalate ions and calcium oxalate crystals stimulate MCP-1 expression by renal epithelial cells.

BACKGROUND: Crystals of calcium oxalate monohydrate (COM) and excess oxalate ions (OX) stimulate an array of responses inducing localized injury and inflammation in the kidneys. These inflammatory responses are key regulators of development of nephrolithiasis. We propose that monocyte chemoattractant protein-1 (MCP-1), a chemokine with potent chemotactic activity for monocytes/macrophages, is a mediator of local inflammatory responses to COM and OX-induced injury. To test this hypothesis, the effects of COM and OX on the expression of MCP-1 mRNA and protein by NRK52E rat renal tubular cells were investigated. METHODS: Confluent cultures of NRK52E cells were exposed to COM (33 to 267 microg/cm2) or OX (125 to 1000 micromol/L, estimated free oxalate levels of 65.8 to 540 micromol/L) and catalase (400 or 2000 U/mL), a free radical scavenger that protects the cells against detrimental effects of COM and OX, for 1 to 48 hours under serum free conditions. The conditioned media were collected and total cellular RNA isolated from the cells and subjected to enzyme-linked immunosorbent assay (ELISA) and semiquantitative polymerase chain reaction (PCR) to determine the expression of MCP-1 protein and mRNA, respectively. RESULTS: NRK52E cells express MCP-1 mRNA and protein, and the level of their expression significantly increases following treatments with COM and OX in a time and concentration dependent manner. MCP-1 mRNA expression and protein production increased more significantly after exposure to COM than to OX. These responses were significantly reduced following treatments with catalase (2000 U/mL). CONCLUSIONS: NRK52E cells express MCP-1 mRNA and protein, and their levels are altered following COM and OX exposure. Since catalase treatment reduced MCP-1 expression, free radicals may be involved in the up-regulation of MCP-1 production by the epithelial cells. The results suggest that elevated expression of MCP-1, which is often associated with local inflammatory response, may mediate similar reactions including attraction of macrophages seen around the interstitial crystals during the early stages of nephrolithiasis.

Animals↗

Scanning electron microscopic study of calcium oxalate concretions grown in gel system and calcium oxalate stones.

Calcium oxalate concretions grown in gelatin gel medium, and calcium oxalate renal stones were studied by polarized light and scanning electron microscopy. In both cases, the results obtained confirm that the surface crystals have random axial orientation and that the gross configuration seems to be determined by the fibrous organic matrix. In vitro concretions grown in the gelatin gel medium are more resistant to EDTA demineralization and to ultrasonic irradiation than calcium oxalate stones.

Calcium↗

Normal calcium oxalate crystal growth inhibition in severe calcium oxalate nephrolithiasis.

Urine from mammalian kidneys is regularly supersaturated with respect to calcium oxalate monohydrate, the most common solid phase in human nephrolithiasis, and also inhibits the nucleation, growth, and aggregation of calcium oxalate crystals. Nephrolithiasis is often associated with increased supersaturation, and it is assumed that this increase overbalances the inhibition effects, causing stones. However, some patients form stones in the absence of increased supersaturation, and in those patients, one might assume that reduced inhibition is the cause of their stones. This hypothesis was tested in 25 patients who formed at least ten stones each, yet lacked the usual metabolic abnormalities that increase supersaturation. Compared with 25 age- and sex-matched control subjects, urine supersaturation among the patients was not increased; this is an expected result of this study's selection criteria. Compared with the same age- and sex-matched control subjects, urine from the patients showed no evidence for reduced inhibition of calcium oxalate crystal growth, so low inhibition of growth did not contribute to pathogenesis of stones in our highly selected study population, despite their otherwise unexplained and active stone formation. These results do not support the hypothesis that growth inhibition defects are a cause of stone disease.

Calcium Oxalate↗

[Comparison of calcium oxalate values as indicators of risk for calcium oxalate calcinosis and the degree of urine saturation in patients with calcium urolithiasis and in healthy patients].

Daily excretion of calcium, magnesium, oxalates, and citrates together with daily urine output were determined in 37 patients with calcium urolithiasis and in 25 healthy individuals. Basing on the obtained values, a degree of urine saturation with calcium oxalate with Marshall and Robertson technique and a value of risk factor with Tiselius technique were calculated. It was found that daily diuresis and excretion of calcium with the urine are significantly higher in patients with urolithiasis where as daily excretion of citrates with the urine is significantly lower than in healthy individuals. Risk index proved two-fold higher in the examined patients than in the healthy individuals (p greater than 0.001) while the degree of urine saturation with calcium oxalate did not differ significantly in both groups. The authors, discussing causes of seemingly different changes in both tested parameters, stressed diagnostic value of risk index which includes excretion of crystallization inhibitors (magnesium, citrates) and contrary to the degree of urine saturation is independent of daily urine output.

Adult↗

Kinetics and Mechanism of the Nucleophilic Substitution Reaction of Imidazole with Bis(2,4,6-trichlorophenyl) Oxalate and Bis(2,4-dinitrophenyl) Oxalate.

The kinetics of the imidazole-catalyzed decomposition of bis(2,4,6-trichlorophenyl) oxalate (TCPO) and bis(2,4-dinitrophenyl) oxalate (DNPO) was investigated by the stopped-flow technique. Pseudo-first-order rate constants were determined as a function imidazole concentration in the temperature range 6-45 degrees C by fitting the temporal changes in absorbance throughout the 245 to 345 nm wavelength range for TCPO and at 420 nm for DNPO. The reaction proceeds by release of two molecules of substituted phenol and formation of 1,1'-oxalyldiimidazole (ODI) for both esters. The identity of ODI was confirmed in the reaction of imidazole with TCPO by its UV absorbance spectrum and (13)C-NMR spectrum. The reaction of imidazole with TCPO has a second-order dependence on imidazole concentration and an observed negative activation energy of -6.2 +/- 0.3 kJ/mol, whereas the DNPO reaction has a first-order dependence on imidazole concentration and an observed positive activation energy of 12.0 +/- 0.6 kJ/mol. The differences in the temperature dependence and order of the reaction with respect to imidazole for the two oxalate esters are explained by a shift in the rate-determining step from addition to the acyl group for DNPO to imidazole-catalyzed release of the phenol leaving group for TCPO. These kinetics results are useful in interpreting the initial reaction steps in peroxyoxalate chemiluminescence.

Journal Article↗

Madin-Darby canine kidney cells are injured by exposure to oxalate and to calcium oxalate crystals.

The reaction of Madin-Darby canine kidney cells (MDCK) to potassium oxalate (KOx), calcium oxalate monohydrate (COM) crystals, or a combination of the two was studied. The most noticeable effect of exposure of the cells to either KOx or COM crystals was loss of cells from the monolayer ranging from 20% to 30%, depending upon the particular treatment. Cellular enzyme values in the media were elevated significantly by 12 h of exposure, although in specific instances, elevated levels occurred at earlier time periods. As regards the monolayer, trypan blue exclusion was decreased significantly, although amounting to only a 4-5% reduction. Specific tritiated release occurred at 4 and 12 h after exposure to KOx and at 12 h after exposure to crystals. Structurally, COM-cell interactions were complex and extensive endocytosis was noted. Cells were released from culture either as cell-crystal complexes or from the intercellular spaces after exocytosis. When treatments were combined the effects were only slightly additive, but the two treatments potentiated each other: all media enzyme levels (with one exception) were elevated at 2 h, tritiated adenine release was present at 4 h, and there was more extensive cell loss from the culture monolayer. These data suggest that both KOx and COM crystals damage MDCK cells when applied alone, and in concert they act synergistically.

Adenine↗

A comparison of physical properties, oxalate-oxalic acid soluble substances, protein content, and in vitro protein digestibility of the blue-green alga Nostoc commune Vauch. from the Philippines and Japan.

The physical properties, amount of oxalate-oxalic acid soluble substances (OOSS), protein and in vitro protein digestibility of Nostoc commune from the Philippines and Japan were determined. The samples were the spherical type from the Philippines (PS), the dry field discoid type from the Philippines (PD), and the dry field discoid type from Japan (JD). The discoid types, PD and JD, regardless of origin showed higher settling volume in water (SV) and bulk density than PS. The OOSS values obtained for all samples were directly related to SV and bulk density. The WHC level was highest in PS. This desirable characteristic makes PS a technologically potential sample for large-scale food production applications in the future. Protein and in vitro protein digestibility varied significantly among all samples. In general, the algal samples examined possess desirable physical properties which are expected to be related to physiological functions.

Chemical Phenomena↗

Oxalate reduces calcium availability in the pads of the prickly pear cactus through formation of calcium oxalate crystals.

The pads (nopales) of the prickly pear cactus are considered to be a good source of minerals and other nutrients on the basis of compositional analysis. In this study, this analysis is taken a step further by assessing the availability of selected minerals in nopales using an in vitro digestion and dialysis method. The results obtained suggest that although nopales are enriched in a number of minerals, their tissue calcium is not freely available. Microscopic analysis, energy-dispersive X-ray microanalysis, and oxalate measurements suggest that this reduction in available calcium is a result of its sequestration in the form of calcium oxalate crystals. The issue of mineral availability in plant foods is important when the dependence of many populations around the world on plant foods as their main source of minerals and other nutrients is considered.

Biological Availability↗

Diphenyleneiodium (DPI) reduces oxalate ion- and calcium oxalate monohydrate and brushite crystal-induced upregulation of MCP-1 in NRK 52E cells.

BACKGROUND: Our earlier studies have demonstrated upregulation of monocyte chemoattractant protein-1 (MCP-1) in NRK52E rat renal epithelial cells by exposure to oxalate (Ox) ions and crystals of calcium oxalate monohydrate (COM) or the brushite (Br) form of calcium phosphate. The upregulation was mediated by reactive oxygen species (ROS). This study was performed to investigate whether NADPH oxidase is involved in ROS production. METHODS: Confluent cultures of NRK52E cells were exposed to Ox ions or COM and Br crystals. They were exposed for 1, 3, 6, 12, 24 and 48 h for isolation of MCP-1 mRNA and 24 h for enzyme-linked immunosorbent assay (ELISA) to determine the secretion of protein into the culture medium. We also investigated the effect of free radical scavenger, catalase, and the NADPH oxidase inhibitor diphenyleneiodium (DPI) chloride, on the Ox- and crystal-induced expression of MCP-1 mRNA and protein. The transcription of MCP-1 mRNA in the cells was determined using real-time polymerase chain reaction. Hydrogen peroxide and 8-isoprostane were measured to investigate the involvement of ROS. RESULTS: Exposure of NRK52E cells to Ox ions as well as the crystals resulted in increased expression of MCP-1 mRNA and production of the chemoattractant. Treatment with catalase reduced the Ox- and crystal-induced expression of both MCP-1 mRNA and protein. DPI reduced the crystal-induced gene expression and protein production but not Ox-induced gene expression and protein production. CONCLUSIONS: Exposure to Ox ions, and COM and Br crystals stimulates a ROS-mediated increase in MCP-1 mRNA expression and protein production. Reduction in ROS production, lipid peroxidation, low-density lipoprotein release, and inducible MCP-1 gene and protein in the presence of DPI indicates an involvement of NADPH oxidase in the production of ROS.

Animals↗

When aerosol sulfate goes up, so does oxalate: implication for the formation mechanisms of oxalate.

Oxalic acid is often the single most abundant water-soluble organic compound identified in ambient aerosols, but its precursors have not been identified, and its formation mechanism is not well understood. On the contrary, sulfate as a major aerosol component, its formation pathways have been established, and in-cloud processing is recognized as its major production pathway. Our measurements of aerosol sulfate and oxalate collected across a wide geographical span in the East Asia region, up to Beijing in the north and down to Hong Kong in the south, indicate that the two species are highly correlated among samples collected at the same location and among samples collected at different locations in the Pearl River Delta region. This good correlation is also found in measurements made elsewhere by other researchers. We argue that a common dominant formation pathway, likely in-cloud processing, explains the close tracking of the two chemically distinct species. This also highlights the potential importance of in-cloud processing as a pathway leading to formation of secondary organic aerosols.

Aerosols↗

Synovial fluid oxalate deposition complicating rheumatoid arthritis with amyloidosis and renal failure. Demonstration of intracellular oxalate crystals.

A patient with quiescent rheumatoid arthritis, amyloidosis and chronic renal failure developed an inflamed knee. Intracellular bipyramidal crystals characteristic of oxalate were found and are suggested as the cause of the acute arthritis. Since the patient had been treated with vitamin C, this precursor of oxalate is proposed as a possible factor in the crystal deposition.

Amyloidosis↗

[Calcium oxalate activity products and discriminant analysis of urinary parameters as basis for a calcium oxalate screening program (author's transl)].

The values of 7 urinary parameters in divided urine collections were measured in 20 patients with calcium oxalate stones and ten normal persons. On this basis the thermodynamic stone formation risk was calculated for the different urine collections. Stone formation risk in the 0-6 a.m. urine fractions appeared to be the most important. The risk of stone formation is significantly increased in the time from midnight to 6 a.m. To detect the risk situation the determination of oxalic acid, calcium, potassium and inorganic sulfate in night urine is adequate.

Calcium↗

A comparative study between etiological factors of calcium oxalate monohydrate and calcium oxalate dihydrate urolithiasis.

A comparative study between different etiological factors of calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD) is presented. The most frequent alteration in COD urolithiasis was associated with hypercalciuria, whereas in COM urolithiasis was associated with urinary pH. A comparison between COM and COD groups of stone formers that exhibited 1, 2 or 3 alterations was performed. Thus, in individuals with two simultaneous alterations, the association between altered urinary pH and hypomagnesiuria was the most frequent in the COM group, whereas the association between hypercalciuria and altered urinary pH was most frequent in the COD group. In individuals with three simultaneous alterations, the association between hypercalciuria, hyperphosphaturia and hyperuricuria was most frequent in both COM and COD stone formers.

Analysis of Variance↗

Molecule-based magnets formed by bimetallic three-dimensional oxalate networks and chiral tris(bipyridyl) complex cations. The series [ZII(bpy)3][ClO4][MIICrIII(ox)3] (ZII = Ru, Fe, Co, and Ni; MII = Mn, Fe, Co, Ni, Cu, and Zn; ox = oxalate dianion).

The synthesis, structure, and physical properties of the series of molecular magnets formulated as [ZII(bpy)3][ClO4][MIICrIII(ox)3] (ZII = Ru, Fe, Co, and Ni; MII = Mn, Fe, Co, Ni, Cu, and Zn; ox = oxalate dianion) are presented. All the compounds are isostructural to the [Ru(bpy)3][ClO4][MnCr(ox)3] member whose structure (cubic space group P4(1)32 with a = 15.506(2) A, Z = 4) consists of a three-dimensional bimetallic network formed by alternating MII and CrIII ions connected by oxalate anions. The identical chirality (lambda in the solved crystal) of all the metallic centers determines the 3D chiral structure adopted by these compounds. The anionic 3D sublattice leaves some holes where the chiral [Z(bpy)3]2+ and ClO4- counterions are located. These compounds behave as soft ferromagnets with ordering temperatures up to 6.6 K and coercive fields up to 8 mT.

Journal Article↗

Heptacoordinated Mn(II) in oxalate-based bimetallic 2D magnets: synthesis and characterisation of [Mn(L)6][Mn(CH3OH)M(III)(ox)3]2 (M(III) = Cr, Rh; ox = oxalate dianion; L = H2O, CH3OH).

Oxalate-based magnets have been known with several different crystallographic structures, from 1D to 3D, but with all of them based in metal ions with octahedral coordination. In this article we report a new bidimensional oxalate-bridged bimetallic magnet where the divalent metal appears heptacoordinated, which has strong effects in the structure and properties of this materials.

Journal Article↗

Alpha-1-microglobulin: inhibitory effect on calcium oxalate crystallization in vitro and decreased urinary concentration in calcium oxalate stone formers.

In the past few years, alpha-1-microglobulin (alpha1m) has been copurified from human urine with bikunin, a potent inhibitor of calcium oxalate (CaOx) crystallization in vitro. In this study, we have purified alpha1m without bikunin contamination and investigated its possible role in CaOx crystallization by in vitro and in vivo studies. Alpha-1m was purified with an anti-alpha1m antibodies CNBr-activated sepharose column. Two molecular species of alpha1m of respectively 30 and 60 kDa were purified. For each protein, two blots of 30 and 60 kDa cross-reacted with anti-alpha1m antibodies, suggesting that these two forms were derived one from the other. Both protein species inhibited CaOx crystallization in a dose-dependent manner in two in vitro tests. In the first test, the presence of alpha1m of 30 kDa (8 microg/ml) in a medium containing 0. 76 mM CaCl(2) (with (45)Ca) and 0.76 mM Ox(NH(4))(2) inhibited CaOx crystallization by 38% as estimated by supernatant radioactivity after 1 h of agitation. In the second test, CaOx kinetics were examined for 3 to 10 min in a turbidimetric model at 620 nm. The presence of alpha1m of 30 kDa in a medium containing 4 mM CaCl(2) and 0.5 mM Na(2)Ox inhibited CaOx crystallization by 41.5%, as estimated by the slope modification of turbidimetric curve. Alpha-1m can be considered as another inhibitor of urinary CaOx crystal formation, as shown by the present in vitro studies. Using an ELISA assay, we found that urinary alpha1m concentration was significantly lower in 31 CaOx stone formers than in 18 healthy subjects (2.95 +/- 0.29 vs 5.34 +/- 1.08 mg/l respectively, P = 0.01). The decreased concentration of alpha1m in CaOx stone formers could be responsible in these patients, at least in part, for an increased risk of CaOx crystalluria.

Adolescent↗

Development of a reference method for determining urinary oxalate by means of isotope dilution-mass spectrometry (ID-MS) and its usefulness in testing existing assays for urinary oxalate.

An accurate and reproducible GC-MS reference method for determining urinary oxalate is presented. With forty 24-h urines, comparisons were made between this reference method and four other existing procedures: a GC method, an HPLC method and two enzymatic assays. The results of the first two methods were in accordance with the GC-MS method. The performance of the enzymatic kits was less satisfactory. The use of a GC-MS reference method in evaluating existing and newly developed methods is recommended.

Chromatography, High Pressure Liquid↗