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To reduce the global CO 2 footprint of plastics, bio- and CO 2 -based feedstock are considered the most important design features for plastics. Oxalic acid from CO 2 and isosorbide from biomass are interesting rigid building blocks for high T g polyesters. The biodegradability of a family of novel fully renewable (bio- and CO 2 -based) poly(isosorbide- co -diol) oxalate (PISOX-diol) copolyesters was studied. We systematically investigated the effects of the composition on biodegradation at ambient temperature in soil for PISOX (co)polyesters. Results show that the lag phase of PISOX (co)polyester biodegradation varies from 0 to 7 weeks. All (co)polyesters undergo over 80% mineralization within 180 days (faster than the cellulose reference) except one composition with the cyclic codiol 1,4-cyclohexanedimethanol (CHDM). Their relatively fast degradability is independent of the type of noncyclic codiol and results from facile nonenzymatic hydrolysis of oxalate ester bonds (especially oxalate isosorbide bonds), which mostly hydrolyzed completely within 180 days. On the other hand, partially replacing oxalate with terephthalate units enhances the polymer's resistance to hydrolysis and its biodegradability in soil. Our study demonstrates the potential for tuning PISOX copolyester structures to design biodegradable plastics with improved thermal, mechanical, and barrier properties.

Oxalate is a divalent organic anion that affects many biological and commercial processes. It is derived from plant sources, such as spinach, rhubarb, tea, cacao, nuts, and beans, and therefore is commonly found in raw or processed food products. Oxalate can also be made endogenously by humans and other mammals as a byproduct of hepatic enzymatic reactions. It is theorized that plants use oxalate to store calcium and protect against herbivory. Clinically, oxalate is best known to be a major component of kidney stones, which commonly contain calcium oxalate crystals. Oxalate can induce an inflammatory response that decreases the immune system's ability to remove renal crystals. When formulated with platinum as oxaliplatin (an anticancer drug), oxalate has been proposed to cause neurotoxicity and nerve pain. There are many sectors of industry that are hampered by oxalate, and others that depend on it. For example, calcium oxalate is troublesome in the pulp industry and the alumina industry as it deposits on machinery. On the other hand, oxalate is a common active component of rust removal and cleaning products. Due to its ubiquity, there is interest in developing efficient methods to quantify oxalate. Over the past four decades, many diverse methods have been reported. These approaches include electrochemical detection, liquid chromatography or gas chromatography coupled with mass spectrometry, enzymatic degradation of oxalate with oxalate oxidase and detection of hydrogen peroxide produced, and indicator displacement-based methods employing fluorescent or UV light-absorbing compounds. Enhancements in sensitivity have been reported for both electrochemical and mass-spectrometry-based methods as recently as this year. Indicator-based methods have realized a surge in interest that continues to date. The diversity of these approaches, in terms of instrumentation, sample preparation, and sensitivity, has made it clear that no single method will work best for every purpose. This review describes the strengths and limitations of each method, and may serve as a reference for investigators to decide which approach is most suitable for their work.

The type strains of all 33 species in the genus Kribbella were tested for growth on oxalate ( - OOC-COO - ) as sole carbon source. Media were initially formulated to contain sodium oxalate, but even a concentration as low as 7.5 mM oxalate prevented growth. A modified medium based on calcium oxalate was very successful in characterising oxalate utilisation by Kribbella strains (metabolism of oxalate by oxalotrophic bacteria results in visible zones of clearing around the growth streaks on the opaque plates). To assess the variability of oxalate utilisation in Kribbella species, we also tested eight non-type strains for their ability to use oxalate. Thirty of 33 type strains (90.9%) and six of eight non-type strains (75%) were able to use oxalate as a sole carbon source. Based on these results, we propose that oxalate would be an excellent carbon source for the selective isolation of Kribbella strains. Based on the oxalate-utilisation phenotype and analyses of the 19 publicly available Kribbella type-strain genome sequences, we propose a pathway for oxalate metabolism in Kribbella. This pathway is significantly different from those previously proposed for oxalate metabolism in other bacteria, involving the indirect catabolism of oxalate to formate. Formate production is proposed to be involved in energy generation and to be crucial for oxalate import via an oxalate:formate antiporter. To our knowledge, this is the first report of an oxalate:formate antiporter in an aerobic, Gram-positive bacterium., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

The effect of ascorbic acid [AA (40 mmol L -1 )] and oxalic acid [OA (2 mmol L -1 )] on browning of litchi fruit was investigated under 5% CO 2  + 1% O 2 controlled atmosphere (CA) and compared with air at 5 ± 1 °C for 28 days. The combined application of AA and OA suppressed browning index, soluble quinones, and activities of polyphenol oxidase and peroxidase under CA compared with control. The combination of CA along with AA + OA reduced weight loss and maintained higher anthocyanins, total phenolics, membrane integrity, ascorbate peroxidase, catalase, glutathione reductase and superoxide dismutase activities compared with control. In addition, AA + OA + CA combination showed markedly lower malondialdehyde, superoxide anion and hydrogen peroxide with substantially higher soluble solids content, ascorbic acid, titratable acidity and sensory quality compared with control. In conclusion, AA + OA combination could be considered appropriate to delay browning and to conserve litchi fruit visual appearance under CA storage conditions., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Polycyclic aromatic hydrocarbons (PAHs) are on the list of priority pollutants to be eliminated from the environment due to their carcinogenic and mutagenic action, chemical stability, and resistance to biodegradation. The aim of this study was to evaluate the degradation of fluorene, a well-known PAH, in aqueous solutions (0.03 and 0.08 mg L -1 ), by means of a solar-driven conventional (PF) and modified photo-Fenton mediated by ferrioxalate complexes (PFF). Photolysis was also employed for comparison purposes. PF reaction was evaluated at different pH values (2.8, 3.5, and 4.0) and iron concentrations (2, 5, 10, and 20 mg L -1 ). On the other hand, PFF studies were conducted at mild pH conditions (4.0, 5.0, and 6.0) and iron content of 2 mg L -1 , keeping initial iron/oxalate molar ratio at 1:3. In both PF and PFF, the initial hydrogen peroxide/iron molar ratio was maintained at 5. In the presence of methanol as cosolvent for fluorene dissolution, the PF reaction was hampered and no consumption of H 2 O 2 was observed during the reaction carried out at constant pH (2.8). This led to low degradation rates, similar to those achieved by photolysis. Under the same pH but using acetonitrile as cosolvent for fluorene dissolution, fluorene degradation was found to be proportional to the iron content used in the PF experiments. On the other hand, at an invariable iron concentration of 5 mg Fe 2+ L -1 , the increase in pH was accompanied by a decrease in the molar fraction of the most photoactive iron complex (FeOH 2+ ) and ferric hydroxides precipitation, leading to a reduction in the fluorene degradation rate. With regard to the PFF tests, similar fluorene degradation performance was achieved at pH 4 and 5, while at pH 6 iron precipitation became relevant and the degradation rate was slightly slower. PFF has shown to be more efficient than the PF under the same pH (4) and iron concentration (2 mg L -1 ). Moreover, even at near neutral pH (6), fluorine degradation was shown to be feasible by using ferrioxalate complexes.

In this study, the photooxidation capacity of UVC/Oxalate (Ox) was evaluated using As(III) as a typical pollutant. The results show that the direct oxidation amount of As(III) induced by UVC in water was negligible, but the presence of Ox remarkably accelerated the oxidation rate of As(III). Under UVC irradiation, 50 μM As(III) can be completely oxidized to As(V) in the case of Ox concentration above 300 μM within 60 min. As(III) oxidation was found greatly related with the photodecomposition of Ox. Much more Ox can be mineralized in more acidic solution. At the same time, the photooxidation of As(III) was significantly favored at decreased initial pH from 8.0 to 3.0. In this reaction system, the role of oxygen was indispensable for Ox photodecomposition and As(III) photooxidation, which can be ascribed to its special roles as a precursor of reactive superoxide and an electron acceptor. In oxygen-present atmosphere, the in situ production of H 2 O 2 was detected during the photolysis of Ox and its photolysis product, i.e., OH primarily contributed to the oxidation of As(III). However, the photodecomposition of Ox and photooxidation of As(III) were significantly inhibited in the anaerobic environment. In general, the homogeneous photolysis of Ox in many commonly practiced UVC oxidation processes can be also proposed as a supplementary method of generating highly oxiditive species in aerobic condition., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Oxalic acid is widely distributed in biological systems and known to play functional roles in plants. The gene AAE3 was recently identified to encode an oxalyl-CoA synthetase (OCS) in Arabidopsis that catalyses the conversion of oxalate and CoA into oxalyl-CoA. It will be particularly important to characterise the homologous gene in rice since rice is not only a monocotyledonous model plant, but also a staple food crop. Various enzymatic and biological methods have been used to characterise the homologous gene. We first defined that AAE3 in the rice genome (OsAAE3) also encodes an OCS enzyme. Its K m for oxalate is 1.73 ± 0.12 mm, and V m is 6824.9 ± 410.29 U·min -1 ·mg protein -1 . Chemical modification and site-directed mutagenesis analyses identified thiols as the active site residues for rice OCS catalysis, suggesting that the enzyme might be regulated by redox state. Subcellular localisation assay showed that the enzyme is located in the cytosol and predominantly distributed in leaf epidermal cells. As expected, oxalate levels increased when OCS was suppressed in RNAi transgenic plants. More interestingly, OCS-suppressed plants were more susceptible to bacterial blight but more resistant to Al toxicity. The results demonstrate that the OsAAE3-encoded protein also acts as an OCS in rice, and may play different roles in coping with stresses. These molecular, enzymatic and functional data provide first-hand information to further clarify the function and mechanism of OCS in rice plants., (© 2017 The Authors. Plant Biology published by John Wiley & Sons Ltd on behalf of German Botanical Society, Royal Dutch Botanical Society.)

Oxalic acid, an effective metal-chelating ligand, is abundant in natural resources. In this study, a chitosan-oxalate complex biosorbent (COCB) was prepared by an iontropic cross-linking method. The COCB beads were characterized by Fourier transform infrared (FT-IR) spectra, X-ray diffraction (XRD) and thermogravimetric analysis (TGA). The XRD analysis shows that Cu(II) ions can form complexes with chitosan and oxalate. Adsorption of Cu(II) ions onto COCB beads was pH-dependent. The isothermal adsorption data fitted well to Langmuir equation with the maximum adsorption capacities of 227.27 mg/g for porous COCB beads and 175.44 mg/g for non-porous COCB beads at pH 5.0. The adsorption kinetics described by the pseudo-second-order diffusion models, suggesting that the rate-limiting step in adsorption was chemical sorption. Thermodynamic parameters (ΔG°<0 and ΔH°>0) indicated a spontaneous and endothermic adsorption process. The COCB bioadsorbent exhibited fast adsorption rate and high adsorption capacity for Cu(II) uptake., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The photocatalytic activity of a brick grain-supported iron oxide (denoted as B1) was tested for its activity to degrade Reactive Black 5 (RB5) in the presence of oxalic acid. B1 was obtained as a solid waste from a wastewater treatment plant, and characterized using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), X-ray powder diffraction (XRD) and N(2) adsorption/desorption isotherm analyses. The decolorization experiments were performed in a fluidized bed reactor with aeration under UV-A irradiation (λ = 365 nm). The effects of various factors such as solution pH, concentration of oxalic acid and dissolved oxygen on the decolorization of RB5 were evaluated considering the contributions of adsorption and photo-catalytic degradation. The role of dissolved iron in the removal of RB5 and the stability of B1 were also examined. In addition, the removal of TOC during the photo-catalytic reaction was monitored., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Teas prepared from the fruits of Ammi visnaga L. (syn. "Khella") have been traditionally used in Egypt as a remedy to treat kidney stones. It was the aim of our study to evaluate the effect of a Khella extract (KE) as well as the two major constituents khellin and visnagin on renal epithelial injury using LLC-PK1 and Madin-Darby-canine kidney (MDCK) cells. Both cell lines provide suitable model systems to study cellular processes that are possibly involved in the development of a renal stone. LLC-PK1 and MDCK cell lines were exposed to 300 microM oxalate (Ox) or 133 microg/cm(2) calcium oxalate monohydrate (COM) in presence or absence of 10, 50, 100 or 200 microg/mL KE. To evaluate cell damage, cell viability was assessed by determining the release of lactate dehydrogenase (LDH). KE (e.g. 100 microg/ml) significantly decreased LDH release from LLC-PK1 (Ox: 8.46+0.76%; Ox + 100 microg/ml KE: 5.41+0.94%, p<0.001) as well as MDCK cells (Ox: 30.9+6.58%; Ox+100 microg/ml KE: 17.5+2.50%, p<0.001), which indicated a prevention of cell damage. Similar effects for KE were observed in both cell lines when COM crystals were added. In LLC-PK1 cells khellin and visnagin both decreased the % LDH release significantly in cells that were pretreated with Ox or COM crystals. However, khellin and visnagin exhibited different responses in MDCK cells. Whereas khellin slightly reduced the % LDH release after exposure of the cells to Ox and COM crystals, visnagin significantly decreased % LDH release only after COM crystal exposure. Overall both compounds were more active in LLC-PK1 than in MDCK cells. In summary, exposure of renal epithelial cells to Ox or COM crystals was associated with a significant release of LDH indicating cell injury. Our data demonstrate that KE as well as khellin and visnagin could prevent renal epithelial cell damage caused by Ox and COM and could therefore play a potential role in the prevention of stone formation associated with hyperoxaluria., ((c) 2009 Elsevier GmbH. All rights reserved.)

The photolysis and photo-catalysis of ferrioxalate in the presence of hydrogen peroxide with UV irradiation (UV/ferrioxalate/H(2)O(2) process) for treating the commercial azo dye, reactive Black B (RBB), is examined. An effort is made to decolorize textile effluents at near neutral pH for suitable discharge of waste water. pH value, light source, type of initial catalyst (Fe(3+) or Fe(2+)) and concentration of oxalic acid (Ox) strongly affected the RBB removal efficiency. The degradation rate of RBB increased as pH or the wavelength of light declined. The optimal molar ratio of oxalic acid to Fe(III) is three, and complete color removal is achieved at pH 5 in 2h of the reaction. Applying oxalate in such a photo process increases both the RBB removal efficiency and the COD removal from 68% and 21% to 99.8% and 71%, respectively.

Purpose: Oxalate bioavailability is an important determinant of whether the consumption of a particular food is a high risk in individuals predisposed to kidney stones. We estimated and compared oxalate absorption from a high oxalate containing legume (black beans) and a high oxalate containing nut (almonds). We also compared an isotope method using extrinsically labeled oxalate and an oxalate load method to assess oxalate absorption., Materials and Methods: Six male and 5 female subjects participated in the 4 oxalate load tests, namely almonds, almonds with 20 mg C2-oxalic acid, black beans and black beans with 20 mg C2-oxalic acid. Each treatment provided a total of 120 mg oxalate, after which timed urine samples were collected for the analysis of oxalate, calcium and creatinine., Results: Average oxalate absorption from the 2 almond treatments (5.9%) using the oxalate load method was significantly higher than that from the 2 black bean treatments (1.8%) during the 24-hour post-oxalate load collection period. In contrast, C2-oxalic acid absorption from the almond (7.9%) and black bean (8.6%) treatments did not significantly differ., Conclusions: The higher oxalate absorption from almonds than from black beans suggests that the relative amount of soluble and insoluble oxalate in food has an important role in the determination of oxalate absorption. Since extrinsically provided C2-oxalate and oxalate naturally occurring in the high oxalate test foods appeared to be differentially absorbed, the data do not support the use of extrinsically labeled oxalate to assess food oxalate absorption.

Analysis of hydropathy suggests that in OxlT, the oxalate/formate antiporter of Oxalobacter formigenes, lysine 355 is within transmembrane helix no. 11. To test this idea, we used single-cysteine, histidine-tagged OxlT variants to study the organization of a 30-residue segment (residues 344-373) containing this region. Topology was examined by probing the A345C and A370C proteins with Oregon Green maleimide carboxylic acid, an impermeant and fluorescent thiol-reactive agent. Examination of purified protein showed that only A370C was fluorescent after treating intact cells with the probe, while both proteins were modified in tests with isolated membrane ghosts. In addition, labeling of A370C, but not A345C, was blocked when external cysteines were protected with the impermeant and nonfluorescent agent, methanethiosulfonate ethyltrimethylammonium. These findings confirm that A345 faces the cytoplasm, while A370C faces the periplasm. A similar study focused on 13 single-cysteine variants positioned throughout the target segment. That work revealed a striking discontinuity in reactivity toward Oregon Green maleimide; cysteines within a 10-residue central core (residues 351-360) were not labeled when membranes were probed, but were readily modified after protein denaturation. We suggest this core resides within the lipid bilayer, unavailable to an impermeant reporter. Since this region includes position 355, we also suggest that lysine 355 lies within the OxlT hydrophobic sector, where it may facilitate the binding and translocation of the anionic substrates, oxalate and formate.

Statement of Problem: New acids such as oxalic acid have been introduced as a conditioning agent in the total-etch technique. There is concern about long-term retention of the acid on enamel in relation to the superficial etch effect., Purpose: This in vivo study evaluated the marginal adaptation to enamel conditioned with either and oxalic acid solution or a phosphoric acid with SEM replica technique., Material and Methods: Twenty-four patients received one of each of three class III restorations. Two cavity preparations were pretreated with aluminum nitrate/oxalic acid/glycine solution 1 of the Gluma 2000 system. The first cavity was primed and sealed with Gluma 2000 solution 2, the second cavity with Gluma 3 and 4. The third cavity was conditioned with phosphoric acid (Gluma 1) and sealed with the bonding resin Gluma 4. All three cavities were restored with a hybrid resin composite (Pekafill). At baseline and after 1 year, replica impressions were made to study the margins with SEM. Semiquantitative analysis of the enamel interfaces was performed (x200 and x1000 magnifications). Marginal quality of the three restorative systems were compared and tested intraindividually., Results: The three restorations exhibited good enamel marginal adaptation and a high percentage of gap-free margins at baseline, 96% to 97% of the total length of margins investigated. Marginal quality decreased significantly after 1 year for all three groups. Gap-free margins were observed in 81% to 85% of the marginal length. No significant differences were found among the groups., Conclusion: Despite a less pronounced etch pattern created by conditioning of enamel with the oxalic acid solution, a good enamel marginal quality was observed at both evaluation times, comparable to the marginal adaptation of the phosphoric acid conditioned cavities.

The water proton relaxation rate measurements between 0.01 and 50 MHz on water solutions containing the cyanide adduct of the manganese-depleted manganese peroxidase (MnP-CN-) and increasing amounts of Mn2+ have been determined. The proton relaxivity curves have shown evidence of the formation of the protein/Mn2+ complex and have been analyzed in order to obtain spin Hamiltonian parameters and correlation times. Oxalate is shown not to alter the above profiles. This suggests that no protein-Mn2+-oxalate ternary complex is formed and that oxalate does not remove Mn2+ from the protein. On the basis of high-resolution 1H NMR experiments, we propose that Ce3+ and Gd3+ bind at the manganese site, and, on the basis of the charge, we propose that they may mimic Mn3+. The water proton relaxation rates of water solutions containing manganese-depleted MnP-CN- and increasing amounts of Gd3+ have been measured and analyzed. Oxalate is shown to remove the trivalent metal ions. This suggests that trivalent metal ions bind oxalate and diffuse away from the protein presumably as oxalate complexes. Implications for the enzymatic mechanism are discussed.

Pyruvate kinase from rabbit muscle has been cocrystallized as a complex with MgIIATP, oxalate, Mg2+, and either K+ or Na+. Crystals with either Na+ or K+ belong to the space group P2(1)2(1)2(1), and the asymmetric units contain two tetramers. The structures were solved by molecular replacement and refined to 2.1 (K+) and 2.35 A (Na+) resolution. The structures of the Na+ and K+ complexes are virtually isomorphous. Each of the eight subunits within the asymmetric unit contains MgIIoxalate as a bidentate complex linked to the protein through coordination of Mg2+ to the carboxylates of Glu 271 and Asp 295. Six of the subunits also contain an alpha,beta,gamma-tridentate complex of MgIIATP, and the active-site cleft, located between domains A and B, is closed in these subunits. In the remaining two subunits MgIIATP is missing, and the active-site cleft is open. Closure of the active-site cleft in the fully liganded subunits includes a rotation of 41 degrees of the B domain relative to the A domain. alpha-Carbons of residues in the B domain undergo movements of up to 17.8 A (Lys 124) in the cleft closure. Lys 206, Arg 119, and Asp 177 from the B domain move several angstroms from their positions in the open conformation to contact the MgIIATP complex in the active site. The gamma-phosphate of ATP coordinates to both magnesium ions and to the monovalent cation, K+ or Na+. A Mg2+-coordinated oxygen from the MgIIoxalate complex lies 3.0 A from Pgamma of ATP, and this oxygen is positioned for an in-line attack on the phosphorus. The side chains of Lys 269 and Arg 119 are positioned to provide leaving-group activation in the forward and reverse directions. There is no obvious candidate for the acid/base catalyst near the 2-si face of the prospective enolate of the normal substrate. A functional group linked through solvent and side-chain hydroxyls may function in a proton relay.

Oxalate transport across the contraluminal membrane of the proximal tubule was studied in vivo using the "capillary stopped flow microperfusion method" (Pflügers Arch 400:250-256, 1984). Cellular uptake of oxalate was characteristic of a carrier-mediated transport process (Jmax = 1.6 +/- 0.6 pmol/s per cm proximal tubular length, Km = 2.03 +/- 0.77 mmol/l). Sulphate inhibited oxalate transport in a dose-dependent manner (Ki-value = 1.53 +/- 0.38 mmol/l). Sulphate transport across the basolateral membrane was also characteristic of a carrier- mediated transport process (Jmax = 1.83 +/- 0.56 pmol/s per cm proximal tubular length, Km = 1.37 +/- 0.57 mmol/l). Oxalate inhibited the sulphate transport in a dose-dependent manner (Ki = 2. 06 +/- 0.82 mmol/l). No significant differences were found between the Ki values and the Km values of the two substances, indicating that oxalate and sulphate are transported by the same carrier across the basolateral membrane of the proximal tubule. Oxalate transport was not dependent on the extracellular sodium or potassium concentration. Bicarbonate competitively inhibited the oxalate transport. Chloride significantly inhibited the oxalate transport, but not dose dependently. It is, therefore, suggested that oxalate is transported into the cell of the proximal tubule in exchange for sulphate or bicarbonate. The dose-independent inhibition by chloride is suggested to be mediated by the coupling of the sulphate (bicarbonate)/oxalate exchanger with the chloride/bicarbonate exchanger at the basolateral membrane of the proximal tubule. This, furthermore, suggests that the transport of oxalate or sulphate across the basolateral membrane might be indirectly coupled with the reabsorption of chloride at this membrane side.

To improve our understanding of both diagnosis and treatment of diseases of oxalate metabolism, we first set out to establish a new ion-chromatographic method to determine normal plasma levels of oxalate, citrate and sulfate from single plasma samples. In 50 infants and children (23 girls, 27 boys, aged 0.2 to 17 years) with normal renal function, blood was drawn in Li-heparin tubes, placed on ice and preserved immediately with 40 microliters M HCl/ml plasma in two ultracentrifugation steps. For measurement, plasma was injected onto an ion chromatography system with NaOH as the mobile phase, and then run as a linear gradient from 5 mM to 52.5 mM over 21 minutes. Analysis yielded measurable and reproducible oxalate (6.43 +/- 1.06 microM/liter), citrate (79.3 +/- 27.4 microM/liter) and sulfate (235.0 +/- 85.3 microM/liter) levels, without any age and gender specific differences. The least detectable plasma oxalate level was < 0.3 microM with a high reliability and reproducibility (coefficient of variance 1.95 to 4.75%). In conclusion, we established a reproducible, precise method to determine the relevant plasma anions involved in mineral metabolism, which heretofore have not been easily measurable. Studies of diseases of oxalate and citrate metabolism are ongoing on the basis of the normal plasma values achieved in this study.

Within the overall objective of whether ruminants are able to form conditioned aversions (CFA) toward a food flavour associated with the administration of an aversive stimulus which occurs naturally in food plants (oxalic acid, OA), two specific objectives were tested: (1) whether the rate and degree of formation of CFA are dependent on the dose rate of OA administered and (2) whether the persistence of formed CFA depends on the previous dose rate of OA. Sheep were conditioned to associate the specific flavour of one of two novel foods with either the oral administration of OA or equivalent placebos. Four dose rates of OA were tested (0.06, 0.12, 0.18 and 0.24 g/kg sheep live weight per d), with twelve sheep per dose. Each conditioning period lasted for 8 d and was repeated four times. At the end of each conditioning period the preference for the two flavours was measured in short-term, 20 min preference tests. The persistence of the CFA was measured at 0, 7, 21 and 49 d after the completion of the conditioning phase with long-term, 3 h preference tests. The results of the experiment indicated that: (1) the rate and degree of formation of CFA were dependent on the rate of administration of OA; (2) sheep required repeated exposures to the lower dose rates of OA in order to develop CFA and these CFA did not persist in the absence of continual reinforcement; (3) CFA to the higher dose rates of OA were developed after as little as one exposure and persisted over a period of at least 7 weeks. These findings are consistent with the expectation that ruminants should be able to select a diet which minimizes the risk of consumption of potentially harmful foods, whilst at the same time maintaining a degree of flexibility in their feeding behaviour.

Tracer experiments in rats mimicking type II primary hyperoxaluria, with an expanded intracellular pool of hydroxypyruvate, showed that the excess formation of oxalate did not originate from its immediate precursor glyoxylate. In these animals, the hepatic and kidney activities of oxalate synthesising enzymes such as lactate dehydrogenase and glycolate oxidase were normal, but tissue lipid peroxidation was significantly higher. In vitro experiments established that in a mild alkaline solution, hydroxypyruvate underwent auto-oxidation to form oxalate and H2O2 and also inhibited lactate dehydrogenase and glycolate oxidase from oxidising glyoxylate to oxalate. On the basis of the experimental evidence, we suggest that in type II primary hyperoxaluria, the accumulating hydroxypyruvate could reduce the intracellular pool of glyoxylate and on ageing, give rise to excess oxalate and H2O2, to cause oxalosis in the former and free radical mediated-cell injuries in the latter.

The reaction of Fe(II) oxalate with hydrogen peroxide and dioxygen was studied for oxalate concentrations up to 20 mM and pH 2-5, under which conditions mono- and bis-oxalate complexes (Fe[II](ox) and Fe[II](ox)2[2-]) and uncomplexed Fe2+ must be considered. The reaction of Fe(II) oxalate with hydrogen peroxide (Fe2+ + H2O2 --> Fe3+ + .OH + OH-) was monitored in continuous flow by ESR with t-butanol as a radical trap. The reaction is much faster than for uncomplexed Fe2+ and a rate constant, k = 1 x 10(4) M(-1) s(-1) is deduced for Fe(II)(ox). The reaction of Fe(II) oxalate with dioxygen is strongly pH dependent in a manner which indicates that the reactive species is Fe(II)(ox)2(2-), for which an apparent second order rate constant, k = 3.6 M(-1) s(-1), is deduced. Taken together, these results provide a mechanism for hydroxyl radical production in aqueous systems containing Fe(II) complexed by oxalate. Further ESR studies with DMPO as spin trap reveal that reaction of Fe(II) oxalate with hydrogen peroxide can also lead to formation of the carboxylate radical anion (CO2-), an assignment confirmed by photolysis of Fe(II) oxalate in the presence of DMPO.

Crystals of the oxalic acid complex of DL-lysine (triclinic P1; a = 5.540(1), b = 10.764(2), c = 12.056(2) A, alpha = 77.8(1), beta = 80.6(1), gamma = 75.6(1).; R = 4.7% for 2023 observed reflections) contain lysine and semioxalate ions in the 1:1 ratio, whereas the ratio of lysine and semioxalate/oxalate ions is 2:3 in the crystals of the L-lysine complex (monoclinic P2(1); alpha = 4.906(1), b = 20.145(4), c = 12.455(1) A, beta = 92.5(1).; R = 4.4% for 1494 observed reflections). The amino acid molecule in the L-lysine complex has an unusual ionisation state with positively charged alpha- and side-chain amino groups and a neutral carboxyl group. The unlike molecules aggregate into separate alternating layers in the DL-lysine complex in a manner similar to that observed in several of the amino acid complexes. The L-lysine complex exhibits a new aggregation pattern which cannot be easily explained in terms of planar features, thus emphasizing the fundamental dependence of aggregation on molecular characteristics. Despite the differences in stoichiometry, ionisation state and long-range aggregation patterns, the basic element of aggregation in the two complexes exhibits considerable similarity.

We have studied the mechanisms of NaCl transport in the mammalian proximal tubule. We identified Cl(-)-formate and Cl(-)-oxalate exchangers as possible mechanism's of uphill Cl- entry across the apical membrane of proximal tubule cells. For steady state Cl- absorption to occur by these mechanisms, formate and oxalate must recycle from lumen to cell. Recycling of formate from lumen to cell may occur by H(+)-coupled formate transport and nonionic diffusion of formic acid in parallel with Na(+)-H+ exchange. Oxalate recycling from lumen to cell may take place by oxalate-sulfate exchange in parallel with Na(+)-sulfate cotransport. Cl- exit across the basolateral membrane is most likely mediated by Cl- channels. To identify the Na(+)-H+ exchanger (NHE) isoform(s) expressed on the brush border membrane of proximal tubule cells, we developed isoform-specific monoclonal and polyclonal antibodies. We found that NHE1 is present on the basolateral membrane of all nephron segments, whereas NHE3 is present on the apical membrane of cells in the proximal tubule and the thin and thick limbs of the loop of Henle. NHE3 is also present in a population of subapical intracellular vesicles, suggesting possible regulation by membrane trafficking. The inhibitor sensitivity of Na(+)-H+ exchange in renal brush border vesicles indicates that it is mediated by NHE3 under baseline conditions and during up-regulation by metabolic acidosis. Increased apical membrane Na(+)-H+ exchange activity in response to metabolic acidosis and during renal maturation is associated with increased NHE3 protein expression. Finally, we found that the organic anion-dependent absorption of Cl- is markedly down-regulated in metabolic acidosis in parallel with the up-regulation of brush border membrane Na(+)-H+ exchange. Thus, differential regulation of apical membrane ion exchangers may provide a mechanism to regulate the relative rates of NaHCO3 and NaCl reabsorption.

In this paper we present an improved method for the selective and sensitive determination of oxalate in different matrices such as urine, plasma, and food. The method uses ion chromatography for the separation of anions. To overcome problems with interfering matrix-anions, colourings, and macromolecules, we used an inline enzyme-reactor (ER) containing immobilised oxalate oxidase, which converts oxalate to hydrogen peroxide. Hydrogen peroxide was analysed with high sensitivity by amperometric detection. The determination limit for the HPLC-ER method was 1.5 mumol/1, the mean recovery in urine was 102%. The evaluation in a urinary matrix achieved C.V. values from 2.2% to 6.7% for the within-run precision and C.V. values from 3.7% to 8.6% for the between-batch precision. The results of the new method were statistically equivalent to those obtained by enzymatic kits. We present first results of the HPLC-ER method, when applied to body fluids and food analysis.

An experimental model that allowed differentiation between endogenously and exogenously derived urinary oxalate was used to assess the effect of different forms and doses of ingested calcium on oxalate absorption and excretion. In replication 1 (R-1), subjects participated in three oxalate load (OL) tests: baseline (OL alone), calcium carbonate (OL with concomitant calcium carbonate ingestion), and calcium citrate malate (CCM) (OL with concomitant CCM ingestion). The calcium salts each provided 300 mg elemental Ca. OLs consisted of 180 mg unlabeled and 18 mg 1,2[13C2]oxalic acid. In R-2, subjects participated in four OL tests: baseline (OL alone) and OLs administered concomitantly with 100, 200, or 300 mg Ca. Timed urine samples after the OL were collected at 2-h intervals for the initial 6 h and samples were pooled into 9-h aliquots for the remaining 18 h of the 24-h period. In R-1, 24-h mean exogenous oxalate decreased (P < 0.05) after the OL from 36.2 mg (baseline) to 16.1 mg (after calcium carbonate) and to 14.3 mg (after CCM) whereas endogenous oxalate remained relatively constant. Mean 24-h oxalate absorption decreased significantly from that at the time of the baseline treatment (18.3%) after both calcium carbonate (8.1%) and CCM (7.2%) treatments. In R-2, mean 24-h oxalate absorption was significantly lower after 200 (5.9%) and 300 (7.6%) mg Ca than after 100 mg Ca (9.1%) and the OL alone (11.3%). Concomitant meal ingestion significantly decreased oxalate absorption in the absence of dietary calcium but not in association with the 300-mg Ca treatment. The overall data provide definitive evidence that dietary calcium can reduce oxalate absorption and excretion. Calcium carbonate and CCM were equally effective in this regard and a minimum of 200 mg elemental Ca maximized this effect in conjunction with an oxalic acid intake of 198 mg.

Nutritional (ascorbic acid, dehydroascorbic acid and carotenes); antinutritional and toxic components (oxalic acid, nitrate and erucic acid) were determined in sixteen popular species of wild edible plants which are collected for human consumption in southeast Spain. Ascorbic + dehydroascorbic acids contents were very high in several species, especially in Chenopodium album L. (155 mg/100 g). Carotenoid content ranged from 4.2 mg/100 g (Stellaria media Villars) to 15.4 mg/100 g (Amaranthus viridis L.). A range of values was found for oxalic acid from absence to 1100 mg/100 g of plant material. Nitrate contents ranged from 47 mg/100 g (Salicornia europaea L.) to 597 mg/100 g (Amaranthus viridis L.). Low amounts of erucic acid were found in the Cruciferae family (Sisymbrium irio L. 1.73%; Cardaria draba L. 1.23%) and Plantago major L. 3.45%.

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.

Crystals of the oxalic acid complex of L-histidine (orthorhombic P2(1)2(1)2(1); a = 5.535(4), b = 6.809(4), c = 26.878(3) A; R = 3.6% for 1188 observed reflections) contain histidine molecules and semi-oxalate ions in the 1:1 ratio, while the ratio is 1:2 in the crystals of the DL-histidine complex (monoclinic P2(1)lc; a = 6.750(7), b = 10.139(2), c = 19.352(2) A, beta = 90.8 degrees; R = 3.7% for 3176 observed reflections). The histidine molecule in the latter has an unusual ionization state with positively charged amino and imidazole groups and a neutral carboxyl group. The molecule has the sterically least favourable allowed conformation with the side chain imidazole ring staggered between the alpha-amino and the alpha- carboxyl (carboxylate) groups, in both the structures. The unlike molecules aggregate into separate alternating layers in both of them. There are elements of similarity in the aggregation patterns in the semi-oxalate layers in the two complexes, but the patterns in the amino acid layers are entirely different. Interestingly, the crystal structure of L-histidine semi-oxalate has broad similarities with that of DL-histidine = glycolate, demonstrating how broad features of aggregation could be retained inspite of changes in chirality and composition. The unusual ionization state of the amino acid molecule in the DL-histidine complex is reflected in a hitherto unobserved aggregation pattern in its crystal structure.

17O NMR is complementary to 2H NMR for studies of hydrate structure and dynamics in solids, insofar as the quadrupole coupling parameters can be accurately determined. At a Larmor frequency of 34.2 MHz, the effects of a large 17O quadrupole coupling (e2qQ/h approximately 7 MHz) on both the orientational dependence of the central transition resonance frequency and its observation by cross-polarization are examined in powders and a single crystal of oxalic acid dihydrate. Comparison with the exact energies shows that the variation of the central transition frequency with tensor orientation is accurately described as a second-order perturbation, even when e2qQ/h and the Larmor frequency are comparable. The combined chemical and quadrupole-induced shifts of the central transition frequency are separated by their distinct transformation properties, and we report the H2(17)O quadrupole coupling tensor in alpha-oxalic acid dihydrate, which is compared with the hydrogen bonding structure. Excitation of the central transition by cross-polarization is examined in terms of the 17O nutation frequency. Distortions in powder patterns and the loss of lines in single-crystal spectra owing to the orientational dependence of the first-order quadrupole coupling are verified, and simple methods for mitigating the problem in single-crystal and powder experiments are presented.

Urinary oxalic acid excretion was examined in 61 patients with chronic nephropathies and in 21 patients after renal transplantation with a varying mean glomerular filtration. In both groups of patients a correlation was found between the plasma oxalic acid and serum creatinine and by a hyperbolic correlation between plasma oxalic acid and creatinine clearance. Moreover the authors found a direct correlation between oxalic acid and FEoxalic acid and FE(Na)+, FEH20 and FE1-ascorbic acid in both patient groups. Various chronic nephropathies and treatment in both groups did not affect the revealed correlations. In 13 healthy subjects during the period of maximum water diuresis urinary Na+ excretion did not increase but there was a significant increase of the l-ascorbic acid and oxalic acid excretion. In a group of 8 patients in the polyuric stage of chronic renal failure without dialysis treatment under conditions of increased dietary NaCl intake (15 g/24 h) a significant increase of the urinary Na+ excretion was recorded while the l-ascorbic acid and oxalic acid excretion was unaltered. From the assembled values of the examined biochemical indicators ensues that urinary oxalic acid excretion, similarly as ascorbic acid excretion, depended on water excretion.

Purpose: The aim of this investigation was to differentiate between aggregation and crystal growth by studying the structure of oxalate stones at high spatial resolution using recently developed microscopy techniques., Materials and Methods: Sections from 6 complete human oxalate stones and 4 stone fragments were prepared by ultramicrotomy and examined by both low voltage scanning electron microscopy and atomic force microscopy., Results: The scanning electron microscopy showed lamellar structures up to 10 microns. in size, consistent with previous results, and provided evidence that these structures were composed of smaller particles. The atomic force microscopy clearly showed arrays of the small particles, whose size varied between 500A and 2800A., Conclusion: Our images suggest that an ordered aggregation of small crystallites is responsible for oxalate stone formation.

Recent studies on LLC-PK1 cells demonstrated that oxalate, a simple dicarboxylic acid, acts as a mitogen for these renal epithelial cells. Exposure to oxalate initiates DNA synthesis, induces the expression of one of the early growth response genes c-myc and stimulates proliferation of quiescent cultures of LLC-PK1 cells. The present studies examined the possibility that expression of the c-myc protooncogene is obligatory for this mitogenic response. Specifically we determined whether pretreatment with c-myc antisense oligonucleotides would block the proliferative effects of oxalate in LLC-PK1 cells. Quiescent cultures of LLC-PK1 cells were exposed to oxalate in the presence and absence of c-myc antisense and the effects of oxalate on c-myc protein expression (Myc), DNA synthesis and cell growth were assessed. Exposure of cells to oxalate alone increased the expression of Myc within two hours. Pretreatment with c-myc antisense abolished this response. Further, pretreatment of cells with c-myc antisense but not nonsense oligonucleotides blocked the oxalate-induced initiation of DNA synthesis. Increases in cell number in response to oxalate (measured after 72 hr exposure) were also blocked by exposure to c-myc antisense. These findings suggest that c-myc gene expression is critical for the mitogenic effects of oxalate in LLC-PK1 cells.

Objective: Five methods for oxalate analysis in urine are compared with respect to reliability, accuracy, and practicability., Results: Suppressed and unsuppressed ionchromatography, as well as the enzymatic Sigma-Kit, achieve low coefficients of variation for the within-batch imprecision (1.1-8.0%) and between-day imprecision (1.6-7.2%). The results of these methods are comparable and the mean recovery rate ranges between 99.7% and 100.9%. The enzymatic Boehringer-Kit gives higher CV (3.1-9.5%) and the results are lower than those obtained by the methods mentioned above; the recovery rate is sufficient (92.4%)., Conclusion: The handling of the chromatographic methods is very easy, whereas the enzymatic methods require more manual work. In relation to sample throughput, charges for the enzymatic methods are about twice as high as for the chromatographic methods. In respect to reliability and accuracy, the chromotropic acid method cannot be recommended (recovery rate 68%).

The generation of transmembrane ion gradients by Oxalobacter formigenes cells metabolizing oxalate was studied. The magnitudes of both the transmembrane electrical potential (delta psi) and the pH gradient (internal alkaline) decreased with increasing external pH; quantitatively, the delta psi was the most important component of the proton motive force. As the extracellular pH of metabolizing cells was increased, intracellular pH increased and remained alkaline relative to the external pH, indicating that O. formigenes possesses a limited capacity to regulate internal pH. The generation of a delta psi by concentrated suspensions of O. formigenes cells was inhibited by the K+ ionophore valinomycin and the protonophore carbonyl cyanide-m-chlorophenylhydrazone, but not by the Na+ ionophore monensin. The H+ ATPase inhibitor N,N'-dicyclohexyl-carbodiimide inhibited oxalate catabolism but did not dissipate the delta psi. The results support the concept that energy from oxalate metabolism by O. formigenes is conserved not as a sodium ion gradient but rather, at least partially, as a transmembrane hydrogen ion gradient produced during the electrogenic exchange of substrate (oxalate) and product (formate) and from internal proton consumption during oxalate decarboxylation.

Background: Oxalic acid-degrading bacteria have been isolated from human faces. Therefore, the possibility exists that oxalic acid in food is degraded in human intestine by such bacteria, and absorption and excretion of oxalic acid is reduced. It may be possible that patients who form idiopathic stones have fewer oxalic acid-degrading bacteria than healthy controls. The purpose of this study is to examine the possibility that oxalic acid in food is degraded in the human intestine., Methods: Nineteen patients with calcium stones and 13 healthy subjects were included in the study. Samples of feces were diluted with Barber medium containing 1 g/L of oxalic acid dihydrate, and 1%, 0.1% and 0.01% suspensions were prepared. These solutions were incubated under anaerobic conditions at 37 degrees C for seven days. The degradation of oxalic acid was estimated by the decrease of oxalic acid in the medium., Results: The feces of almost all persons with or without calculi degraded oxalic acid to some degree. Incomplete or no oxalic acid degradation was found in 15 of the 19 stone-forming patients and in five of the 13 stone-free controls., Conclusion: Large numbers of oxalic acid-degrading bacteria were observed less often in the feces of stone-formers than in the feces of stone-free controls.

Primary hyperoxaluria is a rare hereditary disease. Two types have been identified. Type 1 is due to the deficiency of the liver-specific peroxisomal enzyme alanine:glyoxylate aminotransferase/serine: pyruvate amino-transferase whereas, in type 2, the deficiency concerns the glyoxylate reductase/D-glycerate dehydrogenase, a cytosolic enzyme present in the leucocytes and hepatocytes. In the elapsed decade, important progress in molecular biology led to the introduction of new strategies in the diagnosis and treatment of type 1 primary hyperoxaluria. However, the greater rarity of type 2 has so far prevented similar development. The present review recalls the normal metabolism of oxalic acid, details its deviations and their clinical consequences, and describes the methods of diagnosis and treatment to be presently recommended in primary hyperoxaluria.

Purpose: The present studies assessed the possibility that high concentrations of oxalate may be toxic to renal epithelial cells., Materials and Methods: Subconfluent cultures of LLC-PK1 cells were exposed to oxalate, and the effects on cell morphology, membrane permeability to vital dyes, DNA integrity and cell density were assessed., Results: Oxalate exposure produced time- and concentration-dependent changes in the light microscopic appearance of LLC-PK1 cells with higher concentrations ( > 140 microM.) inducing marked cytosolic vacuolization and nuclear pyknosis. Exposure to oxalate also increased membrane permeability to vital dyes, promoted DNA fragmentation and, at high concentrations (350 microM. free oxalate), induced a net loss of LLC-PK1 cells., Conclusions: Since high concentrations of oxalate can be toxic to renal epithelial cells, hyperoxaluria may contribute to several forms of renal disease including both calcium stone disease and end-stage renal disease.

45Ca2+ uptake and efflux studies were performed on membranes prepared from dog mesenteric artery and rat vas deferens. Oxalate-stimulated, ATP-dependent Ca2+ uptake in microsomal vesicles, a property characteristic of sarcoplasmic reticulum, was completely inhibited in a concentration-dependent manner by cyclopiazonic acid (0.1-30 microM) and thapsigargin (10 nM-10 microM). Using discontinuous sucrose gradient centrifugation, rat vas deferens microsomes were separated into two fractions, one enriched in plasma membrane (F2), the other enriched in sarcoplasmic reticulum (F3). The F3 fraction had a major increase in Ca2+ uptake in the presence of oxalate, which was completely inhibited by either cyclopiazonic acid or thapsigargin. In the F2 fraction Ca2+ uptake in the presence of oxalate was lower than in F3 and was not completely inhibited by thapsigargin and cyclopiazonic acid. Instead, the F2 fraction had a thapsigargin-insensitive and cyclopiazonic acid insensitive, saponin-sensitive component of uptake, which probably represents Ca2+ uptake by plasma membrane. In the absence of oxalate, the inhibition of Ca2+ uptake by saponin and cyclopiazonic acid or thapsigargin was additive in the F2 and F3 fractions, suggesting that cyclopiazonic acid and thapsigargin selectively inhibited sarcoplasmic reticulum derived Ca2+ uptake and did not affect plasma membrane derived Ca2+ uptake. Measurement of the initial rate of Ca2+ uptake in the presence and absence of oxalate by rat vas deferens microsomes demonstrated selective inhibition of oxalate-stimulated Ca2+ uptake by cyclopiazonic acid and thapsigargin. Ca2+ efflux from rat vas deferens microsomes actively loaded with 45Ca2+ either in presence or the absence of oxalate was not increased by cyclopiazonic acid or thapsigargin, showing that the inhibition of Ca2+ accumulation was not due to an increase in Ca2+ efflux. In both rat vas deferens and dog mesenteric artery, the maximal inhibitory effects of cyclopiazonic acid developed rapidly, whereas for maximal inhibition thapsigargin required pretreatment of microsomes prior to measurement of Ca2+ uptake. In rat vas deferens microsomes the inhibitory effects of cyclopiazonic acid could be quickly and completely reversed, whereas the effects of thapsigargin were not easily reversed. Collectively, these results suggest selectivity of cyclopiazonic acid and thapsigargin for the sarcoplasmic reticulum Ca2+ pump. Their selective inhibitory properties and differences in onset and offset of inhibition make cyclopiazonic acid thapsigargin useful pharmacological tools in the study of the physiological and pathophysiological roles of the sarcoplasmic reticulum Ca2+ pump in regulating smooth muscle Ca2+.

A simple method for the simultaneous determination of oxalic and citric acids had been developed using reversed-phase HPLC. An aqueous solution containing potassium dihydrogen phosphate (0.25%) and tetrabutylammonium hydrogen sulphate (2.5 mmol) at pH 2.0 was used as mobile phase. Under these conditions both the components were well resolved and detected at 210 nm. The recovery for oxalic and citric acids was 97% and 102%, respectively. The method presented here was applied to urine specimens of a large number of urolithic patients and control subjects. Because of the simplicity of the method its application provides better means of monitoring the concentration of oxalic and citric acids in the formation of renal calculi.

Spinach was evaluated for its bioavailability of magnesium in the experiment with magnesium-deficient rats. The effect of oxalic acid on absorption of dietary magnesium was also examined in the same experiment. After there were significant differences in the body weight of the rats between the control group and the magnesium-deficient group, and after the number of dead rats increased, the magnesium-deficient rats were divided into six groups. They were pair-fed for 8 days on the magnesium-deficient diet, magnesium-deficient diet supplemented with raw powdered spinach (R-sp), boiled powdered spinach (B-sp), or fried powdered spinach (F-sp), control diet supplemented with oxalic acid (Ox-C), and control diet (+Mg). On the 10th day, there was no significant difference in the food intake of the rats between the control group and magnesium-deficient group. However, the body weight, and body weight gain of the rats increased more significantly in the control group than in those of the magnesium-deficient group. Also, the contents of calcium and phosphorus in the liver and kidneys, and serum calcium content increased significantly in the magnesium-deficient rats compared with those of the control rats. However, the serum magnesium content decreased significantly in the magnesium-deficient rats. An especially large amount of calcium was accumulated in the kidneys of the magnesium-deficient rats. At the end of the experimental period, there were no significant differences in the food intake, body weight and body weight gain of the rats among the control group and each of the spinach-added groups. The body weight and body weight gain of the Ox-C rats decreased significantly in comparison with those of the control group and each of the spinach-added groups. Although, there were no significant differences in the concentrations of serum minerals (Mg, Ca and P) among each of the groups, kidney magnesium, calcium and phosphorus, and liver magnesium and phosphorus were significantly higher in each of the spinach added groups than those of the control, Ox-C and +Mg groups. A large amount of calcium was accumulated in the kidneys of the rats fed on the R-sp, B-sp, F-sp and Ox-C diets. However, the kidney calcium of each of the spinach-added groups markedly decreased in comparison with kidney calcium of the magnesium-deficient rats on the 10th day, when the magnesium-deficient rats were separated. There was no significant difference in the magnesium content of the left tibiae among each of the spinach-added groups. Also, the magnesium contents of the left tibiae of each of the additional groups did not reach the level of those of the control rats. The contents of calcium and phosphorus of the left tibiae were not significantly different among any of the groups except for both the R-sp and Ox-C groups, and decreased significantly in the R-sp and Ox-C groups compared with those of the other groups. A highly positive correlation between bone calcium and bone strength was not observed in this study; the breaking force of the left femurs of the B-sp and F-sp rats increased significantly in comparison with that of the Ox-C group. The rate of magnesium absorbed by the rats receiving the control, R-sp, B-sp, F-sp, Ox-C, and +Mg diets was 88.9, 80.2, 88.4, 90.4, 88.1, and 87.7%, respectively. The rate of apparent absorption of calcium from the control, Mg-deficient, R-sp, B-sp, F-sp, Ox-C and +Mg diet was 87.0, 84.1, 57.3, 66.4, 66.2, 53.3 and 83.5%, respectively. The data indicate that oxalic acid remained in spinach after cooking of boil or frizzle was not deleterious to magnesium availability, and that spinach is one of the most promising sources of magnesium.

Wild-type Aspergillus niger N402 and glucoamylase++ overproducing transformant A. niger N402[pAB6-10]B1 have grown in maltodextrin- and xylose-limited recycling culture at pH 4.5 on mineral medium. The only products formed were organic acids and proteins, among which glucoamylase. The production of organic acids by the fungus has been analyzed qualitatively and quantitatively using capillary electrophoresis. The only organic acids produced in these cultures were substantial amounts of citric acid. This is the first demonstration of abundant oxalic acid production and a very low citric acid production by submerged cultures of A. niger. In the maltodextrin-limited culture the oxalic acid production rate increased during the first 80 h of cultivation and decreased after that time. In xylose-limited recycling culture the oxalic acid production rate always increased in time and highest values were found in the last samples taken from the culture after about 140 h of cultivation. Oxalic acid production rates were highest by the wild-type strain grown on xylose as carbon source, i.e., when the lowest glucoamylase production rates were observed. A clear negative correlation was found between the oxalic acid production rate and the respiration quotient (RQ). An increase in the oxygen consumption rate, due to the production of strongly oxidized oxalic acid, caused the RQ to be lowest at those stages of recycling cultivation when highest oxalic acid production rates were observed.

We describe a method for measuring urinary oxalate and citrate with capillary electrophoresis (CE) and indirect ultraviolet absorbance detection. Sample preparation is minimal, requiring an acidification, brief centrifugation, and dilution. The method is rapid, with oxalate and citrate having mean migration times of 4.02 and 4.50 min, respectively. The minimal detectable concentration (signal-to-noise ratio of 7) of both oxalate and citrate in urine was 7 mg/L. Total imprecisions (CV) were 1.2-5.6% for three urine samples with oxalate and citrate concentrations of 8-60 mg/L and 80-860 mg/L, respectively. The recovery of added oxalate ranged from 94% to 101%. Results of CE analyses agreed well with enzymatic determinations of oxalate and citrate. Rapid analysis time, accuracy, and reproducibility make this procedure well suited for routine urinary oxalate and citrate determinations.

D-glycero-Pent-2-enono-1,4-lactone (trivial name: D-erythroascorbic acid) occurs in the phytopathogen, Sclerotinia sclerotiorum (Lib.) de Bary, where it has a potential role as precursor of oxalic acid. On Glc/yeast/malt medium, S. sclerotiorum produces only nominal amounts of D-erythroascorbic acid but even partial replacement of Glc by D-Ara increases production of erythroascorbic acid and oxalic acid. Use of D-[1-14C]-, -[3-14C]-, or -[6-14C]Glc and D-[5-3H]-, -[2-14C,5-3H]-, or -[UL-14C]Ara provide additional information on erythroascorbic acid biosynthesis and cleavage. The latter process resembles that obtained by peroxygenation of erythroascorbic acid in alkaline solution. An unknown erythroascorbic acid-like compound also occurs in both Glc- and Ara-based cultures.

The effect of blanching on the antinutritional content was studied in cabbage, turnip, collard, sweetpotato and peanut leaves. All the vegetables contained various amounts of phytic acid, tannic acid and/or oxalic acid. Tannic acid was found in largest amounts ranging from 1266.00 mg/100 g in cabbage to 491.00 mg/100 g in sweetpotato. Phytic acid content ranged from 0.31 mg/100 g in sweetpotato to 3.97 mg/100 g in collard. Oxalic acid was in trace amounts in cabbage and turnip; but high concentrations were found in sweetpotato (469.67 mg/100 g) and peanut greens (407.00 mg/100 g). Levels of both tannic acid and phytic acid were significantly (p < 0.05) reduced by conventional and microwave blanching methods while oxalic acid levels were not significantly (p < 0.05) reduced in most of the treatments by either of the blanching methods. In general, blanching is recommended as an effective method for reducing the antinutritional factors in green vegetables, however, further investigation on the heating times for both conventional and microwave blanching methods has been suggested.

Structural analysis of human kidney stones reveals the presence of cellular membranes and other cell fragments. Experimentally, calcium oxalate crystallization is facilitated when an exogenous nephrotoxin is given with ethylene glycol, thus providing cellular degradation products to act as heterogeneous nuclei. In this report, we tested whether oxalate alone could act as a cell toxin capable of producing damaged cells without the presence of an exogenous agent. Cultured LLC-PK1 and MDCK cells, when exposed to 1.0 mmol KOx, a concentration at the limit of metastability for calcium oxalate nucleation, were severely damaged as measured by specific lactate dehydrogenase (LDH) release in the spent media and by trypan blue exclusion. This effect was magnified by the addition of pre-formed calcium oxalate monohydrate crystals; the injury was significantly amplified when compared to exposure to oxalate alone. Scanning electron microscopy studies illustrated attachment of crystals to cells with loss of cell-to-cell and cell-to-substrate contact, as cells were released from the monolayer. In both oxalate and combined crystal-oxalate studies, more cells were released from the monolayer and exhibited considerably more damage when compared to controls. Oxalate, at the limit of metastability for calcium oxalate, is a cell toxin and can produce cellular degradation products. This effect is increased significantly by the addition of calcium oxalate monohydrate crystals.

Twenty two lines/accessions belonging to 5 species of Celosia were analysed for their nutritional and antinutritional composition. The nutrient contents, vitamin C varied from 18.8 to 53.6 mg/100 g, carotenoids 9.1 to 15.1 mg/100g, protein 2.1 to 5.9%, while the antinutritional factors, nitrate 0.18 to 0.46% and oxalate 0.98 to 3.93%. Moisture ranged 81 to 89%. Variation of these parameters with leaf position and the amino acids profile of some promising lines was also evaluated.

This study addresses two questions: i) which antigens can withstand bleaching by 2.5 g/L of potassium permanganate followed by 10 g/L of oxalate, before immunohistochemical staining; and ii) are any other steps in the immunohistochemical staining technique resistant to bleaching? A panel of 10 antigens was stained immunohistochemically and the results compared with staining performed with a bleaching step interpolated at different steps in the procedures. Four antigens (HMB-45, S-100, factor VIII-related antigen and collagen type IV) were unaffected by bleaching; two antigens (CD-20 and CD-45) had their staining enhanced by bleaching; one had the staining reduced (hsp27); and in three it was abolished (CD-45Ro, CD-31 and Ulex/anti-ulex antibody) by bleaching. Two antibodies (UCHL-1 and L-26) showed evidence for altered specificity following bleaching. None of the steps after application of the primary antibody was resistant to bleaching. Three chromagens used for peroxidase demonstration-amino ethyl-carbazole, diaminobenzidine and chloro-naphthol-were also found to be sensitive to bleaching. While some antigens were resistant to the effects of bleaching, some were not, and no other step in the immunohistochemical procedure could withstand bleaching.