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Sulfite inhibits oxalate production from glycolate and glyoxylate in vitro and from dichloroacetate infused i.v. into male rats.

The effect of sulfite on oxalate production from glycolate and glyoxylate was investigated in an in vitro assay system using rabbit muscle LDH and rat liver 35-60% ammonium sulfate fraction as enzymes. A > or = 50% inhibition in oxalate production was observed at sulfite to glyoxylate ratio of 0.4. LDH activity (change in A340 nm) was inhibited by > or = 40% at sulfite to glyoxylate ratio of 0.5. Male Sprague-Dawley rats fasted for 24 h and infused with dichloroacetate (150 mg/rat/h, iv, n = 6), without and with sodium sulfite (150 mg/rat/h, n = 6) excreted oxalate in urine, respectively, at the rate of 1.13 +/- 0.29 and 0.61 +/- 0.09 mumol/h/100 g body wt, showing a nearly 50% reduction due to sulfite. The present report demands further experimentation to define the role of sulfite as an intracellular modulator of oxalate production.

Alcohol Oxidoreductases↗

Aggregation and dispersion characteristics of calcium oxalate monohydrate: effect of urinary species.

In this research, screening and central composite experimental designs are used to determine the effect of various factors on the aggregation and dispersion characteristics of previously grown calcium oxalate monohydrate (COM) crystals in artificial urinary environments of controlled variables. The variables examined are pH and calcium, oxalate, pyrophosphate, citrate, and protein concentrations in ultrapure water and artificial urine. Optical density measurements, particle size analysis, optical microscopy, AFM force measurements, and protein adsorption have been used to assess the state of aggregation and dispersion of the COM crystals and to elucidate the mechanisms involved in such a complex system. The data indicate that our model protein, mucin, acts as a dispersant. This is attributed to steric hindrance resulting from the adsorbed mucoprotein. Oxalate, however, promotes aggregation. Interesting interactions between protein and oxalate along with protein and citrate are observed. Such interactions (synergistic or antagonistic) are found to depend on the concentrations of these species. Surface responses for these interactions are presented and discussed in this paper. In summary, solution, surface, and interface chemistries interact in a complex manner in the physiological environment to either inhibit or promote aggregation, and an understanding of such interactions may help determine and control the factors affecting kidney stone formation.

Adsorption↗

The effects of oxalate treatment on the smear layer of ground surfaces of human dentine.

The layer was evaluated by scanning electron microscopy and by measurement of hydraulic conductance before and after 2-min topical treatment with potassium chloride, neutral potassium oxalate, half-neutralized oxalic acid or both neutral and acidic oxalates. The treated smear layers were then re-evaluated microscopically and functionally both before and after acid challenge. The layers treated with KCl were not altered either microscopically or functionally and were susceptible to acid etching. Dentine surfaces treated with either oxalate solutions became less permeable and were acid-resistant.

Dentin↗

Subcellular origin of the oxalate- or inorganic phosphate-stimulated Ca2+ transport by smooth muscle microsomes: revisitation of the old problem by a new approach using saponin.

Saponin, a cell-skinning reagent which perforates the cell membrane via its specific interaction with plasmalemmal cholesterol, was used to identify the subcellular origin of ATP-dependent Ca2+ accumulation in the presence and absence of inorganic phosphate and oxalate by microsomal fractions isolated from rat vas deferens and dog aorta. The purified plasma membranes from rat gastric fundus muscle, which elicit the stimulation of ATP-dependent Ca2+ accumulation by inorganic phosphate but not by oxalate, were used as a control reference. Saponin at concentrations effective for skinning smooth muscle fibres (10-50 micrograms/ml) inhibited Ca2+ binding in the absence of ATP to a similar extent in all fractions, but the inhibition of ATP-dependent Ca2+ accumulation was more pronounced in dog aorta microsomes and rat gastric fundus muscle plasma membranes than in rat vas deferens microsomes. The resistance of phosphate- and oxalate-stimulated ATP-dependent Ca2+ accumulation to inhibition by saponin was much greater in rat vas deferens than in dog aorta microsomes. Our results suggest that phosphate- and oxalate-stimulated ATP-dependent Ca2+ accumulation also occurs in plasma membrane vesicles isolated from smooth muscle and is by no means an unique property of endoplasmic reticulum.

Animals↗

Intestinal absorption of oxalate by xylitol-treated rats and mice.

The absorption of 14C-labelled oxalic acid was studied in Wistar rats, CD-1 mice and NMRI mice. Oxalic acid in solution was given to the animals by gavage either with water alone or with 0.625 g/kg b.w. of xylitol. Both xylitol-adapted animals and animals not previously exposed to xylitol were used. Adaptation to xylitol diets enhanced the absorption and urinary excretion of the label (oxalic acid) in both strains of mice but not in rats. Earlier studies have indicated a high incidence of bladder calculi in mice but not in rats fed high amounts of xylitol. The results of the present study offer one likely explanation for the increased formation of bladder calculi as a result of oversaturation of urine with oxalate.

Administration, Oral↗

Simplified method for enzymatic urine oxalate assay.

A procedure for the quantitative determination of oxalate in urine suitable for use in clinical laboratories is described. Oxalate is extracted from urine and subsequently assayed by measuring the amount of hydrogen peroxide produced in an oxidation reaction catalyzed by oxalate oxidase. The reproducibility of the method was assessed by within-run and day-to-day reproducibility studies and the accuracy of the method was assessed by determining the amount of oxalate recovered in spiked urine samples. The correlation obtained between the method described here and a reference method was 0.976.

Adult↗

The inhibition of metabolic oxalate production by sulfhydryl compounds.

A number of sulfhydryl compounds were shown to inhibit CO2 and oxalate formation from glyoxylate by rat liver homogenates and hepatocytes. The most significant inhibition occurred with cysteine and this inhibition was concentration-dependent. In rats made hyperoxaluric by administering ethylene glycol in their drinking water, daily intraperitoneal injections of cysteine caused a rapid and marked decrease in urinary oxalate excretion which was maintained over the duration of the treatment (28 days). Over this time period, the level of urinary oxalate excretion in these ethylene glycol-treated rats was reduced to that of the controls. It is postulated that the decrease is due to the formation of a cysteine-glyoxylate adduct, 2-carboxy-4-thiazolidine carboxylate, which prevents glyoxylate being further oxidized to oxalate. Cysteine or similar sulphydryl compounds may therefore have potential as therapeutic agents in the prevention of renal stones.

Aluminum↗

Binding of oxalate to mitochondrial inner membranes of rat and human kidney.

Oxalate bound specifically to homogenates of rat kidney and liver but not to homogenates prepared from heart, lung, skeletal muscle, spleen, stomach, small or large intestine. In the renal cortex, binding was localized to the inner mitochondrial membrane where it was enriched fourfold when compared to homogenate. Binding of the oxalate reached equilibrium in two minutes at 23C. Analysis of the binding sites by Scatchard plot indicated that the maximum binding capacity was 49 pmol./mg. protein and the apparent dissociation constant (Kd) was 43 nM. The IC50 of oxalate was 0.25 microM. Among the inhibitors studied the IC50 was in the following order: oxalate less than oxamate less than parabanate less than glyoxalate less than oxaloacetate less than malate less than citrate = glycollate. Heat and treatment with lubrol abolished the binding completely. Binding was not enhanced by the presence of calcium in the incubation medium; neither was it inhibited by the presence of calcium together with its transport inhibitors. A binding substance with some characteristics similar to the rat mitochondrial binding factor was also found in the human renal cortex.

Animals↗

Concentration profiles of calcium and oxalate in urine, tubular fluid and renal tissue--some theoretical considerations.

This paper analyzes some aspects of the pathophysiology of urolithiasis. It is emphasized that a better understanding of factors contributing to stone formation can only be gained when the primary nucleation site is identified. Three compartments are considered in which supersaturation as a precondition for stone formation could be present: urine in the urinary tract, tubular fluid from the glomerulus down to the duct of Bellini, and the interstitium of the medulla. From calculations based on micropuncture data it becomes apparent that the oxalate concentration in the tubular fluid at the bend of Henle's loop is 1 or 2 orders of magnitude lower than in the duct of Bellini and that the oxalate concentration maximum invariably must be located in the final urine. The calculation of a tubular concentration profile of oxalate shows, that the probability of intra luminal crystal formation is even less likely for plasma oxalate values of 2-3 microM as compared to 1.2 microM, which therefore should be the correct value. The time necessary for the growth of crystals up to a critical size which can obstruct tubules or ureter is not available in the urinary tract nor in the tubules. However, in the medullary interstitium, where solute concentration is highest, nearly unlimited time for crystal growth is available due to the fact, that in this compartment convective flow is very low. It is concluded that the interstitium of the inner medulla has the best chances to function as the primary nucleation site where particles can be formed of a size which subsequently can obstruct the urinary tract.

Calcium↗

Renal clearance of [14C]oxalate: comparison of constant-infusion with single-injection techniques.

1. The renal clearance of [14C]oxalate was assessed by the constant-infusion technique and single-injection technique (plasma sampling only: one-compartment and two-compartment model; plasma and urine sampling). Healthy volunteers and patients with renal stones were studied. 2. Results with the constant-infusion techniques (with and without urine sampling) were not significantly different from each other. 3. The renal clearance of [14C]oxalate measured with the single-injection technique as compared with the constant-infusion technique was overestimated in the single-injection one-compartment model (52%) as well as in the two-compartment model (30%). 4. The calculated level of plasma oxalate in the healthy volunteers ranged from 1.04 to 1.78 mumol/l (mean 1.39). 5. The biological half-life of [14C]oxalate, estimated by the cumulative excretion of 14C in urine after equilibrium had been established, was 128 min (range: 113-142). 6. The oxalate/creatinine clearance ratio in the healthy volunteers ranged from 1.73 to 2.22 (mean 2.01).

Female↗

EPR spectroscopic characterization of the manganese center and a free radical in the oxalate decarboxylase reaction: identification of a tyrosyl radical during turnover.

Several molecular mechanisms for cleavage of the oxalate carbon-carbon bond by manganese-dependent oxalate decarboxylase have recently been proposed involving high oxidation states of manganese. We have examined the oxalate decarboxylase from Bacillus subtilis by electron paramagnetic resonance in perpendicular and parallel polarization configurations to test for the presence of such species in the resting state and during enzymatic turnover. Simulation and the position of the half-field Mn(II) line suggest a nearly octahedral metal geometry in the resting state. No spectroscopic signature for Mn(III) or Mn(IV) is seen in parallel mode EPR for samples frozen during turnover, consistent either with a large zero-field splitting in the oxidized metal center or undetectable levels of these putative high-valent intermediates in the steady state. A narrow, featureless g = 2.0 species was also observed in perpendicular mode in the presence of substrate, enzyme, and dioxygen. Additional splittings in the signal envelope became apparent when spectra were taken at higher temperatures. Isotopic editing resulted in an altered line shape only when tyrosine residues of the enzyme were specifically deuterated. Spectral processing confirmed multiple splittings with isotopically neutral enzyme that collapsed to a single prominent splitting in the deuterated enzyme. These results are consistent with formation of an enzyme-based tyrosyl radical upon oxalate exposure. Modestly enhanced relaxation relative to abiological tyrosyl radicals was observed, but site-directed mutagenesis indicated that conserved tyrosine residues in the active site do not host the unpaired spin. Potential roles for manganese and a peripheral tyrosyl radical during steady-state turnover are discussed.

Bacillus subtilis↗

X-ray studies on crystalline complexes involving amino acids and peptides. XXXI. Effect of chirality on ionization state, stoichiometry and aggregation in the complexes of oxalic acid with L- and DL-histidine.

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.

Crystallography, X-Ray↗

Effects of sugars and vitamin B-6 deficiency on oxalate synthesis in rats.

Rats fed diets containing galactose as the source of carbohydrate excreted greater amounts of endogenously formed oxalic acid in their urine, compared to rats fed glucose, fructose or sucrose. Rats fed lactose showed similar but less marked effects. The greatest amounts of urinary and fecal oxalate excretions were observed among rats fed galactose and no vitamin B-6. This group had the lowest body weights after 3 weeks of feeding. Control rats fed galactose or lactose diets weighed less than those fed sucrose, glucose or fructose diets. All rats fed galactose developed cataracts. More [14C]oxalic acid was recovered in the urine and kidneys of control rats injected with D-[U-14C]galactose compared to those injected with D-[U-14C]glucose or D-[U-14C]fructose. Similar results were observed in kidneys of vitamin B-6-deficient rats. The possible mechanisms by which galactose and other sugars may be converted to oxalate are discussed.

Animals↗

The clinical significance of assessment of serum calcium oxalate saturation in the hyperoxaluria syndromes.

Estimating calcium oxalate saturation (beta CaOx) in body fluids is proposed as a simple and reproducible procedure to assess the risk of systemic oxalosis in several clinical conditions associated with oxalate retention. beta CaOx was computerized from the measured concentrations of main serum ions. Accurate assay of serum oxalate was crucial for reliability of beta CaOx estimates. However, beta CaOx also depended upon changes of calcium and magnesium concentrations. Patients with end-stage renal failure (ESRF) due to primary or enteric hyperoxaluria had beta CaOx greater than saturation, whereas this happened in only 10 of 25 and two of 24 of those with oxalosis-unrelated ESRF. Bony content of oxalate measured in some of these patients was consistent with these results. In patients with maintained renal function beta CaOx was inversely related to glomerular filtration rate, but the slope was steeper in patients with than in those without hyperoxaluria and beta CaOx reached saturation at earlier stages of renal insufficiency.

Calcium Oxalate↗

Combined liver-kidney transplantation in primary hyperoxaluria type 1. Bone histopathology and oxalate body content.

In three patients with end-stage renal failure due to primary hyperoxaluria type 1, successful combined liver-kidney transplantation enabled us to assess the insoluble oxalate pool, which was compared with the histopathological changes observed in iliac crest biopsy specimens. Good correlation was observed between the histopathological grade of bone oxalosis and the estimated oxalate content of the body. In the end-stage of oxalate bone disease, hyperparathyroidism does not play a significant role in bone resorption, which appears to be the consequence of the granulomatous reaction induced by oxalate deposition. Combined liver-kidney transplantation should be performed long before this stage. Early hepatorenal grafting in uremia secondary to primary hyperoxaluria type 1 would avoid the deleterious clinical consequences of systemic oxalosis and shorten the duration of postransplant hyperoxaluria, which may compromise the course of kidney graft.

Adolescent↗

Effect of vitamin C supplementation on renal oxalate deposits in five-sixths nephrectomized rats.

We have previously reported that hyperoxalemia can be aggravated by vitamin C supplementation in regular hemodialysis patients. The present study was undertaken to examine the validity of this observation in an experimental setting. Fifty five-sixths nephrectomized rats were divided into two groups: 30 rats were allowed free access to water containing 8 mg/ml of vitamin C (100-160 mg/100 g/24 h) and the remainder given tap water without vitamin C. The serum creatinine increased and the Hct decreased gradually; however, there was no difference between the two groups. Plasma vitamin C, oxalate and urinary oxalate levels were higher in the vitamin -treated group than the nontreated rats. Histological examination revealed glomerular and interstitial fibrosis and round cell infiltration as well as tubular cyst formation. Oxalate deposits in renal tubules were found only in vitamin C-treated rats with advanced renal failure. Nontreated animals with equally advanced renal impairment showed no oxalate deposits. These results confirm our previous clinical findings that vitamin C supplementation aggravates the secondary oxalosis of chronic renal failure.

Animals↗

Seasonal variation in urinary excretion of calcium, oxalate, magnesium and phosphate on free and standard mineral diet in men with urolithiasis.

Seasonal variations of urine volume and urinary excretion of calcium, oxalate, magnesium and phosphate were studied in 118 men with urolithiasis on a standard mineral diet and on free diet. The 24-hour urine volume did not vary by season. There was no significant seasonal variation in the excretion of urinary calcium, oxalate, magnesium or phosphate on a standard mineral diet. While on a free diet the patients' urinary excretion of calcium showed statistically significant seasonal differences between age groups. The seasonal variations in the excretion of urinary oxalate and phosphate were statistically significant on free diet, but did not differ by age. The urinary excretion of magnesium was not affected by season nor age. Dietary intake probably best explains seasonal changes in the urinary excretion of calcium, oxalate and phosphate.

Adolescent↗

Effect of vitamin A deficiency on oxalate uptake by rat intestinal brush border membrane vesicles (BBMV) and its contribution towards urolithiasis.

The intestinal uptake of [14C]oxalate, [14C]glyoxylate, and [14C]glycolate are studied in brush border membrane vesicles (BBMV) isolated from vitamin A-deficient and pair-fed control rats. The data obtained indicate that oxalate and its precursors are transported across the BBMV by passive diffusion. The intestinal uptake of glyoxylate and glycolate remains unaltered in vitamin A deficiency, while uptake rate of oxalate was significantly increased (p less than 0.01) in vitamin A-deficient rats as compared to pair-fed controls. In conclusion, the results indicate that vitamin A deficiency leads to hyperabsorption of oxalate through the gut.

Animals↗