Solubility of calcium oxalate monohydrate and hydroxyapatite in EDTA solutions

The effect of the addition of a buffer to an ethylenediaminetetracetic acid (EDTA) litholytic solution on the solubility of calcium oxalate monohydrate and hydroxyapatite is investigated. The experiments show that the addition of triethanolamine as a buffer to EDTA solutions at pH 8 and 8.5 enhances the solubility of these salts, confirming theoretical predictions. The solubility of calcium oxalate monohydrate at 25C in solutions with varying ratios of buffer to EDTA concentrations and an osmolality of 0.9 was determined. The solutions resulting in the highest solubility were finally tested for their litholytic ability at 37C. At pH 8 solubilities of 22.5 g X l-1 for calcium oxalate monohydrate and 20.2 g X l-1 for hydroxyapatite were obtained. The corresponding solubilities at pH 8.5 were respectively 24.5 g X l-1 and 19.2 g X l-1.     

Growth retardation of calcium oxalate by sodium copper chlorophyllin

Earlier reports from our laboratories described the inhibitory effect of Mg++ and pyrophosphate on the dissolution and growth of calcium oxalate. In this report, growth of Ca oxalate in presence of very low concentration of chlorophyllin was followed by the Coulter counter and a multichannel analyser. The effect of chlorophyllin concentration on crystal formation was studied at different levels of supersaturation. A concentration of 0.1 microgram/ml produced a remarkable retardation of the induction period and the growth rate. The findings obtained in this study are discussed in the light of the crystal poisoning theory. Comparison was made between chlorophyllin and other growth inhibitors.     

Dietary oxalate and calcium oxalate nephrolithiasis

PURPOSE: Patients with calcium oxalate kidney stones are advised to decrease the consumption of foods that contain oxalate. We hypothesized that a cutback in dietary oxalate would lead to a decrease in the urinary excretion of oxalate and decreased stone recurrence. We tested the hypothesis in an animal model of calcium oxalate nephrolithiasis. MATERIALS AND METHODS: Hydroxy-L-proline (5%), a precursor of oxalate found in collagenous foods, was given with rat chow to male Sprague-Dawley rats. After 42 days rats in group 1 continued on hydroxy-L-proline, while those in group 2 were given chow without added hydroxy-L-proline for the next 21 days. Food and water consumption as well as weight were monitored regularly. Once weekly urine was collected and analyzed for creatinine, calcium, oxalate, lactate dehydrogenase, 8-isoprostane and H(2)O(2). Urinary pH and crystalluria were monitored. Rats were sacrificed at 28, 42 and 63 days, respectively. Renal tissue was examined for crystal deposition by light microscopy. RESULTS: Rats receiving hydroxy-L-proline showed hyperoxaluria, calcium oxalate crystalluria and nephrolithiasis, and by day 42 all contained renal calcium oxalate crystal deposits. Urinary excretion of lactate dehydrogenase, 8-isoprostane and H(2)O(2) increased significantly. After hydroxy-L-proline was discontinued in group 2 there was a significant decrease in urinary oxalate, 8-isoprostane and H(2)O(2). Half of the group 2 rats appeared to be crystal-free. CONCLUSIONS: Dietary sources of oxalate can induce hyperoxaluria and crystal deposition in the kidneys with associated degradation in renal biology. Eliminating oxalate from the diet decreases not only urinary oxalate, but also calcium oxalate crystal deposits in the kidneys and improves their function.     

Kinetics of dissolution of calcium oxalate calculi with calcium-chelating irrigating solutions

Dissolution of calcium oxalate urinary calculi was studied in vitro. Rate constants were calculated by measuring the concentration of calcium in solution during dissolution. Various irrigating agents, including ethylenediaminetetraacetic acid (EDTA) and other calcium-chelating substances, were tested at different concentrations and pH values. EDTA was the most effective agent in dissolving calcium oxalate calculi. When EDTA was used, the rate of dissolution depended on both concentration and pH. At a pH of 7.5 or 10.0, the rate of dissolution increased progressively with the EDTA concentration. At each concentration tested, dissolution was faster at a pH of 10.0 than at a pH of 7.5. Calculated rate constants were extrapolated to determine whether in vivo irrigation with EDTA solutions is clinically practical. Even at an EDTA concentration of 0.03 M at a pH of 7.5, a two mm. calculus could be dissolved within 48 hr. Although dissolution of calcium oxalate calculi is not a practical first-line treatment, it might be a useful adjunct to percutaneous stone removal or extracorporeal shock wave lithotripsy.     

一水草酸钙与二水草酸钙结石形成机理的研究

目的　探讨一水草酸钙（ＣＯＭ） 与二水草酸钙（ＣＯＤ） 结石形成的机制。　方法　应用红外光谱仪对２５８ 块尿结石成分进行检测,同时检测３０ 例患者２４ｈ 尿液生化指标,对测定结果利用ＳＰＳＳ软件进行ｔ 检验。　结果　（１） 尿钙：ＣＯＭ 组（４．８３ ±１ ．９８）ｍ ｍｏｌ／２４ｈ,ＣＯＤ 组（９．８８ ±４ ．２８）ｍｍｏｌ／２４ｈ,Ｐ＜ ０ ．０１ ;（２） 尿磷：ＣＯＭ 组（１９ ．４０ ±９．６９）ｍ ｍｏｌ／２４ｈ,ＣＯＤ 组（２９．２０ ±１２．００）ｍ ｍｏｌ／２４ｈ,Ｐ＜ ０．０５,两组尿钙、尿磷差异有显著性。　结论　二水草酸钙结石患者尿钙、尿磷高于一水草酸钙结石患者,表明二水草酸钙的形成与高钙尿及磷酸盐异质成核有关,而一水草酸钙的形成可能与尿中抑制物缺乏有关。     

Crystallisation of calcium oxalate dihydrate in normal urine in presence of sodium copper chlorophyllin

Summary A method is described for the growth of calcium oxalate dihydrate in normal urine. Soluble chlorophyllin, at a concentration of 20 g/ml inhibited the crystallisation and the growth kinetics of the dihydrate crystals. The inhibitory capacity of chlorophyllin was compared with previous results. Data obtained suggest that the food and drug colourant chlorophyllin might be useful in the treatment of calcium oxalate stone disease.     

Plasma oxalate concentration in calcium oxalate stone formers

A sensitive, simplified method for plasma oxalate determination by gas chromatography is described. After deproteinizing the plasma with 3N HC1 and 20% sulfosalicylic acid, the oxalate was methylated, extracted and analysed by gas chromatography. This method has three advantages i.e., smaller sample size (plasma 5.0 ml), rapidity (takes less than 3 hours) and accuracy. The recovery rate of oxalate added to plasma was 91.42 +/- 11.31% (SD) and the coefficient of variation of replicate determinations was 4.18%. The minimum detectable concentration of oxalate was 0.3 micrograms/ml (oxalate peak was higher than 5 mm). The mean oxalate concentration under fasting conditions from 16 healthy subjects was 1.37 +/- 0.39 micrograms/ml (SD), while that from 31 calcium oxalate stone formers was 1.45 +/- 0.39 micrograms/ml (SD). There was no significant difference in plasma oxalate concentration between the two groups. The dietary influence of oxalate on plasma and urinary oxalate was investigated in 5 healthy subjects and 5 calcium oxalate stone formers. When 100 g spinach (total oxalate 545.5 mg, soluble oxalate 381.5 mg) was given, the increase of plasma oxalate concentration was more prominent in stone formers; in stone formers it increased to 142% of control value at 2 hours (p less than 0.05) after spinach loading, to 163% at 4 hour (p less than 0.01) and to 232% at 6 hours (p less than 0.01); while in healthy subjects increased to 119% at 2 hours (ns) after loading, to 144% at 4 hours (p less than 0.05) and only to 167% at 6 hours (p less than 0.01). Urinary oxalate excretion increased promptly between 1 and 2 hours after loading in both groups, reaching peak levels between 2 and 4 hours after loading in healthy subjects and between 4 and 6 hours or later in stone formers. The mean renal clearance of oxalate was 18.0 ml/min in 6 healthy subjects and 19.0 ml/min in 4 calcium oxalate stone formers. There was no significant difference in oxalate clearance between the two groups. The mean ratio of oxalate/creatinine clearance was 0.22 for stone formers, which was equal to that for healthy subjects.     

Role of magnesium in the growth of calcium oxalate monohydrate and calcium oxalate dihydrate crystals

Since about 85% of synthesized calcium oxalate dihydrate (COD) crystals proved not to have changed into calcium oxalate monohydrate (COM) crystals at 30 min of incubation time at 37 degrees C when our evaluation method of the COD-to-COM ratio was being used, we made a comparative study of the inhibitory effects of magnesium, one of the well-known inhibitors of calcium oxalate stone formation, on the growth of seeded COM and COD crystals. The results demonstrated that magnesium in identical concentrations might have stronger inhibitory effects on the growth of COM crystals than on that of COD crystals and suggested that these different effects of magnesium on the growth of COM and COD crystals might arise not only from the difference between the specific surface areas of COM and COD crystals, but also from that between the direct inhibitory effects of magnesium on these two types of calcium oxalate crystal growth.     

正常人和草酸钙结石病人尿草酸钙晶体基质

以改良Ｍｏｒｓｅ和Ｒｅｓｎｉｃｋ法提取１０例上尿路草酸钙结石病人和１１例正常人的尿草酸钙晶体基璺，用双向聚丙烯酰胺凝胶电泳对晶体基质及结晶前后大分子物的蛋白质组成进行了比较分析。     

Effects of sodium urate and uric acid crystals on the crystallization of calcium oxalate

Summary Crystallization of calcium oxalate in the presence of uric acid and sodium urate crystals was analyzed in a metastable crystallization system containing calcium chloride and sodium oxalate (A), in urine highly supersaturated with respect to calcium oxalate (B), and in urine with a high level of metastable supersaturations (C). In system A uric acid crystals in concentrations up to 11.4 mMol/l did not affect calcium oxalate crystallization, neither did sodium urate during the first 6 h in concentrations below 5 mMol/l. In system B neither uric acid nor sodium urate crystals affected calcium oxalate crystallization. However, an increased rate of crystallization was observed with both uric acid and sodium urate in system C, but the effect was less pronounced than with calcium oxalate seed. Urine pre-treated with sodium urate and subsequently analyzed in system A in a concentration of 2%, gave a slightly lower inhibition of calcium oxalate crystal growth. Concerning the crystal size distribution in the same system, larger crystals were observed in several urines pre-treated with uric acid and sodium urate.     

Relationship between supersaturation and calcium oxalate crystallization in normals and idiopathic calcium oxalate stone formers

BACKGROUND: In an earlier study on recurrent CaOx stone formers with no detectable abnormalities, we found that the urine of these subjects had a lower tolerance to oxalate load than controls and that the removal of urinary macromolecules with a molecular weight greater than 10,000 D improved their tolerance to oxalate. METHODS: The effects on CaOx crystallization of reduced urinary supersaturation of calcium oxalate (CaOx), induced by night water load, were studied in 12 normal males and in 15 male OxCa stone formers who were free from urinary metabolic abnormalities. The effect of the macromolecules, purified and retrieved from the natural and diluted urine, were analyzed in a metastable solution of CaOx. RESULTS: The water load caused an increase in urine volume (from 307 +/- 111 to 572 +/- 322 ml/8 hr, P = 0.014 in normal subjects, and from 266 +/- 92 to 518 +/- 208 ml/8 hr, P = 0.001 in the stone formers) and a concomitant reduction of the relative CaOx supersaturation (from 8.7 +/- 2.5 to 5.1 +/- 2.5 ml/8 hr, P = 0.001 in normal subjects, and from 10.4 +/- 3.5 to 5.0 +/- 2.7 ml/8 hr, P = 0.001 in the stone formers). The decrease in CaOx supersaturation was accompanied by an increase of the permissible increment in oxalate, both in normal subjects (from 43.8 +/- 10.1 to 67.2 +/- 30. 3 mg/liter, P = 0.018) and in the stone formers (from 25.7 +/- 9.4 to 43.7 +/- 17.1 mg/liter, P = 0.0001), without any significant variations of the upper limit of metastability for CaOx (from 21.6 +/- 5.3 to 20.5 +/- 4.2 mg/liter in normal subjects, and from 18.7 +/- 4.5 to 17.1 +/- 3.7 mg/liter in the stone formers). The inhibitory effect of urinary macromolecules with molecular weight greater than 10,000 Daltons did not undergo any change when the latter were recovered from concentrated or diluted urine, either in normal subjects or in the stone formers. CONCLUSIONS: Reduced CaOx supersaturation by means of water load has a protective effect with regards to CaOx crystallization in subjects who do not present any of the common urinary stone risk factors.     

草酸钙晶体表面结合蛋白质的研究

目的 了解草酸钙晶体表面结合蛋白质在结石形成的过程中的作用。方法 用草酸钙过饱和结晶法制备正常人和草权钙结石患者尿草酸钙晶体表面结合物质（ＣＳＢＳ）,经ＤＥＡＥ－ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ柱层析分离蛋白质和葡胺聚糖,用ＳＤＳ－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）测定蛋白质组成和分子量,用氨基酸自动分析仪测定蛋白质的氨基酸,结果;正常仍ＣＳＢＳ中主要含分子量为３１０００的尿凝血酶原激活肽片段１（     

Rapid Communication: relative effect of urinary calcium and oxalate on saturation of calcium oxalate

BACKGROUND: The study compared the effect of urinary calcium with that of oxalate on urinary saturation [relative saturation ratio (RSR)] of calcium oxalate. METHODS: A retrospective data analysis was conducted on urinary stone risk analysis from 667 patients with predominantly calcium oxalate stones. Urinary RSR of calcium oxalate was individually calculated using Equil 2. A "theoretical" curve of the relationship between urinary RSR of calcium oxalate and concentration of calcium or oxalate was obtained at two stability constants for calcium oxalate complex, while varying calcium or oxalate and using group mean values for urinary constituents. RESULTS: At the stability constant of 7.07 x 10(3), the increase in RSR of calcium oxalate was less marked with calcium than with oxalate. However, at the stability constant of 2.746 x 10(3) from the Equil 2 that is considered the "gold standard," calcium and oxalate were equally effective in increasing RSR of calcium oxalate. The above theoretical curves (relating RSR with calcium or oxalate) were closely approximated by the actual curves constructed with data from individual urine samples. Urinary saturation of calcium oxalate was equally dependent on urinary concentrations of calcium and oxalate (r= 0.75 unadjusted and 0.57 adjusted for variables, and P < 0.0001 for calcium; r= 0.73 unadjusted and 0.60 adjusted, P <0.0001 for oxalate). CONCLUSION: Among calcium oxalate stone-formers, urinary calcium is equally effective as urinary oxalate in increasing RSR of calcium oxalate.     

Uric acid-binding proteins in calcium oxalate stone formers and their effect on calcium oxalate crystallization.

OBJECTIVES: To study the effect of urinary uric acid-binding proteins of controls and stone formers on calcium oxalate crystal nucleation and aggregation. MATERIALS AND METHODS: Urine samples were collected over 24 h from 20 stone formers and from 20 age-matched normal controls. Uric acid crystallization was induced by adding equal volumes of 2.5 mmol/L uric acid. The bound proteins were separated on a cellulose column, and by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The effect of the separated fractions on calcium oxalate crystal nucleation and aggregation was assessed. RESULTS: The protein bound to unit mass of uric acid crystals was higher in hyperoxaluric urine than in control urine. On cellulose-column separation, the uric acid-crystal binding proteins produced three major protein peaks, i.e. fraction I (buffer), fraction II (0.05 mol/L sodium chloride in Tris-HCl buffer) and fraction III (0.3 mol/L sodium chloride in buffer), with a minor peak obtained on elution with increasing concentrations of sodium chloride in Tris-HCl buffer (pH 7.0). Fraction I derived from either stone formers or controls promoted calcium oxalate crystallization. Fraction II from the control samples was a strong inhibitor, whereas hyperoxaluric fraction II was less inhibitory. CONCLUSION: Uric acid-binding proteins isolated either from the urine of stone formers or controls modulated calcium oxalate crystal growth. Proteins isolated from stone formers were less inhibitory of crystal nucleation and aggregation. These proteins may act as a bridge, leading to the epitaxial deposition of calcium oxalate over a urate core.     

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.     

Bariatric surgery, calcium oxalate urinary stones and oxalate nephropathy

Bariatric surgery is now recognized as a sure and effective way for weight reduction in morbid obesity. However some procedures induce intestinal malabsorption leading to enteric hyperoxaluria. So bariatric surgery could place these patients not only at risk for nephrolithiasis but also for oxalate induced nephropathy and chronic renal failure. Because of the growing incidence of obesity worldwide, physicians and patients should be aware of such potential complications. There is no mean to discuss this treatment because of its spectacular efficiency on obesity and its comorbidities. But it is necessary to choose the surgical technique according to the risk factors of the patients. Following surgery, preventive treatment strategies are indicated, such as modified dietary lifestyle and specific drugs as we suggested to limit or even avoid these complications. However observance could fail in the long term. In case of oxalate nephropathy, surgery may be proposed to restore the intestinal tract but with the risk of overweight relapse. To illustrate this matter, we report here significant observations of three patients, which, having successfully benefited from the same bariatric surgery, have presented lithiasic complications for two of them and oxalate nephropathy leading to chronic renal failure and hemodialysis for the third.     

Urinary calcium and oxalate excretion in children

We have established normal values for calcium/creatinine (Ca/Cr) and oxalate/creatinine (Ox/Cr) ratios in 25 infants (aged 1–7 days) and 391 children (aged 1 month to 14.5 years) and compared these with values obtained in 137 children with post-glomerular haematuria and 27 with nephrolithiasis. Oxalate was measured by ion chromatography. Nomograms of Marshall and Robertson were used to calculate urine saturation to calcium oxalate. The Ca/Cr ratio was normally distributed whereas the Ox/Cr ratio had a log-normal distribution. The molar ratio of Ca/Cr was the lowest in the first days of life and the highest between 7 month and 1.5 years (mean±SD=0.39±0.28 mmol/mmol). Following a slight decrease it stabilised by the age of 6 years (0.34±0.19 mmol/mmol). The highest Ox/Cr values were measured during the 1st month of life [geometric mean 133 (range 61–280) mol/mmol], followed by a gradual decrease until 11 years of age [mean 24 (range 6–82) mol/mmol]. Thirty-six haematuric children had hypercalciuria (26%), 23 had absorptive hypercalciuria, 13 renal type. Children with absorptive hypercalciuria on a calcium-restricted diet had significantly higher oxalate excretion than those with renal hypercalciuria and the control group [38 (range 28–49) vs. 22 (range 16–29) and 23 (range 22–27) mol/mol respectively,P<0.01]. Calcium oxalate urine saturation of stone patients was higher than that of patients with haematuria and the normal population (1.18±0.05 vs. 1.06±0.03,P<0.03 and 0.84±0.03,P<0.001 respectively). The measurement of Ca/Cr and Ox/Cr in first-morning urine samples is suitable for screening for hypercalciuria and hyperoxaluria. Interpretation of the values requires age-specific reference values. Both calcium and oxalate determinations should be part of the evaluation of patients with haematuria, hypercalciuria or nephrolithiasis.