Hydroxycitrate: a potential new therapy for calcium urolithiasis.

Alkali supplements are used to treat calcium kidney stones owing to their ability to increase urine citrate excretion which lowers stone risk by inhibiting crystallization and complexing calcium. However, alkali increases urine pH, which may reduce effectiveness for patients with calcium phosphate stones and alkaline urine. Hydroxycitrate is a structural analog of citrate, widely available as an over-the-counter supplement for weight reduction. In vitro studies show hydroxycitrate has the capacity to complex calcium equivalent to that of citrate and that it is an effective inhibitor of calcium oxalate monohydrate crystallization. In fact, hydroxycitrate was shown to dissolve calcium oxalate crystals in supersaturated solution in vitro. Hydroxycitrate is not known to be metabolized by humans, so it would not be expected to alter urine pH, as opposed to citrate therapy. Preliminary studies have shown orally ingested hydroxycitrate is excreted in urine, making it an excellent candidate as a stone therapeutic. In this article, we detail the crystal inhibition activity of hydroxycitrate, review the current knowledge of hydroxycitrate use in humans, and identify gaps in knowledge that require appropriate research studies before hydroxycitrate can be recommended as a therapy for kidney stones.

Calcium lactate as an attractive compound to partly replace salt in blue-veined cheese.

In addition to their high sodium content, cheeses are thought to induce an acid load to the body, which is associated with deleterious effects on consumers' health. Our objective was to explore the use of alkalinizing salts in partial substitution of NaCl to reduce both the sodium content and the acid-forming potential of cheese, without altering its sensory properties. Blue-veined cheeses were produced under industrial conditions, using brine salting followed by dry salting with a 4:1 (wt/wt) mixture of calcium lactate:NaCl or calcium citrate:NaCl. Sodium chloride was used in 2 granulometries: coarse (control treatment) and fine, to obtain homogeneous mixtures with the organic salts. Cheeses were then ripened for 56 d. No major appearance defects were observed during ripening. Calcium lactate substitution decreased the Na content of the cheese core by 33%, and calcium citrate substitution increased the citrate content of the cheese core by 410%, respectively, compared with fine NaCl. This study highlighted the substantial role of salt granulometry in sodium content, with the use of the coarse salt reducing the sodium content by 21% compared with fine salt. Sensory profiles showed nonsignificant differences in bitter and salty perceptions of salt-substituted cheeses with calcium lactate and calcium citrate compared with control cheeses. The use of calcium lactate should be considered to reduce the sodium content and improve the nutritional quality of cheeses while maintaining the sensory quality of the products. Alkalinizing organic salts could replace the acidifying salts KCl or CaCl2, which are currently used in salt replacement and are not recommended for consumers with renal disease. The method described here should be considered by cheese-making producers to improve the nutritional quality of cheese. Additional nutritional optimization strategies are suggested.

Supersaturation of calcium citrate as a mechanism behind enhanced availability of calcium phosphates by presence of citrate.

Dissolution of amorphous calcium phosphate (ACP) in aqueous citrate at varying pH has been studied with perspective of increasing availability of calcium from sidestreams of whey protein, lactose and/or cheese production or on development of new functional foods. ACP formed as an initial precipitate in 0.10 mol L-1 equimolar aqueous calcium chloride, sodium citrate, and sodium hydrogenphosphate was used as model for mineral residues formed during milk processing. Upon acidification of the ACP suspension by hydrochloric acid decreasing pH from 6.5 to 4.5, the transformations of ACP occurred through an 8 h period of supersaturation prior to a slow precipitation of calcium citrate tetrahydrate. This robust supersaturation, which may explain increased availability of calcium phosphates in presence of citrate, presented a degree of supersaturation of 7.1 and was characterized by precipitation rates for 0.10 mol L-1 equimolar aqueous calcium chloride, sodium hydrogencitrate, and sodium hydrogenphosphate with pH 5.5, and for 0.10 mol L-1 equimolar aqueous calcium chloride, sodium hydrogencitrate, and sodium dihydrogenphosphate with pH 4.1, with a degree of supersaturation of 2.7. The crystallization processes were similar according to Avrami's model with a half-life for precipitation of approximately 5 h independent of the degree of supersaturation. Ion speciation based on measurement of pH, and total concentrations of calcium, phosphate and citrate, and of conductivity and calcium ion activity during precipitation indicates a low driving force for precipitation with calcium citrate complex dominating at pH 5.5 and calcium hydrogencitrate complex dominating at pH 4.1. Calcium hydrogencitrate is suggested to be the species involved in the crystal growth followed by solid state transformation to calcium citrate tetrahydrate.

Absorption and Bioavailability of Nano-Size Reduced Calcium Citrate Fortified Milk Powder in Ovariectomized and Ovariectomized-Osteoporosis Rats.

The aim of this study was to evaluate the effects of fortification and nano-size reduction on calcium absorption and bioavailability of milk powder formula in sham, ovariectomized, and ovariectomized-osteoporosis rats as a menopause and menopause-osteoporosis model. Skim milk powder and skim milk powder fortified with calcium citrate and the suitable doses of inulin, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and vitamins D3, K1, and B6 were formulated based on the North American and Western European recommended dietary allowances. Optimization on cycle and pressure of high-pressure homogenizer was done to produce nano-fortified milk powder. In vivo study demonstrated that fortification and calcium citrate nano-fortified milk powder increased absorption and bioavailability of calcium, as well as bone stiffness and bone strength in sham, ovariectomized, and ovariectomized-osteoporosis rats. This study successfully developed an effective fortified milk powder for food application.

Effect of dietary mineral sources and oil content on calcium utilization and kidney calcification in female Fischer rats fed low-protein diets.

We studied the effects of dietary mineral source and oil intake on kidney calcification in 4-wk-old female Fischer rats after consuming the AIN-76 purified diet (AIN-76). A modified AIN-76 mineral mixture was used, although the original calcium (Ca)/phosphorus (P) molar ratio remained unchanged. Rats were fed the modified diets for a period of 40 d before their kidneys were removed on the last day. Ca balance tests were performed on days 31 to 36 and biochemical analysis of urine was also studied. Kidney Ca, P, and magnesium (Mg) in the standard diet group (20% protein and 5% oil) were not affected by the mineral source. Kidney Ca, P, and Mg in the low-protein (10% protein) diet group, were found to be influenced by the dietary oil content and mineral source. In particular, the different mineral sources differentially increased kidney mineral accumulation. Pathological examination of the kidney showed that the degree of kidney calcification was proportional to the dietary oil content in the 10% dietary protein group, reflecting the calcium content of the kidney. The information gathered on mineral sources in this study will help future researchers studying the influence of dietary Ca/P molar ratios, and histological changes in the kidney.

Ca2+-citrate uptake and metabolism in Lactobacillus casei ATCC 334.

The putative citrate metabolic pathway in Lactobacillus casei ATCC 334 consists of the transporter CitH, a proton symporter of the citrate-divalent metal ion family of transporters CitMHS, citrate lyase, and the membrane-bound oxaloacetate decarboxylase complex OAD-ABDH. Resting cells of Lactobacillus casei ATCC 334 metabolized citrate in complex with Ca(2+) and not as free citrate or the Mg(2+)-citrate complex, thereby identifying Ca(2+)-citrate as the substrate of the transporter CitH. The pathway was induced in the presence of Ca(2+) and citrate during growth and repressed by the presence of glucose and of galactose, most likely by a carbon catabolite repression mechanism. The end products of Ca(2+)-citrate metabolism by resting cells of Lb. casei were pyruvate, acetate, and acetoin, demonstrating the activity of the membrane-bound oxaloacetate decarboxylase complex OAD-ABDH. Following pyruvate, the pathway splits into two branches. One branch is the classical citrate fermentation pathway producing acetoin by α-acetolactate synthase and α-acetolactate decarboxylase. The other branch yields acetate, for which the route is still obscure. Ca(2+)-citrate metabolism in a modified MRS medium lacking a carbohydrate did not significantly affect the growth characteristics, and generation of metabolic energy in the form of proton motive force (PMF) was not observed in resting cells. In contrast, carbohydrate/Ca(2+)-citrate cometabolism resulted in a higher biomass yield in batch culture. However, also with these cells, no generation of PMF was associated with Ca(2+)-citrate metabolism. It is concluded that citrate metabolism in Lb. casei is beneficial when it counteracts acidification by carbohydrate metabolism in later growth stages.

A model for calcium permeation into small intestine.

An in vitro model was used to simulate the intestinal permeation of calcium ions depending on the type of salt (carbonate, fumarate, citrate, or gluconate), its concentration (1.0, 2.5, 5.0, or 10 mM/l), and pH (1.3, 4.2, 6.2, or 7.5). To simulate the conditions for calcium permeation in a patient in a fasting state, the solutions were placed in contact with segments of small intestine of pig: stomach, duodenum, jejunum, and ileum. The percent permeation, its rate, and half-time were measured in each case. In all cases, the maximum permeation was seen at 1 mM concentration, depending on pH: 100% for carbonate at pH 1.3; 82% for fumarate, pH 6.2; 79.5% for citrate at pH 4.2, and 81% for gluconate at pH 7.4. The maximum rate of permeation (% h(-1)) was also observed at 1 mM: 2.16 for carbonate at pH 1.3, 0.29 for fumarate at pH 6.2, 0.26 for citrate at pH 4.2, and 0.28 for gluconate at pH 7.4. The shortest half-time permeation (t (1/2), h) for 1 mM solutions depended also on pH (in parentheses): carbonate 0.3 (1.3), fumarate 2.4 (6.2), citrate 2.6 (4.2), and gluconate 2.5 (7.4). The results suggest that calcium carbonate and citrate can be recommended to patients with normal gastric acidity and hyperacidity while fumarate and gluconate to patients with hypoacidity.

An observational study on the effects of nadroparin-based and citrate-based continuous venovenous hemofiltration on calcium metabolism.

BACKGROUND: To study calcium homeostasis during citrate-based compared to nadroparin-based CVVH in critically-ill patients with acute renal failure. METHODS: 11 patients were observed during citrate anticoagulation, 9 with nadroparin and 10 controls. Citrate was chosen for patients with active or at risk for bleeding. RESULTS: The controls had, at 24 h, a median serum iCa of 1.1 mmol/l, the citrate group 0.87 mmol/l and the nadroparin group 1.1 mmol/l (citrate vs. control p = 0.001, citrate vs. nadroparin p = 0.002). At 48 h, iCa was not significantly different anymore. Ca balance was negative for the citrate group in contrast to the nadroparin group (p = 0.012). Median serum PTH was higher (30.0 pmol/l vs. 6.5 pmol/l, p = 0.003) in the citrate group. CONCLUSION: With a relative low target-serum-iCa (0.8-0.9 mmol/l) citrate CVVH-treated patients had a negative daily calcium balance and a temporarily lower iCa level resulting in an enhanced PTH response in comparison to nadroparin.

Functional characterization and Me ion specificity of a Ca-citrate transporter from Enterococcus faecalis.

Secondary transporters of the bacterial CitMHS family transport citrate in complex with a metal ion. Different members of the family are specific for the metal ion in the complex and have been shown to transport Mg(2+)-citrate, Ca(2+)-citrate or Fe(3+)-citrate. The Fe(3+)-citrate transporter of Streptococcus mutans clusters on the phylogenetic tree on a separate branch with a group of transporters found in the phylum Firmicutes which are believed to be involved in anaerobic citrate degradation. We have cloned and characterized the transporter from Enterococcus faecalis EfCitH in this cluster. The gene was functionally expressed in Escherichia coli and studied using right-side-out membrane vesicles. The transporter catalyzes proton-motive-force-driven uptake of the Ca(2+)-citrate complex with an affinity constant of 3.5 microm. Homologous exchange is catalyzed with a higher efficiency than efflux down a concentration gradient. Analysis of the metal ion specificity of EfCitH activity in right-side-out membrane vesicles revealed a specificity that was highly similar to that of the Bacillus subtilis Ca(2+)-citrate transporter in the same family. In spite of the high sequence identity with the S. mutans Fe(3+)-citrate transporter, no transport activity with Fe(3+) (or Fe(2+)) could be detected. The transporter of E. faecalis catalyzes translocation of citrate in complex with Ca(2+), Sr(2+), Mn(2+), Cd(2+) and Pb(2+) and not with Mg(2+), Zn(2+), Ni(2+) and Co(2+). The specificity appears to correlate with the size of the metal ion in the complex.

Comparison of the effects of calcium loading with calcium citrate or calcium carbonate on bone turnover in postmenopausal women.

Calcium supplementation is known to increase bone mineral density and decrease fractures, but the relative efficacy of different forms of calcium supplementation is not established. We compared the effects of calcium carbonate and calcium citrate on markers of bone resorption in older postmenopausal women in an open-labeled crossover study. Forty women were randomized to receive 1000 mg/day of either calcium citrate or calcium carbonate for 12 weeks, followed by a 2-week washout without calcium supplements and 12 weeks treatment with the alternate calcium supplement. All women received vitamin D (900 IU/day). Thirty-four women (25 Caucasian, nine Hispanic) completed the study. No significant differences in the decrease in parathyroid hormone (PTH) or bone specific alkaline phosphatase or the increase in urinary calcium/creatinine were detected between the two treatments. However, calcium citrate supplementation decreased the collagen cross-link resorption markers, urinary N-telopeptide (-30%), C-telopeptide (-31%), free deoxypyridinoline (19%) and serum N-telopeptide (-8%), compared to no significant change following calcium carbonate supplementation (+2%, +3%, +2% and +2%, respectively; P<0.05). Calcium citrate decreased markers of bone resorption significantly more than calcium carbonate in postmenopausal women, although no differences in their effects in calcium excretion or PTH were detected.

Evidence for absorption of ionic calcium and soluble calcium complexes by the duodenum and cecum in the rat.

The absorption of dietary calcium (Ca) may in part be determined by the formation in the intestinal lumen of soluble Ca complexes and insoluble Ca salts. This study was undertaken to test the assumption that ionic Ca concentration (Ca2+) is the only species of Ca that is available for absorption. Bidirectional steady-state Ca fluxes were measured in vitro under short-circuit conditions across segments of the proximal duodenum and the cecum in the presence and absence of varying concentrations of soluble Ca citrate complexes. The presence of 5.0 mmol/L medium citrate reduced medium Ca2+ and cecal Ca mucosal-to-serosal fluxes (Jms) (29 +/- 18 versus 108 +/- 7 nmol Ca/cm2/h, P <.001), but did not reduce duodenal Ca Jms (31 +/- 5 versus 23 +/- 9, P not significant). Duodenal Ca Jms increased 106% as medium Ca citrate complex increased to 1.018 mmol/L and Ca2+ remained constant; cecal Jms increased by 48% under the same conditions. The formation of soluble Ca organic anion complexes with lactate, malate, and fumarate reduced medium Ca2+ and cecal Ca Jms decreased with the reduction of medium Ca2+. The results of this study indicate that Ca2+ is the form of Ca most readily absorbed by the small intestine and the colon. Soluble Ca citrate complexes are absorbed by the duodenum and, to a much lesser extent, by the cecum. The reduction of Ca Jms by citrate is caused by the reduction of medium Ca2+ through formation of Ca citrate complexes and not caused by a direct interaction of the anion with the intestinal epithelium.

Absorption of calcium as the carbonate and citrate salts, with some observations on method.

Calcium supplement use has increased and there is confusion about the relative absorbability of various sources. Absorbability of calcium from the carbonate and citrate salts was compared at 300 mg and 1000 mg calcium loads, ingested as part of a light breakfast meal. Absorption was measured at the high load both by tracer appearance in serum and by the absorptive increment in urinary calcium, and at the low load by the tracer method only. Subjects were 37 healthy adult men and women, studied as outpatients, and each tested on both salts at the same load. Mean tracer absorption (+/- SD) for both salts combined was 36.0% at the 300 mg load and 28.4% at the 1000 mg load. In both experiments the observed mean difference in absorption between salts was very small. By the tracer method the within-subject difference (carbonate less citrate) was +3.3% +/- 1.2% of the ingested dose (mean +/- SEM; P < 0.05) at the high load, and at the low load, 3.6% +/- 2.7% (NS). Combining the two experiments yielded zero difference between sources. By the urinary calcium increment method, the mean difference between salts at the 1000 mg load was 1.8 +/- 4.1 mg (NS). Side-by-side comparisons of the two methods revealed that the tracer method was 3 times more sensitive than the urinary increment method. We conclude that, when taken with food, calcium from the carbonate salt is fully as absorbable as from the citrate, and that the urinary increment method is not sufficiently sensitive to be useful in comparing sources in free-living subjects.

[Dissolution velocity of different calcium preparations used in the clinical field]. / Velocidad de disolución de diferentes preparados de calcio utilizados en clínica.

BACKGROUND: Prescription of calcium supplements is a frequent practice, considering that diet is insufficient to cover daily requirements of this mineral. AIM: To study the dissolution velocity in an acid solution, of different commercial calcium supplements. MATERIAL AND METHODS: Hydrochloric acid was added to distilled water in increasing amounts to obtain a final pH of 6.9, 3.0, 2.5, 2.0 and 1.5. Eighteen commercial calcium preparations were incubated in these solutions for 60 min and dissolution velocity was measured as the percentage of elemental calcium found in solution after this incubation period. RESULTS: Calcium carbonate preparations had a pH dependent dissolution velocity, ranging from 0.67 +/- 0.8% at pH 6.9 to 77.15 +/- 17.5% at pH 1.5. Using the solution with pH 1.5, the dissolution velocity of different preparations varied widely from 56 to 100%. Calcium acetate, followed by calcium citrate and dicalcic phosphate were the salts in tablets with better dissolution velocities. Among powders and effervescent preparations, those containing calcium lactogluconate and citrate had the better dissolution velocities (95 to 115%), that were independent of the solution's pH. A studied preparation with integral bone had a very low dissolution velocity, not surpassing 33 mg of calcium per tablet. CONCLUSIONS: The dissolution velocity of different calcium carbonate preparations varies greatly and, in conditions of achlorhydria, it is negligible. Calcium lactogluconate and citrate dissolution velocities are independent of the solution's pH.

Contribuiçäo do laboratório de análises clínicas no diagnóstico metabólico da litíase renal / The importance of laboratory tests in the metabolic evaluation of nephrolithiasis

Cerca de 5 porcento da populaçäo brasileira é portadora de litíase renal, o que corresponde a cerca de 7 milhöes de litiásicos brasileiros. É uma patologia que acomete indivíduos jovens, geralmente na terceira década de vida, nos seus anos mais produtivos, além de sua recorrência ser muito elevada. Apesar do avanço no tratamento da litíase renal nos últimos anos, através de técnicas näo invasivas, estes métodos só servem para retirar o cálculo, ou seja, eliminam a consequência da doença e näo a sua causa. Desta forma, todo paciente com urolitíase ou que já apresentou cálculo e, principalmente, aquele com história de múltiplos cálculos devem submeter-se a realizaçäo de um estudo metabólico. Com este objetivo, 73 indivíduos com litíase renal recorrente foram estudados. Destes, apenas 1 (1,4 porcento) näo apresentou alteraçäo em nenhum dos parâmetros analisados. Os principais distúrbios metabólicos encontrados foram hipercalciúria, hipernatriúria, hipocitratúria, hipomagnesúria e hiperuricosúria. Este tipo de estudo é importante e o laboratório de análises clínicas contribui na elucidaçäo dos mecanismos fisiopatogênicos responsáveis pela formaçäo de cálculos e na indicaçäo de uma terapêutica mais racional, adequada e eficaz