The surgeon's role on chemical investigations of the composition of urinary stones.

The chemical analysis of an urolith is often interpreted as "stone's composition". However, it must be taken into consideration, that in most cases, only a fragment of the stone has been sent to the laboratory. In some recurrent patients, stone compositions either vary considerably between episodes or the analytical result obtained from the stone fragment does not fit with the data of e.g. current 24 h-urinalysis or urinary pH-records. The question arises, whether this outcome may be the result of an improper stone sampling scheme. On a simple layered 2D-stone model composed of two mineral phases it is shown, how the choice of a stone fragment process may influence the result of "stone composition". Depending on the initial position of fragment within the whole stone, the respective calculated analyses can relevantly differ from the whole stone composition as well as strongly between two fragments. Even under the simplified conditions of a 2D-2-component-model "grown" under defined conditions, the differences between the analyses of the different specimens taken from a stone are in part remarkable. The more it can be argued that these differences increase if a real 3D-urolith is investigated. Further sampling biases may evolve and increase the problem of proper sampling:, e.g., if an urolith's more resistant parts remain intact while ESWL or laser-based stone fragmentation ("dusting"), the weak parts became fully disintegrated and removed from the body as fine-grained sludge-the stone's fine fraction is lost although its composition may carry important information on the stone's pathogenesis. Consequently, a "stone analysis" only obtained from the harder remains reveals an incomplete result, a fact that in principle limits its clinical interpretation. Choice of stone fragment is crucial. The extent of the uncertainty of an analysis resulting from potential selection biases should not be underestimated. Thus, sampling should be considered as an important part of the processes of quality assurance and management. Errors made at this early stage of diagnosis finding will affect the analytical result and thus influence the clarification of the underlying pathomechanism. This can lead to an improper metaphylactic strategy potentially causing recurrent stone formation which otherwise would have been prevented. A decision scheme for analysis of urinary stones removed using endoscopic methods is suggested.

Kinetics of calcium oxalate crystal formation in urine.

It is routinely observed that persons with increased urinary stone risk factors do not necessarily form uroliths. Furthermore, stone formers can present with urinalyses that do not reflect the clinical picture. We explain this discrepancy by differences in crystallization kinetics. In 1162 urines, crystallization of Ca-oxalate was induced according to the BONN-Risk-Index (BRI) method. The urine's relative light transmissivity (RLT) was recorded from 100 % at start of titration to 95 % due to nuclei formation and crystal growth. From the RLT changes, a measure of the thermodynamic inhibition threshold of crystal formation (BRI) and of crystal growth kinetics is derived ("turbidity slope" after crystallization onset). On average, subjects presenting with a low inhibition threshold, i.e., high BRI, also present significantly higher crystal growth rates compared with subjects in lower BRI classes. Only subjects in the highest BRI class show a lower growth rate than expected, probably due to a depletion of supersaturation by massive initial nucleation. With increasing thermodynamic risk of crystal formation (i.e., increasing BRI) due to an imbalance between inhibitors and promoters of crystal formation, an increase in the imbalance between inhibitors and promoters of crystal growth (i.e., increasing growth rate) is observed. Both lead to an increased urolith formation risk. Healthy subjects with increased BRI are an exception to this trend: their urine is thermodynamically prone to form stones, but they show a kinetic inhibition preventing nuclei from significant growth.

Hydrophobic forces as a key factor in crystalline biofilm formation on ureteral stents.

BACKGROUND: Current discussions about biofilm formation focus on the solid/liquid interface between a medical device and body fluids. Yet it has been shown that gas bubbles (GB) can stably form on ureteral stents in artificial urine and that their fate depends on the stent's surface properties. The liquid/gas interface constitutes an adhesion site for precipitating salts as well as hydrophobic organic molecules. MATERIALS AND METHODS: The surface wettability of polyurethane stents is varied by coating with amorphous hydrogenated carbon (a-C:H). GB and crystalline biofilm formation on the stents are investigated in a novel encrustation device which avoids gravitation- or sample-position-related influences on the results. RESULTS: Bigger and more stable GB form on hydrophobic stents than on hydrophilic, coated stents. Appearance and amount of crystalline deposits differ significantly between the surfaces. With decreasing wettability the number of hollow crystalline spheres and the mass of precipitate increase. CONCLUSIONS: On hydrophobic surfaces, stable GB increase precipitation of salts and become incorporated in the growing encrustation layer in vitro. In contrast, GB quickly lift off from hydrophilic surfaces taking part of the precipitate with them. This self-cleaning mechanism slows down the encrustation process. A similar effect may explain the prolonged complication-free indwelling time of amorphous-carbon coated stents in vivo.

Ureteral stents should be soaked for several minutes before placement.

PURPOSE: Placement of ureteral stents (DJ-stents) may lead to complications. Inappropriate friction properties of the implant are, inter alia, made responsible for primary injuries, injury-related inflammation and a cascade of consecutive side effects. Hydrophilicity is considered to be related to low friction. The question arises, whether the various products on the market show their respective maximum hydrophilicity directly after unwrapping or a pre-use moistening, as already routinely done with the guide wire, is necessary. METHODS: The surface wettability of commercial and experimental DJ-stents was determined by water contact angle (WCA) measurements using the sessile drop method. One reference surface and 11 different stent surface types were tested. In order to determine the influence of moistening on the stents' surface wettability, WCAs were measured twice, with dry, and soaked (30 min, 0.9%-NaCl) specimens. Each sample of a surface type was tested at three different positions to avoid effects of surface heterogeneities. Up to six samples of the same surface type were examined. RESULTS: Mean WCAs on fresh and soaked stent surfaces ranged from 75°-103° and 71°-99°. In every case the WCAs on soaked surfaces were lower. On average the WCAs decrease by 7%, the individual decreases differ considerably, from 2 to 16%. For 7/12 of the examined surface types, the decrease in contact angle is statistically significant with p ≤ 0.01. CONCLUSIONS: DJ-stents freshly unwrapped show less hydrophilic properties compared to DJ-stents soaked in saline. To obtain maximum hydrophilicity at stent placement, DJ-stents should be soaked. The results may advocate a similar approach for other urological equipment.

Urolithiasis--an interdisciplinary diagnostic, therapeutic and secondary preventive challenge.

BACKGROUND: The prevalence of urolithiasis in Germany is 4.7%; its incidence has trebled in the last three decades. The risk of recurrence is 50-80%, depending on the type of stone, unless secondary prevention is instituted. Risk-adapted secondary prevention lowers this risk to 10-15%. METHODS: This review is based on publications retrieved by a selective search in PubMed using the key words "urolithiasis," "urinary stones," "epidemiology," "lithogenesis," "biominerals," "risk factors," and "diagnosis, therapy, metaphylaxis." These publications were evaluated with the aid of the urolithiasis guideline of the European Association of Urology. RESULTS: Acute renal colic can usually be diagnosed without sophisticated equipment. Stones can be dealt with by a variety of techniques depending on their size and location, including extracorporeal shock-wave lithotripsy, ureterorenoscopy, percutaneous nephrolitholapaxy, and open surgery. Most ureteric stones of diameter up to 5 mm pass spontaneously. 75% of patients have no complications. The basic evaluation needed for secondary prevention can be carried out by any physician on an ambulatory basis. In the 25% of patients who have complications, a more extensive interdisciplinary evaluation of metabolic parameters should be performed in a clinical center for urinary stones. CONCLUSION: Urolithiasis has many causes and can be treated in many different ways. An extensive metabolic work-up is often necessary for secondary prevention. The various treatment options must be considered for their suitability in each individual patient. Robust data are now available on surgical and interventional methods, but there are as yet no high-quality trials of secondary prevention. Further research should concentrate on the etiology and pathogenesis of urolithiasis.

Induced urinary crystal formation as an analytical strategy for the prediction and monitoring of urolithiasis and other metabolism-related disorders.

Crystal formation reflects the entire composition of the surrounding solution. In case of urolithiasis, induced crystal formation in native urine has led to the development of the Bonn-Risk-Index (BRI), a valuable tool to quantify an individual's risk of calcium oxalate urolithiasis. If the progression of a disease is associated with characteristic changes in the activities of urinary components, this leads to an altered urinary crystallisation capacity. Therefore, the results of induced urinary crystal formation can be used to detect and monitor any disease linked to the altered urinary composition. Since crystal formation inherently takes into account the entire urinary composition, the influence of the disease on individual urinary parameters does not have to be known in order to monitor the consequent pathologic alterations. In this paper, we review the background of urinary crystal formation analysis and describe its established application in urolithiasis monitoring as well as potential further fields of clinical application.

Correspondence between Ca²âº and calciuria, citrate level and pH of urine in pediatric urolithiasis.

BACKGROUND: Hypercalciuria and hypocitraturia are considered the most important risk factors for urolithiasis. Citrate binds to urinary calcium to form a soluble complex which decreases the availability of ionized calcium (Ca(2+)) necessary for calcium oxalate formation and phosphate crystallization. The aims of this study were to assess the Ca(2+) fraction in relation to total calciuria, citraturia and urinary pH and to determine whether urinary Ca(2+) concentration is a helpful biomarker in metabolic evaluation of children with urolithiasis. METHODS: We collected 24-h urine samples from 123 stone-forming children and adolescents with hypocitraturia and from 424 healthy controls. Total calciuria (total calcium, Catotal), Ca(2+), pH, citrate, oxalate and Bonn Risk Index (BRI) were assessed and compared between the two groups. RESULTS: Total calciuria and Ca(2+) content were higher in stone-formers than in the healthy children. In both stone-formers and controls, Ca(2+) content was inversely related to citraturia and urinary pH, whereas the Ca(2+)/Catotal ratio differed slightly between the groups. A large variability in Ca(2+) level was found across individuals in both groups. The BRI increased with increasing calciuria and urine acidity. CONCLUSIONS: Compared to controls, stone-formers with hypocitraturia demonstrated a higher urinary Ca(2+) concentration, but this was proportional to calciuria. The large individual variability in urinary Ca(2+) content limits its practical use in metabolic evaluation of children with urolithiasis. However, the Ca/Citrate ratio may be a useful clinical tool in evaluating children with urolithiasis.

Assessment of lithogenic risk in children based on a morning spot urine sample.

PURPOSE: The Bonn Risk Index has been used to evaluate the risk of urinary calcium oxalate stone formation. According to the original method, risk should be determined based on 24-hour urine collection. We studied whether the Bonn Risk Index could be measured in spot urine samples and which part of the day is most suitable for this purpose. MATERIALS AND METHODS: We collected total and fractionated 24-hour urine (in a 6-hour nocturnal portion and 9 consecutive 2-hour diurnal samples) in 42 children and adolescents with calcium oxalate urolithiasis and 46 controls. Bonn Risk Index values determined from each of the urine fractions were compared to those obtained from related 24-hour urine collections. RESULTS: Both groups exhibited similar circadian patterns of Bonn Risk Index values. Median Bonn Risk Index for the nighttime portion of urine in the stone group was 1.4 times higher than that obtained from the total 24-hour urine. The morning hours between 08:00 and 10:00 showed the peak lithogenic risk, and this fraction had the highest sensitivity and selectivity regarding discrimination between stone formers and healthy subjects. The afternoon hours demonstrated lower and less fluctuating crystallization risk. Despite diurnal fluctuations in Bonn Risk Index, there was still a well-defined cutoff between the groups. CONCLUSIONS: Bonn Risk Index determined from urine samples collected between 08:00 and 10:00 appears optimal in separating stone formers from healthy subjects, and appears as useful as the value determined from 24-hour urine collection. Investigation of this diurnal sample simplifies diagnosis in pediatric stone disease without loss of clinical information.

Metabolic stress response patterns in urinary compositions of idiopathic calcium oxalate stone formers, patients with chronic bowel diseases and controls.

Emotional stress is associated with e.g. increased hormone release, high blood-sugar level and blood pressure. Stress clearly affects metabolism. Whether chronic stress exposure leads to altered urinary compositions with increased risk of CaOx; urolithiasis was examined by investigating the relation between stress burden and urine composition. 29 controls (CG), 29 CaOx stone formers (SF), and 28 patients with chronic inflammatory bowel diseases (CIBD) were advised to avoid unfavorable aliment. Any urolithiasis-related medications were stopped. At day 5, a 24-h urine was collected and comprehensive urinalysis performed. AP (CaOx) index was calculated. Subjects completed a questionnaire designed to measure perceived stress ("Trier-Inventory-of-Chronic-Stress"). Mean AP (CaOx) in CG, SF and CIBD amount to 0.8 (+/-0.3), 1.2 (+/-0.7), and 1.9 (+/-1.2), respectively. Increased AP (CaOx) in SF is mainly attributed to an increased effect of calcium and oxalate, whereas in CIBD this is additionally caused by a reduced effect of citrate, magnesium and volume. Stress dimensions are correlated to any investigated urinary parameter with an absolute value of r < or = 0.600; some correlations are statistically significant: whereas in SF only one combination, "lack of social recognition" versus calcium, shows significance, in CIBD various combinations are significantly related. In particular, sodium excretion increases with stress. In CG, some stress dimensions are directly related to citrate; with increasing stress, protection against CaOx crystallization tends to increase. It could be shown that stress load and urinary composition are related by statistical means. The observed metabolic stress response patterns in urinary compositions are different for the distinct groups, thereby, reflecting a conclusive picture. This is in particular in CIBD, for which a link between stress and inflammatory activity and between inflammatory activity and altered urinary composition is well established.

Relevance of the BONN Risk Index for metabolic monitoring of patients with calcium oxalate urolithiasis: a clinical application study of the Urolizer.

The BONN Risk Index (BRI) successfully determines the calcium oxalate (CaOx) crystallization risk from urine samples. The BRI is based on a standardized crystallization test performed on native urine. A BRI-measuring device, the "Urolizer", has been developed, operating automatically and requiring only a minimum of preparative efforts. In this study, the Urolizer is evaluated regarding its analytical and diagnostic practicability for metaphylaxis control in the framework of the daily routine of a stone surgery. From 51 CaOx recurrent stone-formers, 24 h urines were collected at the beginning and after 3 months of metaphylaxis. As much as 27 patients were indicated to suffer from "mild hypercalciuria", low urinary pH or hypocitraturia, and 24 patients from "hypercalciuria". The former were treated with alkaline citrate (AC), and the latter with hydrochlorothiazide (HCT). Analyses of urines collected before and during treatment, BRI using the Urolizer, and urinalysis-based risk indices were evaluated. In both patient groups, BRI decreased significantly, while metaphylaxis (P<0.001) in the AC group decreased from 1.08 (+/-0.58) to 0.56 (+/-0.39) L(-1) and in the HCT-group from 3.30 (+/-1.15) to 1.60 (+/-0.52) L(-1). In most patients, urinary parameters changed as desired and related risk indices decreased appropriately. The clinical utility of the easy-to-determine BRI is demonstrated. By quantifying the "overall" therapy effect within 15 min, the innovative analysis device may be especially suited for practitioners specializing in urolithiasis treatment.

Analytical precision of the Urolizer for the determination of the BONN-Risk-Index (BRI) for calcium oxalate urolithiasis and evaluation of the influence of 24-h urine storage at moderate temperatures on BRI.

BACKGROUND: Since its first publication in 2000, the BONN-Risk-Index (BRI) has been successfully used to determine the calcium oxalate (CaOx) crystallization risk from urine samples. To date, a BRI-measuring device, the "Urolizer", has been developed, operating automatically and requiring only a minimum of preparation. Two major objectives were pursued: determination of Urolizer precision, and determination of the influence of 24-h urine storage at moderate temperatures on BRI. METHODS: 24-h urine samples from 52 CaOx stone-formers were collected. A total of 37 urine samples were used for the investigation of Urolizer precision by performing six independent BRI determinations in series. In total, 30 samples were taken for additional investigation of urine storability. Each sample was measured thrice: directly after collection, after 24-h storage at T=21 degrees C, and after 24-h cooling at T=4 degrees C. Outcomes were statistically tested for identity with regard to the immediately obtained results. RESULTS: Repeat measurements for evaluation of Urolizer precision revealed statistical identity of data (p-0.05). 24-h storage of urine at both tested temperatures did not significantly affect BRI (p-0.05). CONCLUSIONS: The pilot-run Urolizer shows high analytical reliability. The innovative analysis device may be especially suited for urologists specializing in urolithiasis treatment. The possibility for urine storage at moderate temperatures without loss of analysis quality further demonstrates the applicability of the BRI method.

Calcium oxalate stone formation risk - a case of disturbed relative concentrations of urinary components.

BACKGROUND: Although afflicted with stone formation, urolithiasis patients often present with normal renal excretions of lithogenic and inhibitory substances. In this study, crystal formation is not interpreted as the result of urinary excretions simply exceeding the static limits of normal ranges but rather as the consequence of relative combinations of such parameters which convert urine into becoming potentially lithogenic. Our model embraces different triplet combinations of fundamental urinary risk factors for calcium oxalate (CaOx) crystallization, to characterize different levels of urinary stone formation risk. METHODS: Urinalyses and BONN-Risk-Indices (BRI) were determined for CaOx patients under home conditions, after 1 week of hospitalization, and for healthy controls. The relative urinary concentrations and interdependences of, inter alia, free ionized Ca (Ca2+), bound Ca (Ca b), and oxalic acid (OA) were compared. RESULTS: Three levels of CaOx formation risk can be distinguished: (I): low stone formation risk with an increase in (Ca2+) and concomitant decrease in (Ca b), while (OA) remains almost constant, BRI increases simultaneously; (II) moderate risk at (Ca2+)/(Ca b) approximately 1; and (III) high risk with decrease in (Ca2+)/(Ca b) and simultaneous increase in (OA). CONCLUSIONS: The proposed approach of urinalysis interpretation allows complementary strategy of identification of patterns of disturbed urinary composites leading to calculus formation.

The distribution of crystalline material in obstructed stents--in need for intra-luminal surface modification?

PURPOSE: Usually, hampered urine flow and failing of Seldinger technique leads to the explanation "obstructed ureteral stent" with no further clarification where exactly the obstructions are located. If stent obstruction is caused by intra-luminal biofilm and/or crystal deposits, the need of biofilm reducing coatings on the stent's inside has to be discussed. MATERIALS AND METHODS: We investigated 59 stents from patients in whom acute hydronephrosis and/or acute pyelonephritis required stent replacement and/or usage of Seldinger technique failed. The stents were investigated by X-ray and, after longitudinal cutting, by light-microscopy for occurrence of obstructing material. RESULTS: The inside of 25% of the samples was lined with a thick film composed of blood clots and tiny non-aggregated crystals. Only in these samples X-ray investigation showed a positive result for massive inner encrustations, which in fact may be responsible for stent occlusion. 48% of the stents contained few small domains composed of blood clots and crystals. 27% of the stent samples showed no alteration. CONCLUSIONS: 75% of the "obstructed" stents showed no significant inner deposits. Obstruction of urine transport and failure of Seldinger technique occurred due to other reasons. Thus, coating of the stent's inner surface may be overrated.

Diamond-like carbon coatings on ureteral stents--a new strategy for decreasing the formation of crystalline bacterial biofilms?

PURPOSE: Any catheter material placed in the urinary tract provides a surface for bacterial colonization and, therefore, it is susceptible to encrustation with crystalline bacterial biofilm. Encrustation and blockage by biofilms remain a major complication in patient care. Most patients with indwelling ureteral stents experience irritative symptoms related to these implants and many experience discomfort. MATERIALS AND METHODS: Plasma deposited diamond-like amorphous carbon coatings are well-known for their excellent biocompatibility. A low temperature, low pressure plasma enhanced chemical vapor deposition technology was developed especially for coating polymeric medical implants with diamond-like carbon. We investigated the ability of diamond-like carbon to decrease the formation of crystalline bacterial biofilm as well as stent related side effects and discomfort. Diamond-like carbon coated ureteral Double-J stents were tested in vivo. RESULTS: In 10 patients with heavy encrustation, different underlying diseases and a stent removal frequency of less than 6 weeks due to encrustation a total of 26 diamond-like carbon coated stents were successfully tested for their ability to decrease the extent of crystalline biofilm formation. There was a 2,467-day period of experience with diamond-like carbon coated stents. No primarily stent related complications occurred. No crystalline biofilm formation was observed in vivo. Excellent and facile handling, a less painful replacement procedure and high tolerance of application were reported by physicians and patients. Due to low friction the coated stents could be placed and removed much more easily than standard stents. The frequency and severity of symptomatic urinary tract infections were distinctly decreased. CONCLUSIONS: Diamond-like carbon coating is a new strategy to improve the surface properties of ureteral stents. This novel surface effectively decreases friction, encrustation tendencies and biofilm formation.

Computation and modeling of the stone-growth related urinary depletion effect using "depletion V1.0".

PURPOSE: During metabolic evaluation stone patients often present with calculi in the urinary tract or are suffering from high frequent formation of small microliths passing routinely the urinary tract. These concrements are presumably in a state of continuous growth. Consequently, the concentrations of the lithogenic components in the voided urine must decrease. Thus, treatment schemes inevitably fail to focus on the true pathogenic urinary composition. Instead, they focus on underestimated concentrations. This can attain high clinical relevance. In recent publications, we introduced a complex physical approach and provided mathematical equations which can be solved analytically. However, to date, solving the equations with a pocket calculator remains cumbersome. MATERIALS AND METHODS: Depletion V1.0 was developed to integrate the calculation of the depletion effect into the daily treatment of stone patients. Minimum requirement for Depletion V1.0 is the Java 1.2 platform runtime environment, which is supported on nearly all operation systems including Linux, MacOS X, and Windows. The program can be used directly within a Java-compliant web browser (e.g. Firefox, Mozilla, Internet Explorer) or from the program's storage location. The implemented data base provides stone type relevant parameters. Data input is performed via an easy-to-handle graphical user interface. RESULTS: Results are given as values and interactive plots; computation and update of plots are performed in real time. Result sheets can be exported to platform-independent EPS-format or printed out directly. CONCLUSIONS: Depletion V1.0 enables the medical practitioner to obtain an improved interpretation of the stone patient's health status. As input, only one set of easy-to-achieve clinical standard parameters is required. The program will be available from the authors as freeware.

Why does the Bonn Risk Index discriminate between calcium oxalate stone formers and healthy controls?

PURPOSE: The BRI has been shown to discriminate between calcium oxalate stone formers and controls. BRI is the ratio of the concentration of ionized calcium and the amount of oxalate that must be added to 200 ml urine to initiate crystallization. Higher BRI values are predictive of being a stone former and a value of 1.0 has been found to be the cutoff value to distinguish stone formers and controls. It is not easy to present a consistent argument based on the thermodynamics of calcium oxalate crystallization to account for the success of this index. For instance, why should 2 samples sharing the same BRI but with different ionized calcium and oxalate values have the same likelihood of being obtained from a stone former? MATERIALS AND METHODS: Using data on 195 samples the distribution and interrelationships of measured variables were examined. They were used to calculate illustrative data with which it was possible to examine the effects of varying the parameters and their relationships. RESULTS: Data simulations identified 3 necessary and sufficient conditions that must be met for BRI to be an effective discriminator between stone former and nonstone former urine samples. CONCLUSIONS: The success of BRI can be explained as the natural outcome of there being significantly different distributions (stone formers vs nonstone formers) of the concentration of ionized calcium and the formation product minus activity product difference as well as the correlation between these 2 variables.

The use of risk indices: do they predict recurrence? Yes, they (at least some) do.

A suitable and advisedly used risk index is an effective tool for improving prevention, therapy monitoring and classification of almost unmanageable amounts of analysis data and diagnoses. In contrast to statistically founded indices, causality-based risk indices can provide a fundamental insight into the mechanisms of the underlying pathology. However, understanding of stone formation as the result of many linked and often non-linear individual processes must be further improved. Only in this way can risk indices be optimized or better ones be developed. We are confident that, with consistent research efforts, science will be able to predict recurrence of stone formation more accurately within the next couple of years.

Hyperoxaluria, hypocitraturia, hypomagnesiuria, and lack of intestinal colonization by Oxalobacter formigenes in a cervical spinal cord injury patient with suprapubic cystostomy, short bowel, and nephrolithiasis.

Although urolithiasis is common in spinal cord injury patients, it is presumed that the predisposing factors for urinary stones in spinal cord injury patients are immobilization-induced hypercalciuria in the initial period after spinal injury and, in later stages, urine infection by urease-producing micro-organisms, e.g., Proteus sp., which cause struvite stones. We describe a patient who sustained C-7 complete tetraplegia in a road traffic accident in 1970, when he was 16 years old. Left ureterolithotomy was performed in 1971 followed by left nephrectomy in 1972. Probably due to adhesions, this patient developed volvulus of the intestine in 1974. As he had complete tetraplegia, he did not feel pain in the abdomen and there was a delay in the diagnosis of volvulus, which led to ischemia of a large segment of the small bowel. All but 1 ft of jejunum and 1 ft of ileum were resected leaving the large bowel intact. In 1998, suprapubic cystostomy was performed. In 2004, this patient developed calculus in the solitary right kidney. Complete stone clearance was achieved by extracorporeal shock wave lithotripsy. Stone analysis: calcium oxalate 60% and calcium phosphate 40%. Metabolic evaluation revealed hyperoxaluria, hypocitraturia, and hypomagnesiuria. Since this patient had hyperoxaluria, the stool was tested for Oxalobacter formigenes, a specific oxalate-degrading, anerobic bacterium inhabiting the gastrointestinal tracts of humans; absence of this bacterium appears to be a risk factor for development of hyperoxaluria and, subsequently, calcium oxalate kidney stone disease. DNA from the stool was extracted using the QIAamp DNA stool Mini Kit (Qiagen, Chatsworth, CA). The genomic DNA was amplified by polymerase chain reaction using specific primers for oxc gene (developed by Sidhu and associates). The stool sample tested negative for O. formigenes. The patient was prescribed potassium citrate mixture; he was advised to avoid oxalate-rich food, maintain recommended levels of calcium in his diet, and take live bio-yogurt. Two months later, 24-h urinary oxalate decreased from 0.618 to 0.411 mmol/day; 24-h urine citrate increased from 0.58 to 1.10 mmol/day. Six months later, an oxalate absorption test was performed. The patient swallowed a capsule, soluble in gastric juice, containing 50 mg (0.37 mmol) sodium [13C2]oxalate corresponding to 33.8 mg of [13C2]oxalic acid. The amount of labeled oxalate, excreted in urine, was measured by a gas chromatographic-mass spectrometric assay. Oxalate absorption, expressed as the percentage of the labeled dose recovered in the 24-h urine after dosing, was 8.3% (reference range: 2.3-17.5%). In addition to other conventional measures, oral administration of O. formigenes or lactic acid bacteria mixture to promote bacterial degradation of oxalate in the gut, and thus combat hyperoxaluria, may play a role in prevention of calcium oxalate kidney stones.

Oxalate degrading bacteria: new treatment option for patients with primary and secondary hyperoxaluria?

Current treatment options in patients with primary and secondary hyperoxaluria are limited and do not always lead to sufficient reduction in urinary oxalate excretion. Intestinal oxalate degrading bacteria are capable of degrading oxalate to CO(2) and formate, the latter being further metabolized and excreted via the feces. It is speculated, that both endogenously produced, as well as dietary oxalate can be significantly removed via the intestinal tract. Oxalobacter formigenes, an obligate anaerobic microbe normally found in the intestinal tract has one oxalate degrading enzyme, oxalyl-CoA decarboxylase, which is also found in Bifidobacterium lactis. Other bacteria with possible oxalate degrading potency are lactic acid bacteria, as well as Enterococcus faecalis and Eubacterium lentum. However, specific therapeutic studies on humans are scarce and, except for Oxalobacter, data are not congruent. We found the oral application of Oxalobacter successful in patients with primary hyperoxaluria. However, long-term post-treatment follow-up of 1-2 years showed that constant intestinal colonization is not achieved in most patients. In one patient with constant colonization, urinary oxalate excretion normalized over time. Short-term studies with other bacteria such as lactic acid bacteria did not show a specific reduction in urinary oxalate excretion. O. formigenes might be a promising new therapeutic tool in patients with primary and secondary hyperoxaluria.