Dirk J. Kok

Intratubular crystallization events.

Can urolithiasis start as an intratubular event? Under severe hyperoxaluric conditions in animal models at least crystal formation can. Recently models have been presented that assess the chances of crystal formation under more normal conditions. These models describe changes in fluid composition as this passes through the nephron, these conditions being simulated in in vitro experiments. It appears that under naturally occurring intratubular conditions calcium-salt crystallization takes place within the time tubular fluid normally spends in the nephron. Precipitation starts with a calcium-phosphate phase under conditions found in the thin limbs. This crystalline phase then (partly) dissolves when collecting duct conditions are used, thereby inducing formation of calcium oxalates. Under these conditions the latter increase in size by way of crystal growth and agglomeration. Large particle formation and cell adhesion can eventually result in particle retention and subsequent stone formation. Viewing urolithiasis as originally an intratubular event has consequences for in vitro experiments and treatments, which are discussed in this paper.

Calcium oxalate stone agglomeration reflects stone-forming activity: citrate inhibition depends on macromolecules larger than 30 kilodalton.

To evaluate the clinical utility of in vitro calcium oxalate monohydrate (COM) crystallization kinetics measurements and to determine the effect of quantitative removal of urinary Tamm-Horsfall glycoprotein on such measurements, we examined 24-hour, room temperature urine collections of patients from our Stone Clinic and of normal subjects from our research laboratories at Ochsner Medical Institutions in New Orleans, LA, and compared their COM kinetic parameters in vitro before and after urine ultrafiltration (30 kd). Data from 53 calcium oxalate stone-forming patients (26% women; mean age, 47 years) who demonstrated radiographic or other evidence of forming at least one stone were compared with data from 22 healthy volunteers (25% women; mean age, 40 years). Hypercalciuria (> 7.5 mm/24 hr), hyperoxaluria (> 0.5 mm/24 hr), and hypocitraturia (< 2.0 mm/24 hr) were present in 38%, 26%, and 26% of the patient population, respectively. Urinary creatinine, urate, calcium, citrate, phosphate, oxalate, pH, volume, total immunoreactive-disaggregated Tamm-Horsfall glycoprotein, and the urines effects on COM solubility, percent crystal growth inhibition, and crystal agglomeration inhibition [tm] were determined. Calcium oxalate monohydrate agglomeration inhibition, [tm], was reduced in stone-forming patients. It decreased with increasing stone frequency, making [tm] a useful tool for measuring the risk of stone recurrence. Urinary Tamm-Horsfall glycoprotein and citrate concentrations were linearly related to COM agglomeration inhibition. Their effects were synergistic. Tamm-Horsfall glycoprotein removal from urine reduced COM agglomeration inhibition dramatically. Alkali therapy increased urinary citrate concentration and increased [tm].(ABSTRACT TRUNCATED AT 250 WORDS)

Clinical implications of physicochemistry of stone formation

Measuring crystallization processes for assessing the risk of stone formation or the effect of treatment on that risk. In summary, crystallization processes are involved in the risk for stone formation at several levels. Several tests are available for assessing if crystalization is disturbed. It is important to first establish for which part of the urinary tract the information is sought and then adapt the experimental conditions to that site [53]. This information helps in visualizing what is happening inside the urinary tract of a SF and what the treatment measures prescribed may do to change that situation.

Physicochemical considerations in the development and prevention of calcium oxalate urolithiasis.

Calcium oxalate urolithiasis is very common in western societies. In recent years significant progress has been made in identifying and quantitating physico-chemical processes involved in calcium oxalate urinary stone formation. The ability of urine to inhibit the agglomeration of calcium oxalate crystals is an important protective mechanism against stone formation. The process of crystal agglomeration is modulated to a large extent by citrate and it is disturbed in patients with hypocitraturia. Dietary factors, in particular high animal protein intake appear to affect adversely the ability of urine to inhibit calcium oxalate crystal agglomeration. Effective prevention of calcium oxalate urolithiasis should aim at restoring the urines ability to inhibit the agglomeration of crystals and more emphasis should be given to dietary measures.

Measurement of urinary flow rate using ultrasound in young boys and infants

PURPOSE We present a technique for measuring urinary flow rates with ultrasound in male infants and children. MATERIALS AND METHODS Urinary flow rate was measured simultaneously by an ultrasound probe placed around the base of the penis and by a funnel with a rotating disk at the bottom in 30 boys with a mean age of 6.7 years (range 4.5 to 10.5), and by ultrasound in 8 infants with a mean age of 10 months (range 1 to 28). Voided volume was measured with a graded cylinder or calculated from the weight change of diapers in infants. Ultrasound and rotating disk maximum flow rates were calculated. The ultrasound signal was calibrated by comparing the collected voided volume to the area under the curve for that void. The volume calculated from the rotating disk flow rate curve was also compared with the collected volume. RESULTS Both methods yielded similar flow curves. However, ultrasound maximum flow rate significantly exceeded rotating disk maximum flow rate (13 +/- 6 ml. per second, range 5 to 22 versus 10 +/- 4 ml. per second, range 4 to 21, t test p <0.001). The underestimation of the flow rate by the rotating disk method may have been due to adherence of urine to the funnel wall. Rotating disk maximum flow rate was lower and voided volume was underestimated by up to 50% (average 15 +/- 2%) in 21 cases. Ultrasound maximum flow rate averaged 6 +/- 3 ml. per second (range 3 to 11.6 [oldest infant]) in the 8 infants. CONCLUSIONS Urinary flow rates can be measured accurately using ultrasound in boys who produce small volumes and/or who are not toilet trained and also in infants. In future studies ultrasound will be applied to subsets of male infants with bladder dysfunction.

Changes in bladder wall blood oxygen saturation in the overactive obstructed bladder.

PURPOSE Several studies suggest that hypoxia of the bladder wall contributes to bladder dysfunction but the exact relation between bladder function and blood oxygen saturation, a surrogate marker for hypoxia, is not known. We determined bladder wall blood oxygen saturation in vivo in an animal model of bladder outlet obstruction to establish the exact relation between blood oxygen saturation and bladder function. MATERIALS AND METHODS In 8 sham operated and 8 urethrally obstructed guinea pigs we measured blood oxygen saturation of the bladder wall by differential path length spectroscopy before surgery and 8 weeks postoperatively. Urodynamic investigations performed during the whole 8-week period provided data on bladder function. RESULTS Before surgery and 8 weeks after sham surgery blood oxygen saturation in the bladder wall was between 88% and 95% during filling. It decreased during voiding and returned to greater than 90% within 30 seconds. Eight weeks after obstruction saturation was significantly lower than in the sham operated group during filling and voiding. The decrease was positively related to bladder pressure during filling and voiding, and was more pronounced when overactivity was present. Local bladder contractions occurred without a measurable increase in bladder pressure but were associated with a decrease in saturation. CONCLUSIONS A normal bladder maintains a high oxygen saturation level during filling. Bladder obstruction compromises this ability, especially when it involves overactivity. Local bladder contractions without a measurable increase in bladder pressure were associated with a decrease in blood saturation.

Metaphylaxis, diet and lifestyle in stone disease

Abstract Objective: The most common urinary stones (calcium salts, uric acid) form due to genetic factors and lifestyle. This review describes why, if and how medication and lifestyle changes can reduce the risk of formation. Methods: Previous reports were reviewed to obtain information on three aspects of urolithiasis, i.e. epidemiology, mechanisms linking lifestyle and urolithiasis and lifestyle intervention for preventing urolithiasis. Results: Epidemiological evidence links the prevalence of urinary stone formation to general lifestyle factors. Detailed analysis has identified individual lifestyle elements that affect the risk of urinary stone formation. Currently there are several concepts that explain the mechanism of stone formation. Urinary markers like calcium, oxalate, phosphate, uric acid and urinary pH are involved in all these concepts. Many studies show that changing (combinations of) specific lifestyle elements has a favourable effect on these urinary markers. Based on this evidence, protocols have been developed that use a combination of these lifestyle changes and medication to prevent stone formation. In well-controlled studies where patients are optimally informed and continuously motivated, these protocols clearly reduce the stone formation rate. In general practice the result is less clear, because the time and tools are insufficient to maintain long-term patient compliance in the use of medication and lifestyle advice. Conclusion: The risk of stone formation can be reduced in general practice when the patient’s compliance is optimised by providing individualised advice, continuous information, and feedback and incorporation of the advice into a regular lifestyle. The use of ‘e-tools’ might enable this without increasing the time required from the physician.

In vivo measurement of bladder wall oxygen saturation using optical spectroscopy

Current diagnosis, follow-up and treatment of patients suffering from bladder dysfunction are mainly symptom-targeted. A recently recognized cause of continuing bladder function loss is a deteriorated bladder microvasculature. Incorporating this aspect into the clinical diagnostic toolbox may improve treatment results. Recent developments in the field of optical spectroscopy now allow for non-invasive measurement of microvascular blood oxygen saturation in living tissue. We have recently reported pre-clinical data that show that this marker can be successfully measured in an animal bladder. In the animal model the marker differentiated bladders with loss of function from those with normal function. In the present paper, we report on the first in vivo measurement of this marker in the human bladder, as proof of principle, in the muscle of bladders with a normal function.

Timelines of the “free-particle” and “fixed-particle” models of stone-formation: theoretical and experimental investigations

Two major theories on renal stone formation will be reviewed, the “free-particle” and “fixed-particle” mechanisms. These theories combine data on intrinsic factors (inborn metabolic errors), extrinsic factors (diet), renal cell responses and the physico-chemistry and biochemistry of urine into mechanisms of stone formation. This paper describes the specific role of time in both mechanisms. The timeline of crystal- and stone formation was deducted from literature data and was measured for two stones using radioisotope decay analysis. The stones of similar size and composition showed, respectively, a timeline of a few years and a development that took decades. In combination with data on stone architecture and patient characteristics these timelines are explained using the free-particle and fixed-particle mechanisms. Consideration of the timeline of stone formation has clinical implications. We conclude that the fixed-particle mechanism can be a slow process where decades pass between the first formation of a precipitate in the renal interstitium and the clinical presentation of the stone. Added to the fact that the mechanism of this initial precipitation is still ill defined, the conditions that started fixed-particle stone formation in an individual patient can be obscure. Blood and urine analysis in such patients does not necessarily reveal the individual’s risk for recurrence as lifestyle may have changed over time. This is in fact what defines the so-called idiopathic stoneformers. For these patients, prevention of outgrowth of previously formed precipitates, papillary plaques, may be more relevant than prevention of new plaque formation. In contrast, a patient who has formed a stone in a relatively short time through the free-particle mechanism is more likely to show abnormal values in blood and urine that explain the starting event of stone formation. In these patients, measurement of such values provides useful information to guide preventive measures.

LLC-PK1 cells as a model system to study proximal tubule transport of water and other compounds relevant for renal stone disease

LLC-PK1 cells were cultured on a permeable support in a two-compartment culture system. Confluent monolayers received an ultrafiltrate-like solution at the apical side and a plasma-like solution at the basolateral side. The distribution of various solutes, including phosphate, calcium, and oxalate over both compartments was measured in time. The transport of water was monitored by alterations in fluid concentrations of radiolabeled inulin. Bicarbonate, glucose, and phosphate were transported rapidly from the apical to basolateral side of the monolayer. Sodium and chloride were reabsorbed without major consequences for the osmolality in the apical and basal fluid. Calcium and potassium were also reabsorbed, but to a smaller extent than sodium. The luminal concentration of oxalate gradually increased to values that were at least three times higher (12.0 ± 0.4 μmol/l) than those in the contraluminal fluid (3.8 ± 0.1 μmol/l). However, since the luminal rise of oxalate completely matched the rise of inulin in the apical fluid this appeared to be the passive consequence of active water reabsorption rather than of net directed oxalate transport. The LLC-PK1 model could prove useful to study the regulation of proximal tubule water transport and its effect on luminal stone salt concentrations under different physiological conditions.