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We previously reported the importance of osteopontin (OPN) in the formation of urinary calculus. Since OPN protein is present in normal kidneys, we investigated the difference in OPN at the DNA level between normal subjects and urolithiasis patients. There has not been any genetic investigation of OPN in familial urolithiasis previously reported worldwide. To confirm hereditary predisposing factors for urolithiasis, changes in OPN DNA within a family were investigated in relation to the presence or absence of urinary calculus. Leukocyte OPN DNA from two normal subjects and five patients with urinary calculus was investigated by SSCP analysis: OPN DNA nucleotide sequence was determined, based on the result of SSCP analysis. As a result, a mutation of GCC to GCT, encoding amino acid position 250 (Ala-250) was found. To confirm the frequency of mutation at this site, OPN DNA was extracted from peripheral blood in 36 normal subjects (Con group), 25 patients with familial urolithiasis (FSF), and 40 patients with recurrent urinary calculus and who had had two or more previous episodes (RSF). The degree of mutation at Ala-250 was then examined by restriction fragment length polymorphism (RFLP) method. As described above, the nucleotide codon encoding the amino acid sequence position 250, Ala-250, was GCC in two normal subjects. This is the original codon. In five patients with urolithiasis it was GCT, showing a substitution of C with T. On examining the frequency of this mutation, the ratio of normal homozygous GCC was 11/36 in the Con group, 1/25 in FSF and 1/40 in RSF. The ratio of heterozygous GCC/GCT was 16/36 in the Con group, 15/25 in FSF and 26/40 in RSF, and the ratio of homozygous GCT was 9/36 in the Con group, 9/25 in FSF and 13/40 in RSF. Furthermore, the gene frequency of the normal codon GCC was 0.528 in the Con group, 0.3 in FSF and 0.35 in RSF, showing a significantly higher incidence in the Con group (P < 0.05). The gene frequency of mutated GCT was 0.472 in Con group, 0.7 in FSF and 0.65 in RSF, showing a significantly higher incidence in urolithiasis patients (P < 0.05). On investigating the inheritance of Ala-250 in five families in which both parent and offspring demonstrated urolithiasis, the nucleotide substitution in Ala-250 in parents with urolithiasis was inherited by their offspring. In all five families the offspring developed urinary calculus. This study showed that there is no difference in OPN structure between the Con group and urolithiasis patients. However, it was predicted that due to the frequency of normally coded GCC being high in the Con group a difference in the amount of OPN might be caused by a difference in transcription velocity between the two groups. Furthermore, it was suggested that examining the inheritance of Ala-250 within a family is a diagnostic method for identifying the predisposing hereditary factors for urolithiasis patients. [ABSTRACT FROM AUTHOR]

α2HS-Glycoprotein (HS), a crystal surface binding substance extracted from human urine, is considered to be one of the urinary macromolecular inhibitors in urolithiasis. In the present study, reverse transcription–polymerase chain reaction was used to examine HS mRNA expression, and immunohistochemical staining was used to reveal its localization in the human kidney. The inhibitory effects of recombinant human HS and native human HS on calcium oxalate crystal growth were examined in a seed crystal system. HS mRNA was found to be expressed in the human kidney, and it was located in the epithelial cells of distal and proximal renal tubular cells. However, neither recombinant HS nor native HS had an inhibitory effect on crystals in the protein concentration of urine of healthy humans. HS in the urine, therefore, does not seem to be a potent inhibitor in stone formation. [ABSTRACT FROM AUTHOR]

In a previous study we showed that the magnesium-calcium ratio of tap water is negatively correlated with the incidence of calcium-containing urinary stones. In this study we examined the relationship between the incidence of struvite stones, water hardness and the regional geological features on the basis of our previous study and an epidemiological study of urolithiasis performed in Japan. The magnesium-calcium ratio of tap water was found to correlate positively with the incidence of struvite stones. The tap water magnesium-calcium ratio was high in regions of basalt and sedimentary rock and was low in granite and limestone areas. The incidence of struvite stones in the regions of basalt and sedimentary rock was higher than that in the granite and limestone areas. Thus, this study suggested that the incidence of struvite stones is related to the magnesium-calcium ratio of tap water and to the regional geology, as is the case for calcium-containing stones. [ABSTRACT FROM AUTHOR]

Ninety-seven male patients with idiopathic calcium urolithiasis and 17 normal male subjects were studied to evaluate the mechanism of idiopathic hypercalciuria with an oral calcium tolerance test, which has been useful in differentiating hypercalciuria. The changes in parathyroid function, such as parathormone and urinary cyclic AMP, and calcium after calcium load differed between absorptive hypercalciuria and renal hypercalciuria. We have confirmed that the change in serum calcitonin after calcium load was also different in these two hypercalciurias. The increase in serum calcium was sufficient to reduce parathyroid function but serum calcitonin was unchanged after calcium load in the control group, in patients with normocalciuria, and those with renal hypercalciuria. Although serum and urinary calcium were more elevated in absorptive hypercalciuria than in the other three groups, parathyroid function was not significantly reduced after loading in absorptive hypercalciuria. In this group only, the serum calcitonin was significantly elevated after calcium load. It is reasonable to suggest that, in this group, because parathyroid function is usually suppressed by intestinal hyperabsorption of calcium, parathyroid function may not be further suppressed by even calcium load. Possibly the significant stimulation of calcitonin may compensate for the lack of suppression of parathyroid function and maintain normal serum calcium levels in absorptive hypercalciuria. These results suggest that the change in serum calcitonin is also useful to differentiate abnormalities of calcium metabolism in patients with hypercalciuria. [ABSTRACT FROM AUTHOR]

alpha(2)HS-Glycoprotein (HS), a crystal surface binding substance extracted from human urine, is considered to be one of the urinary macromolecular inhibitors in urolithiasis. In the present study, reverse transcription-polymerase chain reaction was used to examine HS mRNA expression, and immunohistochemical staining was used to reveal its localization in the human kidney. The inhibitory effects of recombinant human HS and native human HS on calcium oxalate crystal growth were examined in a seed crystal system. HS mRNA was found to be expressed in the human kidney, and it was located in the epithelial cells of distal and proximal renal tubular cells. However, neither recombinant HS nor native HS had an inhibitory effect on crystals in the protein concentration of urine of healthy humans. HS in the urine, therefore, does not seem to be a potent inhibitor in stone formation.

We recently reported that osteopontin (OPN) and calprotectin (CPT) are present in the matrix of urinary calcium stones, and that OPN mRNA is expressed in the renal distal tubular cells. In the present study, we examined the immunohistochemical distributions of OPN and CPT in urinary stones. The stones used in this study were passed spontaneously from the upper urinary tract. One half of each of the stones was analyzed with an infrared spectrophotometer, and were shown to be comprised of calcium oxalate, calcium phosphate, uric acid and cystine. The other half of each stone was immersed in tetrasodium ethylenediamine-tetraacetate (EDTA) solution. The half-stones were embedded in paraffin and cut into 5-microm sections. The avidin-biotin-peroxidase complex technique was employed. A monoclonal antibody to human milk-derived OPN and a monoclonal antibody to human granulocyte-derived CPT were used as primary antibodies. The immunochemical study using the OPN and CPT antibodies showed positive staining of the matrix of the urinary calcium stones. The stones showed staining in two distinct zones: a core area was stained with randomly aggregated OPN and CPT, and peripheral layers were stained in concentric circles. On the basis of our observations, it is reasonable to presume that OPN and CPT play roles as the matrix in the structure of urinary calcium stones.

Kampo medicine is a traditional Japanese therapeutic system which originated in China and was used to treat various diseases for hundreds of years. Kampo medicine had been also used for the cure and the prevention of urinary calculi for many years, but the effect and the mechanism of this use of kampo medicine are unclear. We examined the inhibitory effect of the kampo medicine takusha on the formation of calcium oxalate renal stones induced by ethylene glycol (EG) and vitamin D3 in rats. We also investigated the effect of takusha on osteopontin (OPN) expression, which we previously identified as an important stone matrix protein. The control group rats were non-treated; the stone group rats were administered EG and vitamin D3, and the takusha group was administered takusha in addition to EG and vitamin D3. The rate of renal stone formation was lower in the takusha group than in the stone group; thus, the OPN expression in the takusha group was smaller than in the stone group. Takusha was effective in preventing oxalate calculi formation and OPN expression in rats. These findings suggest that takusha prevents stone formation including not only calcium oxalate aggregation but also proliferation.

The canine renal distal tubular cell line Madin-Darby canine kidney (MDCK) forms calcium phosphate microliths during a long-term culture in vitro. We identified osteopontin (OPN) and calprotectin (CPT) from a urinary stone matrix. We recently also detected the expression of OPN and CPT in MDCK cells. The relationship between the mechanism of the stone formation and these stone matrix proteins is not yet known. Here, MDCK cells were cultured and inoculated in the subcutis of nude mice. After 4, 8 and 12 weeks, the inoculated tissues were resected, fixed and immunostained with polyclonal anti-human OPN and polyclonal anti-human CPT antibodies. Some serial specimens were stained with von Kossa's procedure. MDCK cells formed some follicular formations in the subcutis of nude mice at least at 12 weeks after transplantation. At 8 weeks after the inoculation, we detected small calcium phosphate stones with MDCK cells trapped in the follicles. The cells forming the stones also expressed both OPN and CPT. The CPT expression sites coincided with the stone formation sites. We confirmed that MDCK cells inoculated in nude mice had stone-forming potential, and we speculate that OPN and CPT play important roles in stone formation by MDCK cells.

It has been reported that osteopontin (OPN) plays an important role during urolithiasis as well as bone formation. Generation of stones in the urinary tract may be associated with osteoporosis and bisphosphonates are potent inhibitors of bone resorption, being used with effect in the management of bone disease. We therefore investigated the relationship between alendronate, a bisphosphonate derivative, and OPN expression in the kidney. Alendronate was administered to rats made hypercalcemic by treatment with parathyroid hormone-related peptide (PTHrP). The renal expression of OPN was then evaluated at both protein and mRNA levels. OPN expression was enhanced in the distal tubular cells of hypercalcemic rats and was decreased by alendronate. The observed inhibition of OPN expression suggests an ability of alendronate and other bisphosphonates to act as inhibitors of stone formation in the urinary tract.

The formation of urinary stones is presumed to be associated with polymorphism of the osteocalcin gene. The most frequently seen polymorphism is the Hind III type located at the promoter region. This polymorphism has been used as a genetic marker in the search for a correlation between urolithiasis and normal subjects. In our study, a normal control group of 105 healthy people and 102 patients with calcium oxalate stones were examined. The polymorphism was seen following polymerase chain reaction-based restriction analysis. The results revealed no significant differences between normal individuals and stone patients (P=0.978), and distribution of the TT homozygote in the control group (42.9%) was similar to that in the patient group (42.2%). Further categorization of the stone patients into normocalciuric and hypercalciuric groups also revealed no statistical differences from controls. We conclude that Hind III polymorphism of the osteocalcin gene is not a suitable genetic marker of urinary stone disease. Further searches for other polymorphisms on this gene correlated with stone disease are suggested. [ABSTRACT FROM AUTHOR]

Crystallization process has a major role in urolithiasis. In the present study, effect of two indigenous plants extracts namely Boerhavia diffusa and Bryophyllum pinnatum extract was determined on the crystallization of calcium oxalate crystals. Effect on the number, size and type of calcium oxalate crystals was observed. Results showed significant activity of both extracts against calcium oxalate crystallization at different concentrations ( P < 0.05). Size of the crystals gradually reduced with the increasing concentration of both extracts. The number of calcium oxalate monohydrate crystals which are injurious to epithelial cells gradually reduced and at the highest concentration of extracts (100 mg/ml) completely disappeared ( P < 0.05). These results confirm that B. diffusa and B. pinnatum extracts have antiurolithic activity and have the ability to reduce crystal size as well as to promote the formation of calcium oxalate dihydrate (COD) crystals rather than monohydrate (COM) crystals. Control of crystal size and formation of COD rather than COM crystals, in combination with the diuretic action of extracts is an important way to control urolithiasis. [ABSTRACT FROM AUTHOR]

Experimental animal model studies suggest that calcium oxalate (CaOx) crystal deposition in the kidneys is associated with the development of oxidative stress, epithelial injury and inflammation. There is increased production of inflammatory molecules including osteopontin (OPN), monocyte chemoattractant protein-1 (MCP-1) and various subunits of inter-alpha-inhibitor such as bikunin. What does the increased production of such molecules suggest? Is it a cause or consequence of crystal deposition? We hypothesized that over-expression and increased production of MCP-1 is a result of the interaction between renal epithelial cells and CaOx crystals after their deposition in the renal tubules. We induced hyperoxaluria in MCP-1 null as well as wild type mice and examined pathological changes in their kidneys and urine. Both wild type and MCP-1 null male mice became hyperoxaluric and demonstrated CaOx crystalluria. Neither of them developed crystal deposits in their kidneys. Both showed some morphological changes in their renal proximal tubules. Significant pathological changes such as cell death and increased urinary excretion of LDH were not seen. Results suggest that at least in mice (1) Increase in oxalate and decrease in citrate excretion can lead to CaOx crystalluria but not CaOx nephrolithiasis; (2) MCP-1 does not play a role in crystal retention within the kidneys; (3) Expression of OPN and MCP-1 is not increased in the kidneys in the absence of crystal deposition; (4) Crystal deposition is necessary for significant pathological changes and movement of monocytes and macrophages into the interstitium. [ABSTRACT FROM AUTHOR]

Tamm-Horsfall protein (THP) is thought to protect against calcium oxalate monohydrate (COM) stone formation by inhibiting COM aggregation. Several studies reported that stone formers produce THP with reduced levels of glycosylation, particularly sialic acid levels, which leads to reduced negative charge. In this study, normal THP was treated with neuraminidase to remove sialic acid residues, confirmed by an isoelectric point shift to higher pH. COM aggregation assays revealed that desialylated THP (ds-THP) promoted COM aggregation, while normal THP inhibited aggregation. The appearance of protein aggregates in solutions at ds-THP concentrations ≥1 μg/mL in 150 mM NaCl correlated with COM aggregation promotion, implying that ds-THP aggregation induced COM aggregation. The aggregation-promoting effect of the ds-THP was independent of pH above its isoelectric point, but was substantially reduced at low ionic strength, where protein aggregation was much reduced. COM aggregation promotion was maximized at a ds-THP to COM mass ratio of ~0.025, which can be explained by a model wherein partial COM surface coverage by ds-THP aggregates promotes crystal aggregation by bridging opposing COM surfaces, whereas higher surface coverage leads to repulsion between adsorbed ds-THP aggregates. Thus, desialylation of THP apparently abrogates a normal defensive action of THP by inducing protein aggregation, and subsequently COM aggregation, a condition that favors kidney stone formation. [ABSTRACT FROM AUTHOR]

n interpretation of previous and recent observation on calcium salt crystallization and calcium stone formation provide the basis for formulation of a hypothetical series of events leading to calcium oxalate (CaOx) stone formation in the urinary tract. The various steps comprise a primary precipitation of calcium phosphate (CaP) at high nephron levels, establishment of large intratubular and/or interstitial (sub-epithelial) aggregates of CaP. These crystal masses subsequently might be dissolved during periods with low urine pH. On the denuded surface of subepithelial or intratubularly trapped CaP, release of calcium ions can result in very high ion-activity products of CaOx, particularly during simultaneous periods with peaks of CaOx supersaturation. Crystals of CaOx may result from nucleation in the macromolecular environment surrounding the apatite crystal phase. In the presence of low pH, low citrate and high ion-strength of urine, formation of large CaOx crystal masses can be accomplished by self-aggregation of Tamm-Horsfall mucoprotein. Following dislodgment of the initially fixed CaOx stone embryo, the further development into to clinically relevant stone is accomplished by CaOx crystal growth and CaOx crystal aggregation of the retained stone material. The latter process is modified by a number of inhibitors and promoters present in urine. The retention of the stone is a consequence of anatomical as well as hydrodynamic factors. [ABSTRACT FROM AUTHOR]

Our aim was to examine the attachment to, and incorporation of intact, highly phosphorylated osteopontin (OPN) into inorganic (i) and urinary (u) calcium oxalate monohydrate (COM) and dihydrate (COD) crystals. uCOM and uCOD crystals were precipitated from ultrafiltered (UF) urine containing human milk OPN (mOPN) labelled with Alexa Fluor 647 fluorescent dye at concentrations of 0.1-5.0 mg/L. iCOM and iCOD crystals were generated in aqueous solutions at concentrations of 0.01-0.5 mg/L. Crystals were demineralised with EDTA and the resulting extracts analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis and western blotting, or examined by fluorescent confocal microscopy and field emission scanning electron microscopy before and after washing to remove proteins bound reversibly to the crystal surfaces. Binding of mOPN to pre-formed iCOM crystals was also studied in phosphate-buffered saline (PBS) and ultrafiltered (UF) urine. mOPN attached to the {100} faces and to the {010} sides of the {100}/{010} edges of iCOM crystals was removed by washing, indicating that it was not incorporated into the mineral bulk. In both PBS and urine, mOPN was attached to the {021} faces of pre-formed iCOM crystals as well as to the {100}/{010} edges, but was concentrated at the intersection points of the {100} and {121} faces at the crystal tips. Attachment in UF urine appeared to be greater than in PBS and stronger at higher calcium concentrations than lower calcium concentrations. In uCOM crystals, the distribution of fluorescence and patterns of erosion after washing suggested attachment of mOPN to the four end faces, followed by interment within the mineral phase. Fluorescence distributions of mOPN associated with both iCOD and uCOD crystals were consistent with uniform binding of the protein to all equivalent {101} faces and concentration along the intersections between them. Persistence of fluorescence after washing indicated that most mOPN was incarcerated within the mineral phase. We concluded that attachment of mOPN to calcium oxalate crystals is face-specific and depends upon the anatomical and genetic source of the protein and whether the crystals are (1) COM or COD; (2) pre-formed or precipitated from solution, and (3) precipitated from urine or aqueous solutions. Our findings emphasise the need for caution when drawing conclusions about possible roles of OPN or other proteins in urolithiasis from experimental data obtained under inorganic conditions. [ABSTRACT FROM AUTHOR]

Intratubular nephrocalcinosis is defined as the histological observation of calcium oxalate and/or calcium phosphate deposits retained within the lumen of the renal tubules. As the tubular epithelium is the primary interaction partner of crystals formed in the tubular fluid, the role of the epithelial cells in nephrocalcinosis has been investigated intensively. This review summarizes our current understanding on how the tubular epithelium mechanistically appears to be involved both in the initiation and in the course of nephrocalcinosis, with emphasis on in vivo observations. [ABSTRACT FROM AUTHOR]

The mechanisms involved in biomineralization are modulated through interactions with organic matrix. In the case of stone formation, the role of the organic macromolecules in the complex urinary environment is not clear, but the presence of mineralogical ‘signatures’ suggests that some aspects of stone formation may result from a non-classical crystallization process that is induced by acidic proteins. An amorphous precursor has been detected in many biologically controlled mineralization reactions, which is thought to be regulated by non-specific interactions between soluble acidic proteins and mineral ions. Using in vitro model systems, we find that a liquid-phase amorphous mineral precursor induced by acidic polypeptides can lead to crystal textures that resemble those found in Randall’s plaque and kidney stones. This polymer-induced liquid-precursor process leads to agglomerates of coalesced mineral spherules, dense-packed spherulites with concentric laminations, mineral coatings and ‘cements’, and collagen-associated mineralization. Through the use of in vitro model systems, the mechanisms involved in the formation of these crystallographic features may be resolved, enhancing our understanding of the potential role(s) that proteins play in stone formation. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the possible effect of a positive family history on the age at the onset of urinary stone disease and the frequency of subsequent symptomatic episodes relating to the disease. Between March 2006 and April 2009, patients with either a newly diagnosed or a previously documented stone disease were included in the study program. They were required to fill in a questionnaire and divided into two groups according to the positive family history of stone disease; group I comprised patients with a family history for urinary calculi and group II those without. Depending on the data obtained from questionnaires, all patients were evaluated in detail with respect to the age at the onset of the stone disease, stone passage and interventions over time, time to first recurrence (time interval between the onset of the disease and the first recurrence), number of total stone episodes and recurrence intervals. 1,595 patients suffering from urolithiasis with the mean age of 41.7 (14–69 years) were evaluated with respect to their past history of the disease. There were 437 patients in group I and 1,158 in group II. There was no statistically significant difference between the mean age value of two groups ( P = 0.09). When both genders in group I were analyzed separately, female patients tended to have higher rate of family history positivity than males. Comparative evaluation of the age at the onset of the disease between the two groups did reveal that stone formation occured at younger ages in patients with positive family history [ P = 0.01 (males), P = 0.01 (females)] and the mean age of onset of the disease was lower in males than females in group I ( P = 0.01). Patients in group I had relatively more stone episodes from the onset of the disease [ P < 0.01 (2–4 episodes), P < 0.01 (≥5 episodes)]. Male patients were associated with higher number of stone episodes ( P = 0.01). Mean time interval between recurrences was noted to be significantly shorter in group I patients when compared with patients in group II [ P < 0.01 (males), P = 0.02 (females)]. In conclusion, our results showed that urinary stone formation may occur at younger ages and that the frequency of symptom episodes may be higher in patients with a positive family history. We believe that the positive family history for urinary stone disease could give us valuable information concerning the onset as well as the severity of the disease. [ABSTRACT FROM AUTHOR]

Matrix stones are radiolucent bodies that present as soft muco-proteinaceous material within the renal collecting system. Following wide-angle X-ray diffraction (XRD) and scanning electron microscopy (SEM), we homogenized a surgically removed matrix stone, extracted and purified protein, and analyzed samples using tandem mass spectrometry for proteomic composition. Resulting spectra were searched using ProteinPilot 2.0, and identified proteins were reported with >95% confidence. Primary XRD mineral analysis was a biological apatite, and SEM revealed fibrous, net-like laminations containing bacterial, cellular, and crystalline material. Of the 33 unique proteins identified, 90% have not been previously reported within matrix stones and over 70% may be considered inflammatory or defensive in nature. Characterization of other matrix stone proteomes, in particular from non-infectious populations, may yield insights into the pathogenesis of this rare stone as well as the mineralogical process that occurs within crystalline calculi. [ABSTRACT FROM AUTHOR]

The objective of the study is to measure the incidence rate of urolithiasis in a nationwide population-based study by a new methodology. A multi-stage stratified “epsem scheme” sampling was designed among imaging centres in 12 ecologic regions (composed of 30 provinces) while the regions were assumed as independent epidemiologic units. Four temporal phases were determined for sampling to represent four seasons. Imaging-proven positive cases for urinary stones were included and questionnaire filled out by interview. 2,955 new cases of urinary stone were included and the overall pooled yearly incidence rate was 136/100,000 (95% CI, 103–168). The whole positive group had a mean age of 41.5 years (SD = 16.3 years) and composed of 57.9% male. The most common season for presentation of disease was autumn. Geographically, the west and north of Iran, and seasonally, autumn have shown the highest incidence rates for the urinary tract stones. To establish the real burden of urinary stone disease, our study adds a little piece of information to the worldwide epidemiology of urolithiasis. [ABSTRACT FROM AUTHOR]

Crystals of calcium phosphate (CaP) added to solutions with a composition corresponding to that at different levels of the collecting duct (CD) and with different pH were rapidly dissolved at pH 5.0, 5.25 and 5.5. Only minor or no dissolution was observed at higher pH levels. Despite this effect, CaP crystals induced nucleation or heterogeneous crystallization of CaOx up to a pH of 6.1, whereas CaP was the type of crystalline material that precipitated at higher pH. Accordingly, small crystal volumes were recorded at pH 5.5 and great volumes at pH 6.7 4 h after the addition of CaP crystals to the solutions. Dialyzed urine appeared to counteract the dissolution of CaP and to reduce the rate of secondary crystallization. The CaP induced crystallization of CaOx was confirmed by a reduction of 14C-labeled oxalate in solution. The APCaOx required for a nucleation or heterogeneous crystallization of CaOx in the presence of CaP was around 1.5 × 10−8 (mol/l)2. For CaP crystal formation on CaP, an APCaP (aCa2+ × aPO4 3−) of approximately 50 × 10−14 (mol/l)2 appeared to be necessary. The CaOx crystals formed were microscopically found in association with the CaP crystalline material and were most frequently of CaOx dihydrate type. Step-wise crystallization experiments comprising supersaturation with CaP ( Step A), supersaturation with CaOx ( Step B) and subsequently acidification ( Step C) showed that CaOx crystal formation occurred when CaP crystals were dissolved and thereby served as a source of calcium. The ensuing formation of CaOx crystals is most likely the result from high local levels of supersaturation with CaOx caused by the increased concentration of calcium. These experimental studies give support to the hypothesis that crystallization of CaOx at lower nephron levels or in caliceal urine might be induced by dissolution of CaP formed at nephron levels above the CD, and that a low pH is prerequisite for the precipitation of CaOx. The observations accordingly provide additional evidence for the important role of calcium phosphate in the crystallization of calcium oxalate, that might occur both at the surface of Randall’s plaques and intratubularly at the papillary tip. [ABSTRACT FROM AUTHOR]

Strong evidence that osteopontin (OPN) is a determinant of urolithiasis has prompted studies comparing the protein’s urinary excretion in healthy subjects and stone formers. However, reported mean urinary values have varied widely, from <1 μg/mL to more than 20 times that value. Since OPN binds to CaOx crystals, the presence of crystals in urine may cause underestimation of its urinary levels. Using a commercial ELISA, we measured urinary OPN levels in the presence of endogenous or exogenous CaOx monohydrate (COM) and dihydrate (COD) crystals. OPN concentrations decreased in the presence of endogenous and exogenous CaOx crystals, but never below 2 μg/mL. Increasing the urinary calcium concentration decreased detectable OPN levels, possibly as a result of changes in the three-dimensional conformation of the protein. Because calcium concentration and the formation of CaOx crystals cannot be controlled in urine, the use of urinary OPN levels as a biomarker for any human pathology must be seriously questioned, but particularly for the investigation of stone formers in whom hypercalciuria and crystalluria are more common than in healthy subjects. [ABSTRACT FROM AUTHOR]

Urinary glycoproteins play an important role in the modulation of calcium oxalate crystallisation. In several cases, this has been attributed to glycosylation of the proteins as evidenced by urinary prothrombin fragment 1 where there is a correlation between sialylation and calcium oxalate kidney stone disease. In the present study, plasma-derived prothrombin fragment 1 (PTF1) was enzymatically modified in order to generate its asialo and aglyco forms. The parent glycoprotein and its two glycoforms were used in calcium oxalate crystallisation studies to assess the role of the carbohydrate moeity in PTF1’s potent inhibitory activity. The glycans inhibited crystal aggregation and promoted crystal nucleation, but had no effect on crystal growth. The terminal sialic acid residues had a small effect on inhibition of crystal aggregation whereas they contributed significantly to promotion of nucleation. These results indicate that glycosylation of PTF1 governs calcium oxalate crystal nucleation and aggregation but it does not affect the protein’s role in inhibiting crystal growth. Since promotion of nucleation and inhibition of aggregation are both regarded as protective mechanisms against calcium oxalate urinary stone formation, the kringle domain on which the glycans are located is implicated in PTF1’s inhibitory activity. It is speculated that modifications in the glycosylation of urinary PTF1 in stone-formers may regulate its capacity to protect against calcium urolithiasis. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the prevalence, incidence, and risk factors of adult urolithiasis in Iran. A total of 8,413 persons aged over 14 years enrolled in this cross-sectional study. They were questioned on the occurrence of urinary stones during their lifetime (prevalence) and on acute urolithiasis in 2005 (incidence) by 62 general practitioners. The subjects were randomly identified from 30 counties of Iran. Data on risk factors for urolithiasis including age, race, education, body mass index, hypertension, and current use of medication were also obtained by self-administered questionnaire. Of the 7,649 participants who provided information, 5.7% (436) [95% confidence interval (CI) 4.2–5.4], reported urinary stones. The prevalence increased from 0.9% in adults aged 15–29 years to 8.2% in those aged 60–69 years (test for trend, P = 0.001). Urolithiasis was slightly more frequent and persisted in males (6.1%) than females (5.3%) giving a male-to-female ratio of 1.15:1 [odds ratio (OR) 1.03; 95% CI 0.64–1.36; P = 0.814]. The annual incidence of urolithiasis in 2005 was 145.1. The average cumulative recurrence rate was 16% after 1 year, 32% after 5 years, and 53% after 10 years. Urinary stones were more in number among men and women who lived in south central and southwest counties, with odds increasing from west to east and from north to south. A positive association was found between urolithiasis and obesity (OR 1.74; 95% CI 1.21–2.31; P = 0.04), diuretic use (OR 1.62; 95% CI 1.18–2.70; P = 0.03), hypertension (OR 1.88; 95% CI 1.26–2.18; P = 0.04), unemployment (OR 2.10; 95% CI 1.43–2.14; P = 0.04), consumption of tea (OR 1.64; 95% CI 1.32–2.62; P = 0.03), consumption of cola (OR 1.49; 95% CI 1.23–2.19; P = 0.02), and meat consumption (OR 1.38; 95% CI 1.29–2.21; P = 0.02). This study provides a quantitative estimate of the prevalence, incidence, and main risk factors for adult urolithiasis in the Iranian population. Further studies are warranted in order to determine the incidence and prevalence of urolithiasis in different ethnic groups. [ABSTRACT FROM AUTHOR]

Prevention of recurrent stone formation will only be possible with careful metabolic evaluation and appropriate management. In this present prospective study, a total of 95 patients with calcium oxalate (CaOx) stone disease were evaluated with respect to the effects of a calcium channel blocking agent (verapamil) therapy on stone-forming risk factors. A total of 95 patients with CaOx urolithiasis were well evaluated for the possible specific effects of verapamil administration on stone-forming risk factors during long-term follow-up. All patients had calcium-containing stones with normal renal morphology and function without any urinary tract infection. The follow-up period ranged from 12 to 36.6 months, with a mean value of 24.4 months. The age of the patients (54 male and 41 female; M/F: 1.31) ranged from 20 to 46 years (mean 34.3 years). On metabolic evaluation all patients had some kind of risk factors and patients were independently randomized into two groups, namely group 1 ( n = 49): patients receiving calcium entry blocker, verapamil hydrochloride (isoptin 240 mg KKH tablets, oral t.i.d.); group 2 ( n = 46): patients receiving no specific therapy (control patients) that were matched for sex and age. Follow-up results (at least 1 year) with respect to the changes in urinary stone-forming risk factors were recorded in both groups. During long-term follow-up patients undergoing no specific therapy did not show a significant change with respect to the urinary levels of stone-forming risk factors when compared with the others receiving verapamil on a regular basis. In the light of our results as well as the literature data, we believe that the pathophysiological mechanisms underlying the effect of verapamil on stone formation (as a result of enhanced crystal deposition) and on the excretion of the urinary stone-forming risk factors have to be well evaluated in further experimental as well as clinical studies. Although the exact mechanism of action is not clear; we may claim that the limitation of internal calcium shift by these agents may also well effect the tubular process related to oxalate handling which ultimately limits its excretion in urine. [ABSTRACT FROM AUTHOR]

In herbal treatment of kidney stones, anti-lithics are used to “dissolve” the stones or aid their passing to guard against further retention. Diuretic action is also needed to increase the amount of fluid going through the kidneys and flush out the deposits. Previous clinical studies have shown that herbal medicines and their concoctions could be used to inhibit calcium oxalate crystallization. However, the pharmacodynamics and in-vitro effects of such medicines have not been established. Five Chinese herbal medicines were selected based on their usefulness in treating stone disease. A 96-well plate oxalate-induced turbidity in artificial urine was used to evaluate the efficacies of the different herbal medicines on calcium oxalate crystallization. The metastable limit was determined and the nucleation rate was derived from 12-min time-course measurement of turbidity at 405 nm. Phase-contrast microscopy was used to visualize the crystals. The results showed that with increasing concentrations of herbal extracts, smaller calcium oxalate crystal sizes were observed. Overall, the five herbal medicinal extracts tested were able to promote nucleation of calcium oxalate crystals while at the same time decreasing the size. This in-vitro crystallization confirms that prophylaxis of renal stones could be achieved by reducing overall supersaturation through promotion of small crystal nucleates and concomitant pharmacological diuretic action of herbal medicines. Clinical studies will provide more definitive conclusions. [ABSTRACT FROM AUTHOR]

We evaluated the possible effects of a calcium entry blocking agent “verapamil” on new stone formation and/or regrowth of residual fragments after shock wave lithotripsy (SWL) during long-term follow-up (>30 months) and compared the results with the success rates of adequate fluid intake. A total of 70 patients treated with SWL were randomly divided into three different groups, in the first two of which the patients received different preventive measures with respect to stone recurrence and/or regrowth. While 25 patients received a calcium channel blocking agent, verapamil hydrochloride, beginning 3 days before SWL and continued 4 weeks after the procedure, an additional 25 patients were put in an enforced fluid intake program and the remaining 20 patients received no specific medication and/or measure apart from close follow-up. Patients were followed regularly with respect to the clearance/regrowth of the residual fragments and that of new stone formation during long-term follow-up (within a mean follow-up of 30.4 months). The overall stone recurrence rate was 14% (10/70). Of the patients who became stone free (12/25, 48%) in group I, only one patient (1/12, 8.3%) showed a new stone formation during long-term follow-up. The figure was 40% (4/10) in group II patients and 55% (5/9) in group III patients receiving no specific medication. Regarding the residual stone fragments (<5 mm) after SWL, again high fluid intake was found to be the most effective on stone regrowth rates (2/13, 15.3%). Patients treated with verapamil also had acceptable regrowth rates (3/15, 20%). Finally, verapamil treatment significantly improved the clearance of residual fragments; while 7 out of 15 patients with residual fragments passed these particles successfully, (46.5%) in this group; these figures were 46% (6/13) and 18% (2/13) in the remaining groups. Residual fragments located in lower calyces demonstrated a poor clearance rate with higher regrowth rates. Verapamil administration was found to be effective enough to limit the regrowth of residual fragments and also to facilitate residual fragment clearance after SWL. Patients receiving this medication seemed to pass the retained fragments easily in a shorter time than the others. [ABSTRACT FROM AUTHOR]

The progressive increase of the social cost for treating urolithiasis could be related to an increased incidence of the disease and/or to an increase of costs for diagnosing and treating renal stones. In the course of the last century, the incidence of renal stones has progressively increased in Europe, North America, and other industrialised countries. This has been explained in terms of changing social conditions and the consequent changes in eating habits. In contrast, renal stones were less frequent than in developing countries of the world but in the last 20 years investigators began to report high incidences of upper urinary stone disease also from some areas of the Third World concurring with the changing of economic and social conditions. Each stone episode involves the costs for emergency visits, diagnostic work up, and medical or surgical treatment. Furthermore, we have to consider the costs of follow-up visits and the costs of testing and drugs for stone prevention. In adjunct of direct costs for diagnosis and treatment, we should also take into account the indirect individual and social cost of workdays lost. Finally, we should estimate the costs of complications and outcomes of treatment with particular attention to the costs of chronic renal failure secondary to stone disease. The strategy of treatment of each stone centre involves different costs for the treatment of each single stone episode. On the other hand the choice of treatment can be driven by National Health Systems and insurance companies by their policy of reimbursement for different procedures. The trends of renal stone incidence will have different impact on health care systems in different countries. In Europe and North America, the peak of incidence has been probably reached but the increase of costs for diagnosing and treating each single stone episode will still increase the social cost for managing stone disease. For this reason the actual objective should be to optimise protocols avoiding redundant or expensive diagnostic procedures or inappropriate treatments. In developing countries, the incidence of stone disease is still increasing and it could reach peaks even higher as a consequence of hot climate in some geographical areas. In those countries the demand for treatment of symptomatic stones could dramatically increase involving a huge financial outlay. [ABSTRACT FROM AUTHOR]

Urolithiasis is a common complication in patients with hypouricemia. Using a microarea x-ray diffractometer and nanoflow liquid chromatography-mass spectrometry (LC-MS) following SDS-polyacrylamide gel electrophoresis (PAGE), recurrent urinary calculi complicating a hypouricemic patient were analyzed. Analysis with the microarea x-ray diffractometer showed that one of the calculi was composed of calcium oxalate monohydrate and hydroxyapatite. The other was found to be formed from calcium oxalate dihydrate. After determination with LC-MS, both were found to contain uromodulin, albumin, osteopontin, protein Z, and defensins. Lysozyme and calgranulin A were also identified in these calculi. Defensins, which were antimicrobial peptides, and lysozyme, a mucopeptide glycohydrolase, were identified as new organic components of urinary stones. The role of these proteins in the process of urolithiasis is of particular interest. [ABSTRACT FROM AUTHOR]

The present study examines the urinary chemical parameters related to urolithiasis in healthy female volunteers during premenopause and menopause, and discusses the role of menopause in stone formation. We investigated 24-h urine parameters associated with urinary stones and focused upon estrus status. Participants comprised 30 healthy women, 15 childless, premenopausal women and 15 menopausal women without a history of urolithiasis. Our results showed that menopausal women have lower citrate and higher calcium excretion, which might enhance calcium stone crystallization. We propose that the estrus status of female patients should be considered when evaluating metabolic abnormalities. [ABSTRACT FROM AUTHOR]

Crystal adherence to the renal epithelium is widely regarded as a probable mechanism of stone formation. Intracrystalline proteins may provide access to the core of urinary crystals and thereby facilitate the dismantling of these crystals after their attachment to and phagocytosis by renal epithelial cells. The present study investigated the role of proteolysis and limited dissolution of urinary calcium oxalate (CaOx) crystals in South Africa’s white and black populations with a view to understanding the remarkably low stone incidence in the black population compared with the whites. CaOx crystals were precipitated from filtered urine or ultrafiltered urine dosed with an intracrystalline protein, urinary prothrombin fragment 1 (UPTF1), isolated from white and black subjects. The crystals were fractured and subjected to proteolysis and/or limited dissolution before examination using field emission scanning electron microscopy (FESEM). FESEM data showed that CaOx crystals from white and black subjects were eroded by treatment with proteases. Cathepsin D caused the most significant crystal erosion, and more noticeable degradation of CaOx monohydrate (COM) crystals compared to CaOx dihydrate (COD). Limited dissolution studies showed the unique ultrastructures and fragmentation processes of COM and COD crystals. COM crystals appeared to be more susceptible to degradation and dissolution than CODs. Since COMs are predominant in blacks, compared with COD crystals from whites, it is speculated that the lower stone rate amongst South African blacks might be attributed partly to their more efficient destruction of retained COM crystals. [ABSTRACT FROM AUTHOR]

The incidence of urolithiasis has increased in the industrialized nations. However, both the pathogenesis and methods for its prevention remain to be clarified. We demonstrate that the antioxidative effect of green tea decreases the formation of calcium oxalate stones, OPN (osteopontin) expression, and apoptosis, and increases SOD (superoxide dismutase) activity in rat kidney tissues. The inhibitory effect of green tea on calcium oxalate urolithiasis is most likely due to its antioxidative effects. Therefore, we examined oxidative stress in vivo applied to Madin-Darby canine kidney (MDCK) cells, to which catechin, an antioxidant, was added. To evaluate the effects of oxidative stress on MDCK cells, we use a hypoxic condition because hypoxia is known to lead to oxidative stress. Confluent cultures of MDCK cells were exposed to (−)epigallocatechin 3 gallate (EGCG) (0, 0.1, 0.5, 5.0 mg/ml) for 2, 4, 8 or 16 h to determine changes in protein secretion and apoptosis. OPN protein expression was observed in MDCK cells of all 16 groups. The levels of expression of OPN protein were the same among all groups. In all groups, SOD protein expression was observed. In the groups exposed to EGCG 0.5, 5.0 mg/ml, SOD staining was more enhanced than in the EGCG 0 and 0.1 mg/ml groups. No deposits were detected in any of the 16 groups. RT-PCR was performed to detect sequences from OPN (979 bp) and SOD (447 bp). Quantitative analyses showed that SOD activity decreased gradually in all groups. Only in the EGCG 0 mg/ml 16 h group were TUNEL-positive cells observed. In the other groups, TUNEL-positive cells were not detected. EGCG used as an antioxidant protects renal tubular cell from cellular injury caused by oxidative stress through SOD protein expression. [ABSTRACT FROM AUTHOR]

Idiopathic renal calcium stone disease often presents with reduced bone mineral content. Investigations using non-invasive methods for the measurement of bone mineral content (single and dual-photon absorptiometry, dual-energy x-ray absorptiometry, quantitative computed tomodensitometry) show a slight decrease in skeletal mineral content of idiopathic renal stone formers (RSFs).The alterations in bone mineral content in RSFs have different explanations: prostaglandin-mediated bone resorption, subtle metabolic acidosis and 1–25 vitamin D disorders. Bone mineral content is worsened by insufficient dietary calcium leading to a negative calcium balance. [ABSTRACT FROM AUTHOR]

Osteoporosis is associated with the pathogenesis of urinary stone formation. Urinary stones are similar to bone diseases such as osteoporosis and bone metabolism in terms of pathogenesis. Bisphosphonates are potent inhibitors of bone resorption, and are used in the management of bone disease. Furthermore, bisphosphonates have a strong affinity for calcium, and a reported inhibitory effect on calcium oxalate crystallization in vitro. Thus, bisphosphonates might also inhibit urinary stone formation. Madin-Darby canine kidney (MDCK) cells form calcium phosphate microliths at the basolateral side in vitro. We investigated the inhibitory effects of new generation bisphosphonates (alendronate and incadronate) on calcium phosphate microlith formation and on the expression of osteopontin, which is an important urinary stone matrix. MDCK cells formed two types of colonies in three-dimensional soft agar culture; dark colonies containing calcium phosphate microliths and clear colonies free from microliths. We applied purified alendronate and incadronate at concentrations of 10-11, 10-9, 10-7 and 10-5 M to MDCK cells cultured in three-dimensional soft agar and investigated the efficiency of colony formation and the dark colony ratio (number of dark colonies relative to the total number of colonies). The administration of 10-9 and 10-7 M alendronate decreased the dark colony ratio compared with controls, whereas incadronate did not significantly alter this colony ratio compared with controls. The expression of osteopontin in cultured cells was inhibited by the 10-7 M alendronate administration. The present findings show that alendronate inhibits calcium stone formation, suggesting that it is effective in the prevention of urolithiasis. [ABSTRACT FROM AUTHOR]

Previous studies have described the inhibitory effects of citrate on calcium oxalate crystallization in place of crystal growth, but the effects of citrate on matrix proteins of stones has not been studied in vivo. To examine the effect of citrate on the matrix, we investigated the effect of citrate on osteopontin (OPN) expression, which we had previously identified as an important stone matrix protein. Control rats were treated with saline while rats of the stone group were treated with ethylene glycol (EG) and vitamin D3, and the citrate groups (low-dose and high-dose groups) were treated with a citrate reagent compound of sodium citrate and potassium citrate, in addition to EG and vitamin D3. The rate of renal stone formation was lower in the citrate groups than in the stone group. This was associated with a low expression of OPN mRNA in citrate-treated rats relative to that in the stone group. Citrate was effective in preventing calcium oxalate stone formation and reduced OPN expression in rats. Our results suggest that citrate prevents renal stone formation by acting against not only the crystal aggregation and growth of calcium oxalate but also OPN expression. [ABSTRACT FROM AUTHOR]

Urinary calcium stones are a pathological substance, and they show similarities to physiological mineralization and other pathological mineralizations. The expression of messenger (m) RNAs of osteopontin (OPN), matrix Gla protein (MGP), osteonectin (ON) and osteocalcin (OC) in bones and teeth has been described. We previously identified OPN as an important stone matrix protein. In addition, the spontaneous calcification of arteries and cartilage in mice lacking MGP was recently reported, a finding which indicates that MGP has a function as an inhibitor of mineralization. Here, we examined the mRNA expressions of OPN, MGP, ON, and OC in the kidneys of stone-forming model rats administered an oxalate precursor, ethylene glycol (EG) for up to 28 days. The Northern blotting showed that the mRNA expressions of OPN and MGP were markedly increased with the administration of EG, but their expression patterns differed. The OPN mRNA expression reached the maximal level at day 7 after the initiation of the EG treatment and showed no significant difference after 14 and 28 days, whereas the MGP mRNA expression rose gradually to day 28. The in situ hybridization demonstrated that the cell type expressing OPN mRNA was different from that expressing MGP. We suggest that OPN acts on calcification and MGP acts on suppression. [ABSTRACT FROM AUTHOR]

Osteopontin (OPN) is one of the most important components in calcium stone matrix, but its role in stone formation is not clear. Since quantitative data regarding the excretion of OPN are necessary to assess its role, we have developed a quantitative enzyme-linked immunosorbent assay (ELISA) for OPN, and measured the urinary OPN concentrations in urolithiasis patients. Forty-seven men with urinary stones composed chiefly of calcium oxalate participated in the study. The controls were 13 normal healthy male volunteers. Urine samples were collected early in the morning and analyzed by a quantitative ELISA employing purified polyclonal antibodies to synthesized OPN aminopolypeptides. The urinary ratio of the concentrations of OPN and creatinine (OPN/Cre) in the urolithiasis patients (0.039 ± 0.029) was significantly lower than that in the control subjects (0.062 ± 0.030) ( P<0.05). Single stone formers ( n = 26; 0.050 ± 0.020) had significantly higher OPN/Cre ratios compared with recurrent stone formers ( n = 21; 0.031 ± 0.021) ( P<0.05). The results show that OPN excretion in urolithiasis patients was lowered, presumably because of the incorporation of OPN by kidney stones. [ABSTRACT FROM AUTHOR]

We previously extracted crystal surface binding substance (CSBS) from human urine and showed that it appeared to constitute a substantial proportion of urinary macromolecular inhibitors of calcium oxalate crystallization. CSBS was isolated from human urine and fractionated by three consecutive chromatography procedures in order to characterize protein inhibitors of calcium oxalate crystallization. Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and NH2-terminal amino acid sequencing revealed that inhibitory fractions eluted from a final, hydroxyapatite column contained prothrombin and osteopontin. Hydroxyapatite column fractions also contained other, unidentified protein inhibitors of calcium oxalate crystallization. CSBS contained also human serum albumin, α1-acid glycoprotein, α2-microglobulin, α2-HS glycoprotein, retinol-binding protein, transferrin, and Tamm-Horsfall protein, but these proteins seemed to play no direct role in inhibitory activity. [ABSTRACT FROM AUTHOR]

The nutrient intake of 69 stone formers (SFs) from three subsets of the local population (urban 22, rural tribal 22 and rural nontribal 25) and 69 age, sex, weight and socioeconomically matched control subjects (NSs) (urban 20, rural tribal 22 and rural nontribal 27) was studied. Simultaneously their timed 24-h urine samples collected over a similar period were analyzed. In general caloric and protein intake was low in all the groups but was strikingly low in the rural subjects. Intake of all nutrients was lowest in the tribal group. Although no difference was observed in diet between NSs and SFs in the same population subjects, SFs had higher urinary excretion of oxalic acid and calcium and lower excretion of citric acid and excreted more saturated urine. Notably magnesium intake was normal in both NSs and SFs, but mean excretion of magnesium was lower than normal in all the groups, suggesting its defective absorption. The influence of dietary intake of protein, carbohydrate, fat, fiber, calcium and oxalic acid on urinary excretion of calcium, oxalic acid, uric acid, inorganic phosphorus, magnesium and citric acid was examined using the chi-square test. No association was observed, thus suggesting that this low nutrient intake did not influence the lithogenic process. Thus, the overall observations suggest: (a) poor nutrition, (b) no effect of diet on urinary stone disease, (c) no difference in the nutrient intake between NSs and SFs and (d) a higher excretion of promoters and a lower excretion of inhibitors in SFs than in NSs. [ABSTRACT FROM AUTHOR]

This study was undertaken to identify proteins which are found associated with calcium oxalate crystals induced in vitro in normal human and rat urine. Crystallization was initiated by adding sodium oxalate individually to each urine sample without centrifugation and filtration. Crystals were collected and analyzed by scanning electron microscopy and X-ray diffraction. Crystal matrix proteins (CMPs) were obtained by demineralization of the crystals with ethylenediaminetetra-acetic acid (EDTA) and analyzed by western blotting technique for immunological identification. Crystals produced in human urine were found to be a mixture of calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD) while those produced in rat urine were exclusively COD. CMPs extracted from crystals in human urine comprised, in addition to prothrombin-related proteins, osteopontin and albumin. However, CMPs extracted from crystals in rat urine contained only osteopontin and albumin. Prothrombin-related proteins were found only in trace amounts. In a separate experiment, rat urine samples were supplemented with COM before inducing crystallization. Similar results were observed showing that CMP contained osteopontin, albumin and trace amounts of prothrombin-related proteins. We conclude that several urinary macromolecules including not only prothrombin-related proteins, but also osteopontin and albumin, become associated with CaOx crystals. The incorporation of these proteins in growing stones is not only due to the presence of γ-carboxyglutamic acid as it was suggested for prothrombin-related proteins, but may be due to other factors such as urinary chemistry, presence of glutamic and aspartic acid residues, and calcium-binding sites. [ABSTRACT FROM AUTHOR]

The interaction between renal epithelial cells and calcium oxalate (CaOx) crystals and/or oxalate ions plays a critical role in the formation of urinary stones. Epithelial cells respond to hyperoxaluria and the presence of CaOx crystals in the kidneys by increased enzymuria and internalization of the crystals. Crystal cell interaction results in movement of crystals from the luminal to the basolateral side between the cells and the basement membrane. Once beneath the epithelium, crystals adhere to the basement membrane and become anchored inside the kidneys. Crystals anchored to basement membrane of the peripheral collecting duct aggregate with other crystals and move through an eroding epithelium to the papillary surface, furnishing an encrustation platform or a nidus for future development of a kidney stone. Thus interaction between renal epithelial cells and CaOx crystals and/or oxalate ions is an essential element in the development of urinary stone disease. [ABSTRACT FROM AUTHOR]

To determine wether high-energy shock waves possess bactericidal potential, ATCC strains of Eschericha coli, Streptococcus faecalis, Pseudomonas aeruginosa and Staphylococcus aureus were suspended in solution at concentrations approximating 10 bacteria per milliliter, placed in polypropylene cryovials, and immersed in the water bath of a Dornier HM3 lithotriptor. Each cryovial was then fluoroscopically guided to the epicenter of the f2 focal point and 2000 shocks at 20 kV applied. Suspensions were then serially diluted and colony counts obtained. The procedure was then repeated with 4000 shocks at 20 kV from the Dornier HM3 and 4000 shocks at intensity level 4 from a Wolff Piezolith 2200 shock wave lithotriptor. Comparison of shock-wave-treated and sham-treated bacterial suspensions revealed no significant difference in bacterial growth according to the colony count technique. We conclude that high-energy shock waves, whether generated by spark gap or piezoelectric array, do not possess significant bactericidal activity. [ABSTRACT FROM AUTHOR]

A simple and rapid technique for the determination of oxalate in urine by ion chromatography has been described. Although there is difficulty in separating the oxalate peak from the sulfate peak on the conductivity chromatogram of unprocessed urine, it is possible to eliminate the sulfate peak by the addition of barium chloride to the urine. Using this technique, the author has estimated the urinary oxalate in 33 urolithiasis patients and in 40 non-urolithiasis patients. The means of 50 urinary oxalate determinations in 33 urolithiasis patients and of 42 urinary oxalate determinations in 40 non-urolithiasis patients were 21.5±11.4 and 19.5±13.0 mg/gCr, respectively. Of the 33 urolithiasis patients, 17 were calcium stone formers and 6 were non-calcium stone formers whose stones had been analyzed by infrared spectrometry, and the mean urinary oxalate values were 19.4±6.9 and 21.3±8.2 mg/gCr, respectively. The urinary oxalate was significantly higher in children under the age of 10 years. [ABSTRACT FROM AUTHOR]