Lincomycin treatment of guinea pigs causes formation of pigmented phosphate containing gallbladder sludge and stones.

We studied the mechanism of gallbladder sludge formation in guinea pigs (n = 30) treated with lincomycin (80 mg/kg/day) for 7 consecutive days. At sacrifice (day 8) gallbladders of treated animals contained turbid bile, sludge and in one animal a single gallstone. The precipitates were amorphous on X-ray diffraction. Infra-red spectroscopy revealed calcium phosphate as the major component. Compared to saline-treated controls (n = 15) concentrations of total protein, total phosphate and total bilirubin in gallbladder bile were significantly increased (P less than 0.05). The increase in total phosphate was due to the inorganic component, since phospholipid phosphorus was unchanged. The relative amounts of unconjugated bilirubin and of bilirubin mono- and diconjugates in gallbladder bile were unaffected by treatment as was beta-glucuronidase activity. However, sludge was enriched in unconjugated bilirubin compared to gallbladder bile. This was most probably caused by alkaline hydrolysis of bilirubin monoconjugates. To some extent, disproportionation of bilirubin monoconjugates in bile or sludge, either in vivo or during sample preparation, might also have led to increased unconjugated pigment.

Separation and identification of urinary proteins and stone-matrix proteins by mini-slab sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Mini-slab electrophoresis on sodium dodecyl sulfate/polyacrylamide gel can be used to detect protein in unconcentrated urines containing more than 100 mg of protein per liter. Several urinary proteins--including albumin, transferrin, and the Tamm-Horsfall mucoprotein--precipitate, partly or totally, if urine samples are stored at 4 degrees C for one or two days. The present procedure permits identification of the albumin and transferrin in unconcentrated urines, thus avoiding their partial loss. Stone-matrix proteins in 10 mg or more of urinary calculi also could be detected. Albumin, transferrin, and Tamm-Horsfall mucoprotein accounted for greater than 40% of the total matrix protein in most of the urinary stones analyzed. The precipitation properties of these proteins and their high concentrations in stone matrix relative to those in urine suggest that they may play a role in stone formation and growth.

Reduction of infection stones in rats by combined antibiotic and phosphocitrate therapy.

The potential of phosphocitrate to inhibit infection stones in rats when combined with an antibiotic was studied. A significant reduction occurred in both the number and weight of recovered stones from rats receiving combined treatment with amoxycillin (50 mg./kg. body wt./day and phosphocitrate (112 mumol./kg. body wt./day) for four weeks. The inhibitory responses were attributed to the intact phosphocitrate molecule as administration of citrate in equimolar concentrations did not mimic the observed effects of phosphocitrate. In comparison with non-infected controls, antibiotic treatment alone failed to eliminate total stone growth. However, composition of the stone reverted from predominantly struvite to a mixture of struvite and newberyite as urinary parameters normalized. The studies highlight the usefulness of phosphocitrate to restrict magnesium salt deposition in vivo.

The influence of urinary macromolecules on calcium oxalate monohydrate crystal growth.

The Constant Composition (CC) kinetics method has been used for studying the mineralization of calcium oxalate monohydrate (COM) at sustained supersaturations in the presence of pre-bladder urine and macromolecules isolated from normal urine and kidney and bladder stones. The method is especially sensitive for investigating the inhibitory activities of these urinary macromolecular components (UMMC) and matrix macromolecular components (MMMC) with a coefficient of variation in growth rate of approximately 2%. Significant COM mineral inhibition was observed in a wide molecular weight region of urine components. Urine removed directly from the kidney showed appreciable inhibitory activity towards COM crystallization. Normal urinary proteins and the dissolved precipitate resulting from urine centrifugation were fractionated by gel filtration. The resulting solutions were mostly COM mineralization inhibitors. Electrodialysis was utilized to isolate the MMMC (greater than 7000 d) of renal and bladder calculi. While these solutions inhibited COM crystallization, they were also found to be calcium binders as measured by the calcium electrode.

Crystal-matrix relationships in experimentally induced urinary calcium oxalate monohydrate crystals, an ultrastructural study.

Calcium oxalate monohydrate crystalluria was experimentally induced in male rats by administration of ethylene glycol. The crystalluria particles were separated by filtration and studied by scanning and transmission electron microscopy. They were associated with cellular degradation products. Organic material was present both on the surfaces of crystalluria particles as well as within them and was organized in radial striations and concentric laminations. It is proposed that crystal nuclei were formed by heterogeneous nucleation. These nuclei then adsorbed organic material and aggregated. The organic material polymerized resulting in the formation of a crystal-matrix unit which then grew and incorporated more organic material during the crystal growth.

gamma-Carboxyglutamic acid, a component in human pediatric bladder stones containing calcium salts.

The amino acid gamma-carboxyglutamic acid (Gla) has been previously detected in the vitamin K-requiring blood clotting factors, proteins of calcified vertebrate tissue, renal tissue, plasma protein C, ectopic calcifications, and calcium-containing renal calculi. This paper reports the presence of Gla in the EDTA-soluble, non-dialyzable material recovered from human pediatric bladder stones containing calcium salts. In bladder stones composed of calcium oxalate, uric acid and ammonium acid urate, 73 Gla residues per 1,000 amino acid residues were detected. Bladder stones composed of calcium oxalate, uric acid, ammonium acid urate, and hydroxyapatite contained 48 Gla residues per 1,000 amino acid residues present. No Gla was detected in the predominantly magnesium ammonium phosphate (struvite) bladder stones. These results with human bladder stones from children under 10 years of age are consistent with the observations from adult patients in which Gla was detected in the calcium-containing renal calculi but not in the non-calcium-containing renal calculi. The present study adds to the growing body of information concerning the possible role of Gla in normal and abnormal calcium metabolism.

[Identification of a metabolite of floctafenine in urinary calculi]. / Identification d'un métabolite de la floctafénine dans un calcul urinaire.

There are three 4-amino-quinolines used for their analgesic properties: glafenine, antrafenine and floctafenin. Urinary calculi due to glafenine have been described since 1980. Recently two cases of renal calculi containing antrafenic acid have been reported. We discovered a metabolite of floctafenin in a bladder calculus and describe its identification by infrared spectrophotometry and thin-layer chromatography.