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A model system is described for the induction of renal calcium oxalate crystals with intraperitineal injections of sodium oxalate in rats. Early crystals are formed predominantly in cortical areas. Massive amounts of calcium are associated with this process, as demonstrated by potassium pyroantimonate staining. Actual crystal formation appears to be an involved process associated with calcium, oxalate, and cellular membranes. Although overt stone formation was not observed, we feel that the intimate involvement of membranes during crystal formation may be similar to that found in renal stones.

Ischemia was produced in the inner cortex of the rat kidney by clamping the pedicle (artery, vein, and ureter) and severing collateral connections. After 30 minutes of ischemia, a slight aggregation of membrane-associated particles was observed in the freeze-fractured plasma membranes. The aggregation was progressive after 60 and 120 minutes of ischemia. These changes were reversible after 15 and 240 minutes of reflow of blood following 30 or 60 minutes of ischemia. The changes were irreversible after 120 minutes of ischemia. The cells were vacuolated after ischemic periods of 30 minutes or longer and after 120 minutes of ischemia the tissue was severely damaged and aggregation of membrane-associated particles was evident in the vacuolar membranes. No changes in the tissue or plasma membrane were observed after 5 or 20 minutes of ischemia.

Injuries caused by hydrochloric acid or Triton X 100 application to the rat bladder urothelium, and the effects on calcium oxalate crystal retention, were examined by light and scanning electron microscopy. Damage due to either compound resulted in desquamation of urothelial cells. The crystals appeared to be retained by a fibrillar material, some of which was identified as fibrin. Heparin treatment of injured urothelium was found to prevent crystal retention.

Calcium oxalate stone disease is the most common human urinary stone disease in the Western Hemisphere. To understand different aspects of the disease, calcium oxalate urolithiasis in the rat is used as a model. Spontaneous calcium oxalate urolithiasis is very rare in rats. Thus the disease is experimentally induced and the rats are generally made hyperoxaluric either by administration of excess oxalate, exposure to the toxin ethylene glycol, or various nutritional manipulations. All the experimental models show renal injury associated with crystal deposition. Calcium oxalate crystals are in most cases intraluminal in renal tubules and often attached to the basal lamina of the denuded epithelium. Rat renal papillary tips and fornices appear to be the preferential sites for the deposition of large calcium oxalate calculi. Where urinary supersaturation of calcium oxalate has been studied the crystal forming rat urines are shown to have higher urinary supersaturation of calcium oxalate than their controls. Oxalate metabolism in the rat is nearly identical to that in humans. Thus, in a number of respects, experimental calcium oxalate urolithiasis in the rat is similar to calcium oxalate stone disease in man.

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.

The tridimensional ultrastructure of the inner cortex of rat kidney has been studied by observing freeze-fractured tissue in stereo. The complex structure of the tubules, fenestrated capillaries and glomeruli is more readily observed through the irregular fracturing of the tissue that occurs in this technique. The ultrastructure of the brush border and interdigitating membranes of the proximal tubules, the structure of the fenestrated endothelial membranes of the capillaries and the ribbon-like appearance of the filtration space between the foot processes of the glomeruli were especially well depicted in stereo images of freeze-fractured renal tissue.

Organic material associated with the calcium oxalate crystals in urinary stones and experimentally induced nephrolithiasis was stained with colloidal iron and analysed by energy dispersive x-ray microanalysis using standard techniques. Iron was positively identified in the stained specimens indicating that some of the organic material is an acidic mucosubstance. The results also indicate that some of the organic material of urinary stones may originate in the kidneys.

Bladder tumors of epithelial origin are rare in children. A 7-year-old male presented with intermittent painless hematuria. An ultrasound examination of the bladder revealed a single polypoid mass which proved to be a transitional cell papilloma.

Calcium oxalate crystalluria was induced in laboratory rats by subcutaneous implantation of potassium oxalate containing mini-osmotic pumps in their intercapsular region. Concentrations of major urinary ions were measured and urinary supersaturations of various urinary salts were calculated using a computer programme. The urines of experimental animals that received oxalate had calcium oxalate crystals and higher supersaturations for calcium oxalate compared to their controls. Oxalate levels of the urines of experimental animals were higher than their controls and this increase was proportional to the increase in urinary supersaturation of calcium oxalate. No significant difference was found in the calcium levels of urines from experimental and control animals.

We have compiled a broad overview of the genesis of urinary stones. Central to the theses given are precise definitions of stone as opposed to stone disease. In terms of mechanisms, a kinetic point of view is essential to any formulation relating to stone formation. It is necessary to comprehend the kinetic approach, an understanding of nucleation, growth and aggregation, if one is to appreciate the final product--the stone. Certain fundamental concepts are reviewed which are critical in grasping these phenomena. Not to be forgotten is that the purely physical chemical aspects of stone formation are insufficient to explain all facets of the process. Therefore, interactions between crystals and tissue are explored in suggesting a unifying hypothesis for upper tract stone disease.

From a structured history of 32 current smokers seen in the pulmonary function laboratory of a community hospital, we determined the number of cigarettes they smoked in 24 h. We also asked them to estimate their cigarette butt lengths from a visual model and to collect all cigarette butts over the next 24 h and mail them to us. We counted the butts, individually measured their lengths, and compared these with their previous estimates in order to validate their claims. While in the laboratory, we determined the level of carboxyhemoglobin in the peripheral blood of each patient. Patients tended to estimate the numbers of cigarettes that they smoked in units of five. Light smokers returned more butts and heavy smokers returned fewer butts than the numbers they reported as usually smoking. All patients were able to precisely estimate the average length of their cigarette butts, and they left butts of consistent lengths. There was no association between the numbers of cigarettes smoked and the butt lengths. Carboxyhemoglobin levels were positively associated with the numbers of cigarettes smoked in 24 h and negatively associated with the butt length and the time elapsed since the last cigarette was smoked, but these were not associated with the amount of tar in the cigarettes or with the number of years the person had smoked.

Changes in the number and sizes of membrane-associated particles have been quantitated in the protoplasmic (P) and exoplasmic (E) fracture faces of the outer membrane of nuclei isolated from the inner cortex following renal ischemia and reflow in the rat. No changes were observed in the inner nuclear membrane. After 20-min ischemia, the number of particles in both fracture faces decreased. With reflow, the total number of particles decreased after both 20- and 60-min ischemia. The partition coefficient (Kp = CPF/CEF) increased from 10 to 11 and 17 at 20- and 60-min ischemia then fell below control values to a Kp of 7 after 120 min. After reflow, Kp steadily decreased except after 20-min ischemia followed by 240-min reflow when Kp began to rise. The sizes of particles were predominantly 60 A in the P face of control outer membranes but became larger after ischemia. After 20- and 60-min ischemia with reflow, the size distribution became more normal. The shifts in particle numbers and sizes seem to indicate modifications within the membrane resulting from ischemia.

To evaluate the role of ESWL in vivo for the treatment of human gallstones positioned on the blast path, a canine model was developed to determine the efficacy of stone fragmentation and the subsequent histopathological injury that occurs as a result of this therapeutic technique. Twenty-four 16- to 20-kg mongrel dogs were divided into five groups: I: ESWL without stone, autopsy at 48 hr (N = 6); II: ESWL with stone (mean diameter 16.8 mm, range = 14-19 mm), autopsy at 48 hr (N = 10); III: ESWL without stone, autopsy at 41-46 days (N = 6); IV: ESWL without stone, autopsy immediately after ESWL (N = 1); V: No ESWL or stone, autopsy 2 hr after anesthesia induction (N = 1). A human gallstone (96% cholesterol) was inserted by cholecystotomy (N = 10) in Group II only. All groups (N = 24) had operative placement of a 6.5 Fr accordion catheter into the gallbladder for radiographic visualization. For each blast path treatment, 2000 discharges were delivered at 18-24 kV. Histopathologically, the Group V gallbladder served as a control. Groups I, II, and IV revealed mild subacute injury; dog gallbladders in Group III showed regression of these changes. Total surface area (TSA) of Group II stones increased from a pre-ESWL mean of 6.60 +/- 0.0.84 cm2 to 53.84 +/- 26.8 cm2 post-ESWL (P less than 0.001). Cumulative post-ESWL fragment sizes for particles in less than or equal to 2-, less than or equal to 3-, less than or equal to 5-, less than 10- and greater than or equal to 10-mm categories represented 32.9, 41.6, 49.4, 74.3, and 100% of pretreatment stone weight, respectively. These data indicate that human gallstones can be fractured to a variable degree when treated on the ESWL blast path and that TSA increased significantly. Gallbladder histopathologic changes appear to be reversible by 41-46 days post-ESWL.

The early phases of calcium oxalate crystal formation in rat kidneys after intraperitoneal challenge with sodium oxalate were examined. From the results we conclude that the crystals are formed intraluminally in the proximal tubules of the renal cortex. The size, number, and distribution of calcium oxalate particles depend on the amount of sodium oxalate injected and the time interval after its injection. Tubular necrosis and dilation were evident after administration of higher doses of sodium oxalate. The papillary tip and corticomedullary junction were the preferential sites for crystal localization after longer periods of injections of higher doses of sodium oxalate.

Male Sprague-Dawley rats were challenged with various hyperoxaluric agents including ammonium oxalate, hydroxy-L-proline, and ethylene glycol. All treatments resulted in increased urinary oxalate. Associated with hyperoxaluria was an increase in urinary levels of renal enzymes, gamma-glutamyl transpeptidase, N-acetyl-beta-glucosaminidase, and alkaline phosphatase. Most of the rats did not demonstrate any significant change in urinary levels of beta-galactosidase. There was a highly significant positive correlation between urinary oxalate and N-acetyl-beta-glucosaminidase.

A procedure based on scanning electron microscopic techniques is described for the identification of crystals in urinary sediments and stones. The crystals are identified by their morphology and elemental composition using scanning electron microscopy and x-ray microanalysis. The procedure has a number of advantages over conventional methods. It is easy to use. It is non-destructive so that both the exterior and interior of the same stone can be separately analyzed. It is the only technique in which information about spatial relationships between various crystals in a stone can be obtained easily. Scanning electron microscopic techniques can detect minor components, and analysis of a wide variety of materials ranging from amorphous substances to microcrystals to macroscopic stones is possible.