Experimental nephrolithiasis in rats: the effect of ethylene glycol and vitamin D3 on the induction of renal calcium oxalate crystals

Using ethylene glycol (EG) and vitamin D3 as crystal-inducing diet (CID) in rats, we investigated the effect of the dosage of EG on the generation of chronic calcium oxalate (CaOx) nephrolithiasis. We collected weekly 24 hour urines and measured herein the amount of oxalate, calcium, glycosaminoglycans (GAG's), creatinine, protein, alkaline phosphatase (AP), gamma-glutamyl transpeptidase (gamma-GT), and N-acetyl-beta-glucosaminidase (NAG). The potential of these urines to inhibit crystal growth and agglomeration was also evaluated. After four weeks, the kidneys were screened by histology and radiography for the presence of CaOx crystals and the amount of kidney-associated oxalate was biochemically measured. Using 0.5 vol.% EG, only a part of the rats showed CaOx deposition in the renal cortex and/or medulla, without obvious differences between Wistar and Sprague-Dawley (SD) rats. If a dietary EG concentration of 0.75, 1.0, or 1.5 vol.% was used, the amount of kidney-associated oxalate was proportionally higher and CaOx crystal formation was consistently found in all rats. Most crystals were encountered in the cortex, whereas in the medulla and the papillary region, crystals were only occasionally detected. From these data, we conclude that in the chronic rat model, based on EG and vitamin D3, a consistent deposition of CaOx crystals is obtained using a EG concentration of at least 0.75%.     

The significance of uric acid in calcium oxalate nephrolithiasis

The findings of serum diagnosis (urea nitrogen, creatinine, uric acid, calcium, phosphorus) in 247 patients with oxalate stone were divided according to sex, and the excretion of uric acid, calcium and phosphorus was estimated. Compared with the control group, the levels of serum uric acid and serum calcium were statistically significantly raised, the mean level not exceeding the normal range. The serum uric acid level was 25% above the normal range in men and 20% above in women, and in the upper range of normal in one third of each, men and women. The excretion of uric acid in women exceeds the normal level in 32% of cases and in 40% of the men, sometimes considerably. The connection between pathological uric acid levels in the serum and urine and the development of oxalate stone, as well as the therapeutic consequences are discussed.     

Pathological and immunocytochemical changes in chronic calcium oxalate nephrolithiasis in the rat

In the present study, we exposed rats to a crystal-inducing diet (CID) consisting of vitamin D3 and 0.5% ethylene glycol (EG), and we investigated histologically the kidney damage induced by the deposition of calcium oxalate (CaOx) crystals. After 28 days, 50% of the animals had renal CaOx crystals, of which 60% also had small papillary stones. Most crystals were present in the cortex. The occurrence of these crystals coincided with morphological and cytochemical changes: glomerular damage, tubular dilatation and necrosis, and an enlargement of the interstitium. The number of epithelial and interstitial cells positive for the proliferating cell nuclear antigen (PCNA) was increased. Tamm-Horsfall protein (THP) was not only demonstrable in the thick ascending limb of the loop of Henle (TAL), but also frequently in glomeruli, in the proximal tubular epithelium, and in the papilla. In the lumen of the tubular system, it was associated with urinary casts. Reflection contrast microscopy (RCM) showed that the crystals were coated with a thin layer of THP. In spite of the high urinary oxalate concentrations, the above described cellular changes were not observed in CID-fed rats without renal crystals. We conclude, therefore, that in the kidney, the retained CaOx crystals rather than the urinary oxalate ions are responsible for the observed morphological and immunocytochemical changes.     

Renal injury mediated calcium oxalate nephrolithiasis: role of lipid peroxidation

1. Intrathecal (i.t.) administration of nociceptin and high doses of morphine induced allodynia in response to innocuous tactile stimuli, and i.t. nociceptin evoked hyperalgesia in response to noxious thermal stimuli in conscious mice. Here we have characterized the nociceptin-induced allodynia and compared it with the morphine-induced allodynia and the nociceptin-evoked hyperalgesia. 2. Nociceptin-induced allodynia was evoked by the first stimulus 5 min after i.t. injection, reached a maximum at 10 min, and continued for a 50 min experimental period. Dose-dependency of the allodynia showed a bell-shaped pattern from 50 pg to 5 ng kg-1, and the maximum effect was observed at 2.5 ng kg-1. 3. Morphine-induced allodynia reached the maximum effect at 15 min and declined progressively until cessation by 40-50 min. The dose-response curve showed a bell-shaped pattern, similar to that induced by nociceptin, with a maximum effect at 0.5 mg kg-1, five orders of magnitude higher than that of nociceptin. 4. The allodynia evoked by nociceptin and morphine were dose-dependently blocked by glycine, D(-)-2-amino-5-phosphonovaleric acid (D-AP5, an N-methyl-D-aspartate (NMDA) receptor antagonist), gamma-D-glutamylaminomethyl sulphonic acid (GAMS, a non-NMDA receptor antagonist) and methylene blue (a soluble guanylate cyclase inhibitor), but were not affected by muscimol (a gamma-aminobutyric acidA (GABAA) receptor agonist) and baclofen (a GABAB receptor agonist). 5. Morphine did not inhibit forskolin-stimulated cyclicAMP formation in cultured cells expressing the nociceptin receptor. 6. Nociceptin-induced hyperalgesia was evoked 10-15 min after i.t. injection. Nociceptin produced a monophasic hyperalgesic action over a wide range of doses from 5 fg to 50 ng kg-1. The nociceptin-induced hyperalgesia was blocked by glycine only among the agents examined. 7. None of the pain responses evoked by nociceptin and morphine were blocked by naloxone. 8. These results demonstrate that, whereas the mechanisms of the nociceptin-induced allodynia and hyperalgesia are evidently distinct, they involve a common neurochemical event beginning with the disinhibition of the inhibitory glycinergic response. Morphine may induce allodynia through a pathway common to nociceptin, but the nociceptin receptor does not mediate the action of high doses of morphine.     

Role of calcium oxalate monohydrate crystal interactions with renal epithelial cells in the pathogenesis of nephrolithiasis: a review

Renal tubular fluid in the distal nephron is supersaturated with calcium and oxalate ions that nucleate to form crystals of calcium oxalate monohydrate (COM), the most common crystal in renal stones. How these nascent crystals are retained in the nephron to form calculi in certain individuals is not known. Recent studies from this laboratory have demonstrated that COM crystals can bind within seconds to the apical surface of renal epithelial cells, suggesting one mechanism whereby crystals could be retained in the tubule. Adherence of crystals to cells along the nephron may be opposed by specific urinary anions such as glycosaminoglycans, uropontin, nephrocalcin, and citrate. In culture, adherent crystals are quickly internalized by renal cells, and reorganization of the cytoskeleton, alterations in gene expression, and initiation of proliferation can ensue. Each of these cellular events appears to be regulated by extracellular factors. Identification of molecules in tubular fluid and on the cell surface that determine whether a crystal-cell interaction results in retention of the crystal or its passage out of the nephron appears critical for understanding the pathogenesis of nephrolithiasis.     

Adhesion and endocytosis of calcium oxalate crystals on renal tubular cells

The present investigation was designed to study interactions between Madin-Darby canine kidney (MDCK) cells and calcium oxalate monohydrate (COM) crystals and to clarify the significance of these crystal-cell interactions in stone pathogenesis. MDCK cells cultured in the presence of COM crystals showed a time-dependent uptake of crystals; this was specific for COM crystals. In the dynamic model system designed to study these phenomena under more physiological conditions, COM crystals adhered to the cell surface and were subsequently internalized. In this endocytotic process, the microvilli of the cell appeared to play an important role. The observation by scanning electron microscopy of complexes consisting of aggregated COM crystals and cell debris led us to speculate that adhesion and endocytosis of crystals might provide the calculus nidus for aggregation and retention of crystals in the renal tubule. Furthermore, glycosaminoglycans and the macromolecular fraction of human urine were shown to have the ability to inhibit the cellular uptake of crystals. Evidence that similar processes may also occur in vivo was obtained using an experimental stone model in rats. Our experiments revealed that most of the COM crystals adhered to the tubular cells and some crystals were endocytosed by the cell. Thus, these crystal-cell interactions might be one of the earliest processes in the formation of kidney stones. Further elucidation of the mechanism and the regulatory factors involved in this process may provide new insight into stone pathogenesis.     

Effect of growth hormone on urine calcium and serum vitamin D metabolites in renal failure

Growth hormone (GH) causes a modest increase in urine calcium excretion in normal adults, but uremic rats given both GH and calcitriol developed hypercalciuria. Ten short prepubertal children with renal insufficiency treated with recombinant human GH (rhGH) had urine calcium to creatinine (Ca/Cr) ratios and serum vitamin D metabolite concentrations monitored prospectively for up to 24 months. Six were also treated with calcitriol and two with other vitamin D preparations. Mean urine Ca/Cr ratios or mean serum concentrations of 1,25-dihydroxy vitamin D, 24,25-dihydroxy vitamin D, and 25-hydroxy vitamin D did not change significantly during treatment with rhGH. The risk for rhGH-induced hypercalciuria is small in children with renal insufficiency, even when treated concomitantly with a vitamin D preparation.     

Retinal abnormalities in experimental vitamin E deficiency

Physiological and biochemical studies have been carried out longitudinally over a period of 12 months in vitamin E deficient and control rats to gain an understanding of the mechanism whereby vitamin E conserves normal retinal function. Electroretinographic studies indicated that the primary effect of vitamin E deficiency was on the photoreceptors. Ultrastructural studies, however, did not show any morphological changes to the photoreceptors which could explain receptor dysfunction. A 30-40% loss of vitamin A (retinol) was found to be associated with vitamin E deficiency. This could be corrected by repletion with vitamin E, but there was no associated improvement in visual function. An irreversible loss of the long-chain polyunsaturated fatty acids from the retina, increased lipid peroxidation and alterations in membrane fluidity were also detected during vitamin E deficiency. We suggest that a deficiency of vitamin E leads to changes in the membrane microenvironment, which could affect photo transduction by either impairing the ability of rhodopsin to undergo conformational changes to the active form, or by disrupting the hyperpolarising and depolarising processes of the photoreceptors.     

Effect of hydrochlorothiazide therapy on the crystallization of calcium oxalate in urine

1 We examined the case notes of 82 psychiatric out-patients (aged 21-84 years) receiving lithium prophylaxis and with steady-state plasma lithium levels. 2 The mean weight-related daily dose of lithium prescribed decreased by about 50% between the third and eight decades. 3 The corresponding steady-state plasma lithium levels showed a less marked tendency to decrease, this only being seen in the seventh and eighth decades. 4 In patients aged 50 years or over the daily lithium dose required to give a plasma level of 1 mmol l-1 (0.50 mmol kg-1) was significantly lower than that (0.65 mmol kg-1) in patients aged under 50 years (P less than 0?5, Student's t-test). In patients aged 70-79 years this dose was 31% lower than in patients under 50 years. However, interindividual variation was great and it was estimated that age only contributed about 14% to the total interpatient variation. 5 Of the 36 patients under 50 years of age, 42% had minor lithium side-effects and 17% were not optimally controlled with lithium. The corresponding figures for the 46 'older' patients were 46% and 28%. 6 Generally the 50% dosage reduction seemed necessary to compensate for an age-related decrease in lithium excretion and to reduce lithium side effects to a level comparable to that acceptable in younger patients.     

Quantitative analysis on the localization of chondroitin sulfate proteoglycan in renal tissues of patients with calcium nephrolithiasis

BACKGROUND: This study was designed to test the hypothesis that the hospital resources utilized in treating pedestrian trauma would be significantly greater than that for automobile occupants. This was based on previous studies that showed that the demographic features and patterns of injury sustained by the pedestrian population were significantly different from that of automobile occupants. METHODS: A hospital-based study was designed utilizing retrospective analysis of a prospective trauma database. All primary retrievals of pedestrians (n=547) and automobile occupants (n=597) involved in accidents in Central Sydney from mid-1990 to mid-1995 were included. The length of hospital stay, use of the intensive care unit (ICU) and visits to the operating theatre (Standard Resource Cost) were compared. RESULTS: The age and injury severity scores were significantly higher for the pedestrian group. The length of stay (days) for the pedestrians (mean, 12 SD 14; median, 7 interquartile range (IQR) 13), was significantly higher (P < 0.0001 ) than that for the automobile occupants (mean, 7 SD 11; median, 2 IQR 6). The ICU utilization (days) for the pedestrians (mean, 1.3 SD 4.0; median, 0) was significantly higher (P < 0.0001) than that of the automobile occupants (mean, 0.6 SD 2.9; median, 0). The average operating theatre utilization per pedestrian (0.65 visits) exceeded that of automobile occupants (0.43) by 50% (P < 0.0001). CONCLUSIONS: The study confirms that the acute care of pedestrian injury utilizes more hospital resources than that of automobile occupants. Resources should be allocated to meet this need both in terms of hospital reimbursement and overall directives in public health policy.     

Studies on structure of calcium oxalate monohydrate renal papillary calculi. Mechanism of formation

A scanning electron microscopy study of the ultrastructure of 18 calcium oxalate monohydrate papillary calculi was performed with the purpose of establishing the main steps in calculus formation. It is concluded that these calculi originate in a "core" located near the central part of the calculus. Significant quantities of organic matter as well as calcium phosphates can be found in the "core" and at the surface of adhesion to the papilla and, in some cases, fibers and calcified tubules can also be found in the contact zone. In no case did this material affect the crystalline structure of the calculi, indicating that its formation follows the calculus genesis. The study of the compact columnar zone revealed that its formation starts in a practically continuous surface formed by organic matter and crystals that surround the core. This layer favors the growth of oriented calcium oxalate monohydrate crystals upon it. Based on these observations, a feasible mechanism of papillary calcium oxalate monohydrate calculus formation is proposed.     

The evaluation of some biochemical parameters in pyridoxine-treated calcium oxalate renal stone formers

20-, 40- and 60-day old chicks were infected with E. tenella (100000 oocysts). The nature of changes in cholesterol, general fat and lecithin of chicks' blood is the same as at the infection with a small dose (5000) of oocysts of E. tenella. Changes in the lipoid components differ only quantitatively: they are greater at the infection with a greater dose of oocysts that at the infection with a small one.     

Effect of enteric oxygen therapy and vitamin E on the secretion and chemical composition of bile in experimental dystrophy

Effects of enteral oxygenotherapy and vitamin E were studied using rats with acute liver dystrophy caused by CCl4. The enteral oxygenotherapy with simultaneous administration of vitamin E were shown to accelerate the restoration of liver function, to improve the bile formation and secretion, to stimulate the synthesis of initial biliary acids as well as to increase the conjugation of the acids with glycine and taurine, to increase the stabilizing properties of bile.     

Effect of dietary linoleic to linolenic acid ratio and vitamin E supplementation on vitamin E status of poults

Two experiments were conducted to determine the effect of dietary linoleic to linolenic acid (LO:LN) ratio and dl-alpha-tocopheryl acetate (TA) supplementation on selected characteristics of the liver and cerebellum and on vitamin E status of turkey poults from hatch through 22 d of age. In Experiment 1, 1-d-old poults were fed diets containing no supplemental TA (0E) or 150 IU TA/kg diet (150E). Poults fed the 150E diet had greater (P < 0.001) concentrations of alpha-tocopherol (TOC) in the liver and plasma than those fed the 0E diet from 7 to 22 d of age. The 150E diet, however, did not completely overcome the decrease in liver and plasma TOC concentrations observed at these ages. The 150E diet had no effect on poult BW, feed efficiency, or on the weight, protein, lipid, or fatty acid concentrations of the liver. Thiobarbituric acid reactive substances assay of liver and hemolysis assay of red blood cells (RBC) showed that the 150E diet decreased the susceptibility of liver and RBC to in vitro peroxidation at 13 and 22 d of age. In Experiment 2, 1-d-old poults were fed the 0E and 150E diets in a complete factorial arrangement with decreasing ratios of LO:LN (10, 5, and 1). Dietary LO:LN ratio had no effect on RBC hemolysis or cerebellum TOC concentration. As the ratio of LO:LN decreased, the arachidonic acid content of liver and cerebellum lipids decreased. Ratios of n-6 to n-3 fatty acids in liver and cerebellum were directly related to dietary LO:LN at 13 and 22 d of age.     

Use of mannitol and propranolol in the treatment of experimental acute renal failure (histomorphological study)

Experimental research was carried out in order to ascertain whether administration of drugs capable of acting on the renal circulation, such as mannitol and propanol, in addition to improving the renal function, also have a protective effect on the histomorphological alterations induced by the acute renal ischemia. On the basis of the results of the research the Authors conclude by asserting that the combined use of mannitol and propanol has a real protective effect in preventing or attenuating lesions of the kidney caused by serious acute renal failure.     

Inhibitors of calcium oxalate crystallization and urolithiasis

In urolithogenic processes both, promoters and deficit of inhibitors, play an important role. The inhibitory action of added inhibitors (magnesium, citrate, pyrophosphate and chondroitin sulphate) was investigated using the urine of 72 patients with calcium urolithiasis. It was concluded that the deficit of inhibitors seems to be an important cause of stone formation in idiopathic oxalocalcic urolithiasis. Nevertheless, when that specific heterogeneous nucleation takes place it becomes an important factor and the inhibitor plays a complementary role in calcium oxalate urolithiasis.     

Prevalence of renal stones in a population-based study with dietary calcium, oxalate, and medication exposures

Little is known about the epidemiology of renal stones, in spite of the relative frequency of this painful condition. This population-based study examined reported renal stone diagnosis in 1,309 women aged 20-92 years to determine whether renal stones are associated with 1) food or water exposures or 2) lower bone mineral density and an increased likelihood of fractures. Results indicated a renal stone prevalence of 3.4%. The average age at diagnosis was 42 years. Renal stone formation was not associated with community of residence, hypertension, bone mineral density, fractures, high-oxalate food consumption, or ascorbic acid from food supplements. Women with renal stones consumed almost 250 mg/day less dietary calcium (p < 0.01) than did women without stones and had a lower energy intake (p < 0.04). The authors' findings do not support the hypothesis that increased dietary calcium is associated with a greater prevalence of renal stones, nor do they identify renal stones as a risk factor for low bone mineral density. Furthermore, lack of other identifiable environmental correlates and the relatively young age at initial diagnosis suggest that genetic components of renal stone formation need further study.