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Background: The combination of infrared spectroscopy and morphological analysis significantly improves the urinary stone analysis. In addition to common urinary stones, it is not unusual to encounter spurious or factitious stones that, if not appropriately identified, can lead to errors in the diagnosis. In this study, we show the importance of Infrared Spectroscopy and the morphological analysis, for determining the presence of drugs crystals or atypical components in the calculi., Methods: 1041 urinary stones were analyzed by morphocostitutional analysis, in addition the rare stones were analyzed by chemical spot test analysis., Results: Among 1041 calculi analyzed, 1018 had a known composition, 23 samples were stones with rare composition or fake urinary stones., Conclusions: Infrared spectroscopy (FT-IR), allows to identify, theoretically, any substance, including drug-containing calculi or calculi with unusual composition and identify false stones. This is mandatory to treat patients affected by urolithiasis with a personalized clinical approach.

The Aim: According to statistical data of a number of the countries, have an urolithic disease from 5% to 15% of the population today. A recurrence arises at 50-85% of patients. The purpose of this work was complex studying of morphology, structure, mineral structure of urolit, identification of regularities of their formation, establishment of interrelation between mineral and organic matter in structure of urolit., Materials: the urinary stones from the urology departments of hospitals and clinics of the Tomsk region (Russia)., Method: s: crystallomorphological (the study of the surface morphology of uroliths binocular and trinocular microscopes); polarizing optical (study of the mineral composition of uroliths on a polarizing microscope); x-ray crystallography (the study of spectral composition on the apparatus DRON-3); electron microscopic (study of the peculiarities of morphology of crystals in an electron microscope)., Results: On features of morphology of a surface four types of urolit are allocated: druzovidny, sferolitovy, combined, korallovidny. Structural kinds of urolit: crystal and granular, dendritovidny, combined, rhythmic - zone. In structure of urolit the following types of rhythms are established: zone, granular, combined. In the urolitakh with rhythmic - the zone structure allocated elements of their building: kernel, layer, zone, rhythm. A mineral part of uric stones is presented by the crystals belonging to the classes: oxalates, phosphates, urat ., Summary: The peculiarities of the morphology of the selected four types of uroliths: druzoid, with, combined, coral. Structural variations uroliths: crystal-grained Shelly, dendritic, combined, rhythmically zoned. In the structure of uroliths, the following types of rhythms: zoned, granular, combined. In urolith with rhythmically zoned structure selected elements of their structure: core, layer, area, rhythm., Discussion: Morphological and structural features of the structure of urolites, especially the presence of rhythmic zoning, due to the alternation of layers of mineral and organic matter, indicate a close relationship between the living organism and the organo-mineral aggregate in the human urinary system. With a high degree of confidence we can talk about the symbiosis of living and mineral matter in the human body; as a result of this symbiosis, organic-mineral formations are formed, which are often the further cause of some diseases (for example, urolithiasis and cholelithiasis)., Conclusion: The complex research of uric stones allows to obtain important information on their structure, structure and features of morphology. Morphological and structural features of the structure of urolit, in particular existence of the rhythmic zonality caused by alternation of layers of mineral and organic matter demonstrate close interrelation between a live organism and the organo-mineral unit in an urinary system of the person.

Urolithiasis is a frequent and in many cases serious disease. Proper analysis of kidney stone composition is crucial for appropriate treatment and prevention of disease recurrence. In this work, scanning electron microscopy (SEM) coupled with energy-dispersive spectroscopy was applied for a study of 30 samples covering the most common types of human kidney stones. The results are analyzed and evaluated in terms of applicability of the method for both routine kidney stone analysis as well as collecting of specific data. The method provides complex information about studied samples including morphology of the stones and of the present crystals or their aggregates. It also brings information on elemental composition of the phases. After application of standardization, quantitative microanalysis with detection limits of 400 ppm (Mg, P, S, Cl, K, Ca), 500 ppm (Na) and 1200 ppm (F) was obtained. Compositional mapping with EDS shows the elemental distribution within a sample. This study demonstrated that information on morphology and chemistry acquired by these methods was highly reliable for identification of phases, even when present in small amounts. It provided information on kidney stone structure, relationships between phases, major and minor element content, and variations in chemical composition related to the growth of the stones. SEM represents a powerful tool in urinary stone analysis, since a single facility can produce a wide spectrum of information. It can be suggested as a basic method used for routine urinary stone identification, whilst bringing additional detailed information that cannot be obtained by other methods.

Urinary stones are complex mineralogical formations in the urinary system often impairing the kidney function. Several studies have attempted to understand the mechanisms of stone formation and growth; however, it remains to be fully explored. Here, we present a detailed investigation on the morphological and mineralogical characterizations of urinary stones. Structural properties of different types of urinary stones were done by X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), and field-emission scanning electron microscope (FE-SEM) analyses. X-ray maps of major and the trace elements were obtained using electron microprobe (EPMA) technique. Basic metabolic panel and urinary parameters of the patients were used for comparing mineral compositions among stone types. The study included five major types of stones identified based on the FTIR spectra. FTIR and XRD helped in identifying the major components of these stones. FE-SEM images revealed distinct microstructure and morphology of the stones among the stone types. EPMA analysis showed the presence of many metals other than calcium and certain non-metals within the urinary stone matrix at measurable levels, sometimes with distinct distribution patterns. The study demonstrates the characteristic micro-structure, morphology, distribution, and composition of elements in different stone types. Findings of the study provide scope for understanding the complex mechanisms involved in the urolithogenesis and association of trace elements in it.

The aim of the work was to present current concept of the pathogenesis of urolithiasis. Treatment and prevention of this disease a challenging issue. The article presents basic information about Randalls plaques that are described as calcium salt deposits on the surface of the transitional cell epithelium. The cause of Randalls plaques was the subject of many studies and is still not completely clear. To date, we can state that the deposit formation starts in the pelvicalyceal system and is directly linked to recurrent urolithiasis. The discovery of Randall plaques in the 1940s transformed the conception of stone formation, but there are even more questions about the pathogenesis of urolithiasis. In that respect, we consider it important to analyze the studies on Randalls plaques.

Aim: To successfully treat and prevent urolithiasis, the composition and structure of uroliths should be examined using modern analytical techniques. For a long time, studies of the biomineral formation in the human body have been conducted exclusively in the field of medicine. The main attention has been paid to the diagnosis and treatment of diseases leading to the occurrence of pathogenic formations. At the same time, it is quite obvious that it is important to have a clear idea about the causes of pathogenic formations, the mechanisms for their further formation, composition and structure. Currently, these issues are widely studied all over the world by mineralogists, biochemists, geo-ecologists using methods of mineralogical analysis. The aim of this work was to determine the content of the elements that make up uroliths. This value should be normalized by the clarks of the noosphere. Studies on the mineral composition and structure of uroliths in the Tomsk region allowed to calculate the content of elements. It turned out that each medical district has its own specific geochemical series, which depends, probably, on natural and technogenic factors., Materials and Methods: The study included urolith samples obtained from residents of 4 medical sub-districts of Tomsk district. 100 samples of different mineral composition were studied. Analytical techniques including crystal-morphological, spectral, X-ray structural, instrumental neutron-activation, electron microscopic analyses were used to investigate the morphology, mineral composition and structure of uroliths., Results: The average content of elements in the uroliths, and the element concentrations normalized by the clark were established. After calculating the concentration, geochemical associative series of elements were formed. Depending on the morphology, drusiform, microspherolite, porous, coral formations, uroliths with a smoothed surface and stones with combined morphology were identified. Three groups of uroliths were defined according to their structural features: crystalline-granular, spherolithic and combined., Conclusion: Studies to determine the mineral composition and structure of the uroliths of the inhabitants of the Tomsk region made it possible to calculate the content of the elements that make up uroliths, normalized by clarks of the noosphere. Each medical sub-district has its own specific geochemical series, which depends on natural and technogenic factors. The increased value of the indicator of the content of elements in uroliths should serve as a warning factor for developing measures to reduce the urolithiasis incidence in the population.

To better understand the basis of urinary calculi formation, we studied the composition, microstructure, and element analysis of different types of urinary calculi. Sixty people with urinary stones in Shanxi province were selected randomly. The composition of urinary stones was analyzed using Fourier transform infrared spectroscopy. The microstructure of material components was observed by a scanning electron microscopy and the elemental distribution and composition were analyzed by an X-ray energy spectrometer. Furthermore, general information, BMI, history of medicine, chronic medical history, family history, and recurrence rates were collected. Female-to-male ratio was 1:2.5; median age was 43.2 years old. Of the patients, 13.3% were found definite family history and 46.6% of patients for recurrence history. It was found that mixed stones account for the largest proportion (65%), followed by calcium oxalate monohydrate calculi (26.67%). In mixed stones, the mixture of calcium oxalate monohydrate and hydroxyapatite had the largest proportion, accounting for 71.79%. Stones showed different microcosmic characteristics and element distribution. Stones varied widely in distribution, infrared spectrum, microstructure, and element composition, which provided an important basis for urinary calculi research regarding urinary stone formation., (© 2016 Wiley Periodicals, Inc.)

Seven human urinary stones were collected from urinary bladders of patients hailing from Istanbul, Turkey. Their XRD, EDX, FT-IR and FT-Raman spectra as well as SEM images have been recorded to determine their chemical compositions, morphologies, crystal structures, and crystallite sizes. XRD and vibrational (FT-IR and FT-Raman) analyses indicate that six out of the seven stones have identical contents. The ratios of organic and inorganic contents of the stones have been determined by their thermogravimetric analyses. The stones have been found to contain calcium oxalate monohydrate and apatite as the major components., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Objective: To compare the accuracy between conventional computed tomography (CT) and dual-energy CT (DECT) in predicting stone composition in a blinded, prospective fashion., Methods: A total of 32 renal stones with known composition were scanned in vitro, first using standard CT techniques at 120 kilovolt peak (kV[p]) and then using fast-switched kilovolt DECT at 80 and 140 kilovolt peak (kV[p]). For the DECT scan, a spectral curve was created demonstrating the change of Hounsfield units (HU) across the kiloelectron volt spectrum. The composition of each stone was estimated by comparing each sample curve with curves of known materials. To attempt stone determination using single-energy CT, the HU of each stone was compared with ranges reported in previous studies. The accuracy of each method was compared., Results: Included were 27 stones large enough to allow analysis. Single-energy measurements accurately identified 14 of 27 stones of all composition (52%), whereas the DECT spectral curves correctly identified 20 (74%). When analyzed by stone type, single-energy vs DECT correctly identified 12 vs 12 of the 12 uric acid stones, 2 vs 3 of the 6 struvite stones, 0 vs 3 of the 5 cystine stones, and 0 vs 2 of the 4 calcium oxalate stones, respectively. When simply attempting to differentiate uric acid vs nonuric acid stones, single-energy CT could accurately differentiate only 6 of 15 stones as nonuric acid (40%) compared with 14 of 15 stones (93%) for DECT., Conclusion: DECT appears to be superior to conventional CT in differentiating stone composition and is particularly accurate in differentiating nonuric acid from uric acid stones., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The term nanobacteria, sometimes referred to as nanobacteria-like particles (NLPs), is presently recognized as a misnomer for inert calcified nanoparticles. However, misinterpretation of its propagation as a living organism still continues. Ultrastructural and elemental analyses, combining immuno-electron microscopy with an original NLP isolate (P-17) derived from urinary stones, and an IgM monoclonal antibody (CL-15) raised against P-17 have now revealed that, oxidized lipids with acidified functional groups were key elements in NLP propagation. Lamellar structures composed of acidic/oxidized lipids provided structural scaffolds for carbonate apatite crystals. During in vitro culture, lipid peroxidation induced by γ-irradiation of FBS was a major cause of accelerated NLP propagation. In pathological tissue samples from hyperlipidemic atherosclerosis-prone mice, CL-15 co-localized with fatty plaques, macrophage infiltrates and osteocalcin staining of aortic valve lesions. These observations indicate that naturally occurring NLP composed of mineralo-oxidized lipids complexes are generated as by-products rather than etiological agents of chronic inflammation., From the Clinical Editor: The term "nanobacteria-like particles (NLPs)" is presently recognized as a misnomer for inert calcified nanoparticles as opposed to living organisms. This study convincingly demonstrates that naturally occurring NLPs composed of mineralo-oxidized lipid complexes are generated as by-products rather than etiological agents of chronic inflammation., (© 2014.)

Synthetic calcium phosphate (CaP) concretions in the form of hemispheres formed under carefully controlled conditions (pH 7.4, 37 °C, no evaporation) from stagnant simulated body fluid on anionic polymeric substrate (laminin) were observed by scanning and transmission electron microscopy and atomic force microscopy. The hemispheres with diameter between approximately 50 and 200 μm were composed of closely connected round spherical and elliptical objects of diameter varying from 70 to 120 nm with surface layer composed of tightly packed spherical objects of diameter 25-30 nm. The phase composition of concretions consisting of amorphous material was uniform. The concretions were formed by aggregation of CaP clusters (Posner's clusters Ca9(PO4)3 or [Ca3(PO4)2] n ) generated in the solution by perikinetic coagulation, their settling onto a substrate and subsequent accumulation through surface migration (surface nucleation) followed by accretion of nanoparticles arriving from surrounding solution. The similarity of ultrafine structures of these synthetic CaP concretions and the compact phosphatic phase appearing in some phosphate calculi indicates that analogous mechanisms could be active at the formation of the latter in the kidneys.

The chemical composition and microstructure of five urolith samples (4 bladder stones and one kidney stone) associated with the feeding of high level of cottonseed meal (CSM) diet to Chinese merino fine wool sheep (Junken breed, Xinjiang) were examined by optical microscope, X-ray diffraction, X-ray energy dispersive spectrometry (EDS), scanning electron microscopy (SEM), and infrared spectroscopy analysis. The bladder stone samples appeared yellow or white, small powder and loose mass, and as finely granular under the optical microscope. However, the kidney stone samples from a experimental sheep were found as small brown mass, higher hardness, and as a cracklike structure. Oxygen, phosphorus, potassium and magnesium were found as four major elements in these uroliths by X-ray energy dispersive spectrometry (EDS). Potassium magnesium phosphate (MgKPO(4)) and potassium magnesium phosphate hexahydrate (MgKPO(4)·6H(2)O) were major components in the bladder stones, while less magnesium ammonium phosphate hexahydrate (MgNH(4)PO(4)·6H(2)O) examined by X-ray diffraction and infrared spectroscopy analysis. However, the newly found prismatic crystals, which were rich in magnesium and pyrophosphate, were identified as magnesium pyrophosphate (Mg(2)P(2)O(7)) in the kidney stone. The bladder stone samples appeared irregular mass and balls, cracked under SEM with low magnification, while appeared cracked, irregular layer-like, honeycomb-like or tiny balls under high magnification. The kidney stone samples were observed as cone, irregular block or layered crystal structures., (Crown Copyright © 2010. Published by Elsevier India Pvt Ltd. All rights reserved.)

Objectives: To report our experimental results on detection and isolation of nanobacteria-like particles (NLP) from urinary stone samples., Methods: From March 2001 to August 2003, 47 urinary stone samples from Japanese patients and 18 from Paraguayan patients were collected and used for compositional analysis, direct survey of NLP by scanning electron microscopy (SEM) and their cultural isolation. For the isolation, culturing was carried out using strict aseptic techniques. Dulbecco's modified Eagle medium with 10% gamma-irradiated fetal bovine serum was used based on the original method described by Kajander and Ciftçioglu., Results: Positive NLP detection rates for Japanese and Paraguayan patient samples were 61.7% (29/47) and 66.7% (12/18), respectively. Positive NLP isolation rates for Japanese patient samples were 20.6% (7/34) and 20.0% (2/10) for Paraguayan patient samples. In the initial isolation, markedly different periods of incubation time were needed for each of the nine cases (6-220 days; median 36 days). Positive detection and isolation were obtained in stone samples with or without calcium phosphate. Growth modes and morphogenesis of NLP were divided into two phases; rod-shaped NLP was detected mainly as a floating form growing in culture medium and spherical NLP with a characteristic apatite shell was detected as an attached form growing on the surface of culture dishes., Conclusions: Lifeless calcifying nanoparticles can be isolated from various human urinary stones, cultured in cell culture mediums and show two characteristic growth phases., (© 2011 The Japanese Urological Association.)

The chemical composition and ultrastructure of urinary calculi obtained from male Boer goats were studied using qualitative chemical analysis, scanning electron microscopy, X-ray diffraction, X-ray energy dispersive spectrometry and Fourier transform infra-red spectroscopy. The calculi came from 10 naturally-occurring cases of urolithiasis and from seven cases of urolithiasis experimentally-induced by feeding a cottonseed meal-rice straw diet supplemented with magnesium oxide. The results indicated that the major component of urinary calculi collected from naturally-occurring and experimentally-induced cases of urolithiasis was struvite (magnesium ammonium phosphate). The study also identified previously unreported prismatic crystals in the uroliths of goats, similar to struvite but rich in potassium. The characteristic ultrastructure of struvite uroliths is described along with a brief discussion of their formation., (Copyright © 2009 Elsevier Ltd. All rights reserved.)

The difference of urine crystallites under 1000 nm in 10 patients with urolithiasis and 10 healthy subjects with no history of urolithiasis was comparatively studied with the nanoparticle size analyzer. By comparing the differences of intensity-autocorrelation curve, polydispersity index (PDI), Zeta potential, and relative error of average diameter of the two kinds of urine crystallites, it was concluded that the urine crystallites of healthy subjects were more stable than those of patients. The urine crystallites of healthy subjects had a narrower size distribution from 100 nm to 350 nm and a better dispersion (PDI < 0.3). However, the urine crystallites of patients with urolithiasis had a wider distribution from dozens of nanometers to 1000 nm and a worse dispersion (PDI > 0.5). The best processing method for urine crystallites detection was found: after antisepticising and protein-coagulating with formaldehyde, the urine was diluted with distilled water of the same volume, then filtrated through a micropore film of 3 microm, and the filtrate was centrifugalized at 4000 rpm for 15 minutes. This method can remove the cell fragments and macromolecular substances in the urine without affecting the detection of the urine crystallites under 1000 nm. The results were consistent with those obtained by transmission electron microscope (TEM).

Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors, which belong to the nuclear receptor superfamily. Some PPARgamma agonists, such as pioglitazone, and dual PPARgamma/PPARalpha agonists, such as muraglitazar, induced urothelial bladder tumors in rats but not in mice. In this study, we investigated the early effects in the urine and bladder of rats treated with pioglitazone to evaluate the possible relation between urinary solids formation and urothelial cytotoxicity and regenerative proliferation. In a 4-week experiment, treatment of rats with 16 mg/kg pioglitazone induced cytotoxicity and necrosis of the urothelial superficial layer, with increased cell proliferation measured by bromodeoxyuridine labeling index and hyperplasia by histology. It also produced alterations in urinary solid formation, especially calcium-containing crystals and calculi. PPARgamma agonists (pioglitazone and troglitazone) in vitro reduced rat urothelial cell proliferation and induced uroplakin synthesis, a specific differentiation marker in urothelial cells. Our data support the hypothesis that the bladder tumors produced in rats by pioglitazone are related to the formation of urinary solids. This strongly supports the previous conclusion in studies with muraglitazar that this is a rat-specific phenomenon and does not pose a urinary bladder cancer risk to humans treated with these agents.

Several modalities of stone analysis are utilised in different laboratories. However, the treating clinician finds it hard to assess the initiation and progression of stone formation. The pathogenesis of calculogenesis still remains a mystery. The purpose of this paper is to assess the pathological mechanisms of stone nucleation and growth by observing the ultra microscopic morphology of the different layers of laminated stones; 130 fragments from 28 randomly selected laminated stones of more than 10-mm diameter were analysed. Wet chemical analysis of the stones was performed. Surface and cross-sectional morphology of the entire stones and the individual fragments was assessed using optical microscopy and images were recorded using ordinary camera. They were further analysed using FTIR for confirmation. By morphological analysis, whewellite, weddellite, uric acid, and phosphate were the main minerals identified. Mixtures of these minerals were also found. Concentric lamination, radial striation, frond formation, and amorphous pattern were the main cross-sectional morphologies obtained. The calculi analysed had differences in their outer and inner portions. This was more pronounced in stones containing predominantly whewellite and uric acid. Whewellite was the outer component in most mixed stones. Uric acid was more in the inner layers of mixed stones than the surface.

Stone analysis is incompletely done in many clinical centers. Identification of the stone component is essential for deciding future prophylaxis. X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM) still remains a distant dream for routine hospital work. It is in this context that optical microscopy is suggested as an alternate procedure. The objective of this article was to assess the utility of an optical microscope which gives magnification of up to 40x and gives clear picture of the surface of the stones. In order to authenticate the morphological analysis of urinary stones, SEM and elemental distribution analysis were performed. A total of 250 urinary stones of different compositions were collected from stone clinic, photographed, observed under an optical microscope, and optical photographs were taken at different angles. Twenty-five representative samples among these were gold sputtered to make them conductive and were fed into the SEM machine. Photographs of the samples were taken at different angles at magnifications up to 4,000. Elemental distribution analysis (EDAX) was done to confirm the composition. The observations of the two studies were compared. The different appearances of the stones under optical illuminated microscopy were mostly standardized appearances, namely bosselations of pure whewellite, spiculations of weddellite, bright yellow colored appearance of uric acid, and dirty white amorphous appearance of phosphates. SEM and EDAX gave clearer pictures and gave added confirmation of the stone composition. From the references thus obtained, it was possible to confirm the composition by studying the optical microscopic pictures. Higher magnification capacity of the SEM and the EDAX patterns are useful to give reference support for performing optical microscopy work. After standardization, routine analysis can be performed with optical microscopy. The advantage of the optical microscope is that, it is easy to use and samples can be analyzed in natural color.

Several plant preparations are administered for treatment of stone disease without scientific basis. This paper presents the results of in vitro and animal experimental studies using scanning electron microscopy (SEM) in the identification of the therapeutic properties of trial drugs in medicine. In the first set of the study, urinary crystals namely calcium oxalate monohydrate and calcium oxalate dehydrate were grown in six sets of Hane's tubes in silica gel medium. Trial drugs namely scoparia dulcis Lynn, musa sapiens and dolicos biflorus were incorporated in the gel medium to identify the dopant effect of the trial drugs on the size and extent of crystal column growth. The changes in morphology of crystals were studied using SEM. In the second set, six male Wistar rats each were calculogenised by administering sodium oxalate and ethylene glycol and diabetised using streptozotocin. The SEM changes of calculogenisation were studied. The rats were administered trial drugs before calculogenisation or after. The kidneys of the rats studied under the scanning electron microscope showed changes in tissue morphology and crystal deposition produced by calculogenisation and alterations produced by addition of trial drugs. The trial drugs produced changes in the pattern of crystal growth and in the crystal morphology of both calcium oxalate monohydrate and calcium oxalate dihydrate grown in vitro. Elemental distribution analysis showed that the crystal purity was not altered by the trial drugs. Scoparia dulcis Lynn was found to be the most effective anticalculogenic agent. Musa sapiens and dolicos biflorus were found to have no significant effect in inhibiting crystal growth. The kidneys of rats on calculogenisation showed different grades of crystals in the glomerulus and interstitial tissues, extrusion of the crystals into the tubular lumen, collodisation and tissue inflammatory cell infiltration. Scoparia dulcis Lynn exhibited maximum protector effect against the changes of calculogenisation. Musa sapiens and dolicos biflorus had only minimal effect in preventing crystal deposition, inflammatory cell infiltration and other changes of calculogenisation. SEM was found to be effective in assessing the effect of drugs on crystal growth morphology and tissue histology.

The title compound, [Ca(C4H4O6)].4H2O, calcium tartrate tetrahydrate, is a new triclinic centrosymmetric form identified in rat kidney calculus. The crystal structure was determined from powder and single-crystal X-ray diffraction. The four water molecules belong to one square face of the Ca-atom coordination (a square antiprism), the four O atoms of the second square face coming from two tartrate anions, building infinite chains alternating Ca atom polyhedra and tartrate anions along a, with the chains cross-linked by a network of hydrogen bonds.

Background and Purpose: Numerous factors influence the treatment result and efforts for stone removal. To summarize the important factors, our aim was to formulate a general mathematical expression of the stone-removal procedure., Materials and Methods: A mathematical expression (stone treatment index; STI) was developed for patients who became stone free (SF): STI(SF) = (Num(SF) x mean(sqrt [SA] x HI) x meanAge(R) x meanBMI(R) x (1 + meanNum(ANA)) divided by Num(SESSIONS) + Num(AUX) + Num(ANE) + Num(COMP). The variables included were the number of stone-free patients (Num(SF)), mean stone burden expressed as the product of the square root of the stone surface area (SA(1/2)) and the hardness index (HI), the mean age ratio (Age(R) = age/50), the mean body mass index (BMI) ratio (BMI(R) = BMI/25), the mean number of patients with anatomic abnormalities (Num(ANA)), the total number of treatment sessions (Num(SESSIONS)), the number of auxiliary procedures (Num(AUX)), the number of procedures requiring general or regional anesthesia (Num(ANE)), and the number of complications (Num(COMP)). A similar index was calculated for patients who were either stone free or had residual fragments < or =4 mm (STI(DIS)). For conclusions on efficacy, these indices were compared with optimal and total values of STI. The STI was calculated for 450 patients with renal and 374 patients with ureteral stones treated with shockwave lithotripsy as the primary procedure. In patients with a known BMI, this factor was used; otherwise, the BMI(R) was set to 1., Results: When STI was compared with the efficiency quotient (EQ) for stones in different size intervals, STI was less sensitive to variations in stone burden. The quotients STI(SF)/STI(OPTIMAL) corresponded roughly to EQ but might be more informative because of the inclusion of factors for anesthesia and complications., Conclusion: With or without related estimates, the STI might be useful for summarizing stone-removal procedures in groups or individual patients with urinary-tract stones. The STI might be helpful because it encompasses the factors of importance for conclusions about the treatment procedure, such as the hardness of the stone, the age and body size of the patient, and the presence of anatomic abnormalities likely to influence the result. The STI might be used advantageously for comparison of different equipment and treatment strategies.

Identification of chemical constituents of calculus is important in the diagnosis and management of urolithiasis. The compositional variability of uroliths has different etiologies and requires various modes of treatment and prophylaxis. In the present study, we report the chemical compositional analyses of calculi recovered from buck and bullock by X-ray diffraction (XRD) and energy dispersive X-ray (EDX) techniques and ultra-structure examination by scanning electron microscopy (SEM). XRD and EDX investigations conclusively established the chemical compositions of urinary calculi under investigation. The calculus from buck (sample I) had calcium oxalate monohydrate, a dominant salt phase and magnesium compound in significant amount. The calculus from bullock (sample II) had magnesium ammonium phosphate phase, with significant amount of calcium in apatite form and K+ ions. SEM study at higher magnification (X 1000) showed bipyramidal crystals in external zones of urolith (sample I). The struvite apatite calculus showed that basic unit of structure was lamination and the laminitis appeared to be made up of fine granules and high porosity. The bio-mineralization process of calculus formation was also studied, with a view to take preventive and therapeutic measures for amelioration of urinary stone diseases in animals and humans.

Purpose: To measure the hardness of two types of urinary stones: calcium oxalate monohydrate (COM; CaC2O4 . H2O) and dicalcium phosphate dihydrate (brushite; CaHPO4 . 2H2O)., Materials and Methods: The composition of 28 calcium oxalate monohydrate and 22 brushite stones was characterized by infrared spectroscopy (FT-IR). Stone specimens were embedded in crystallographic resin, polished, and subjected to indentation tests using a Vickers tester. The hardness was calculated from measuring the diagonal lengths of the residual indentation on the specimen using the appropriate equation., Results: The COM stones showed hardness values ranging from 15.3 to 64.2 HV with a mean of 35.8 +/- 13.3, while brushite stones ranged from 10.1 to 46.1 HV with a mean of 26.5 +/- 15.1. The results of ANOVA showed that there were significant differences (P < 0.05) between the two stone types., Conclusions: Calcium oxalate monohydrate stones exhibited greater hardness than brushite stones when assessed with Vickers studies.

Monoenergetic x-ray diffraction (XRD) analysis is an established standard for the assessment of urinary stone composition. The inherent low energy of x-rays used (8 keV), however, restricts penetration depth and imposes a requirement for small powdered samples. A technique capable of producing detailed information regarding component structural arrangements in calculi non-destructively would provide clearer insights into causes of formation and subsequent growth and allow the selection of more appropriate courses of therapy. We describe a new method based on the detection of coherent scatter (CS) in stone components using polyenergetic x-rays (70 kVp) from diagnostic equipment. While the higher energy allows the analysis of intact calculi, the polyenergetic source causes an angular broadening of measured CS patterns. We show that it is possible to relate the polyenergetic (CS) and monoenergetic (XRD) measurements through a superposition integral of the monoenergetic XRD cross-section with a function representative of the polyenergetic spectrum used in CS. Experimentally acquired diffractometry cross-sections of the seven major urinary stone components were subjected to this operation, revealing good agreement of diffraction features with CS. Therefore, our CS analysis is sensitive to stone component structure, similar to conventional XRD analysis. This indicates that CS analysis can be used as a basis to classify urinary calculi by composition. The potential of identifying stone components non-destructively was demonstrated from a tomographic CS analysis of a stone-mimicking phantom. Tomographic composition maps were generated from CS patterns, showing the structural arrangement of multiple stone components within the phantom. CS analysis has the ability to detect components in the presence of many others. The ability to perform CS measurements in intact calculi would allow for the identification of stone structures critical to patient metaprophylaxis.

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.

In our laboratory more than 100,000 urinary calculi have been analysed since 1972. Amongst this huge sample, 15 specimens originating from a total of eight patients were observed showing similar characteristics but escaping unambiguous identification with any of the substances that have been described so far in urinary concrements. Therefore, the unknown substance was submitted to a more extended analytical regimen. Structural analysis by x-ray crystallography turned out to be most successful, identifying the unknown material as uric acid monohydrate. Uric acid monohydrate crystallizes in the monocline space group P2(1)/c. Within the crystal, uric acid and water molecules form continuous layers by hydrogen bonds. This is in contrast to uric acid in its water free and its dihydrate forms, which both crystallize by forming 3-dimensional networks To the best of our knowledge , the existence of a monohydrate form of uric acid has not been reported so far. Accordingly, this is the first report on uric acid monohydrate as a urinary stone component. The frequency of only 0.015% in our survey indicates that uric acid monohydrate is rarely the main component in concrements, in contrast to uric acid and uric acid dihydrate with frequencies of 10% and 6%, respectively. The infrared spectrum of uric acid monohydrate is very similar to that of the other crystal forms of uric acid. Because of this similarity and its low frequency, uric acid monohydrate may have been overlooked as a component of urinary concrements. X-ray diffraction allows for better differentiation in routine stone analysis. All samples of uric acid monohydrate were found by solid state NMR spectroscopy to be highly contaminated by amorphous material. This material consisted of long aliphatic chains reminiscent of lipids and fatty acids, respectively. Concrements consisting of other forms of uric acid or urate lacked this amorphous component. Therefore, a role of this aliphatic material has to be taken into consideration when discussing the conditions that may favour the rare formation of concrements from uric acid monohydrate. As for as the metabolic situation of the affected patients is concerned, no common peculiarities became evident by a retrospective survey.

For this study, 25 samples of urinary stones were chosen from different provinces in Iraq as representative sampling localities. These samples of urinary stones were collected to represent kidney, urate, and bladder stones. The main objectives of this study are to try to shed some light on the possibilities of tracking down the effective environmental factors that determine the mineralogical and chemical composition of these stones. The stones were examined using several techniques, the most important of which was the use of the X-ray diffraction (XRD) technique to determine the mineralogical composition of these stones. A scanning electron microscopy (SEM) test was conducted to determine the crystallographic forms and structures for the minerals forming these stones. Optical properties of these minerals were studied using a polarizing microscope. All these techniques revealed that the calcium oxalate, represented in Whewellite mineral, is the most dominant type of these stones, in addition to other minerals such as Hydroxy apatite, Struvite, and Uricite. Dittmarite was pointed out for the first time ever in some samples. This mineral has not been determined in any previous study worldwide. Considering the results of mineralogical and chemical examinations of the urinary stones in question, and the statistical information gathered from the Iraqi Health Ministry, statistical analyses were applied. The ratio of male-female cases in this study happened to be 4:1, which was higher than the ratio in the years 1988-1989 and 1993-1994, 2:1; 3:1 respectively. The highest percentage of the cases was in the 15-50 age group, which is considered as the most productive years of human lifetime. This study showed that one of the most significant factors was that the mineralogical variation of urinary stones in some Iraqi provinces was due to geographical differences, which reflect the variation in lithogenic factors and also climatological factors. Other factors may be socioeconomic, genetic, physiological, and pathological, which remain the important factors in forming urinary stones.

Background: The purpose of the present paper was to study the spectrum of stone composition of upper urinary tract calculi by X-ray diffraction crystallography technique, in patients managed at All India Institute of Medical Sciences., Methods: Between 30 April 1998 and 31 March 2003, a total of 1050 urinary calculi (900 renal, 150 ureteric) were analyzed. The stone fragments were collected after extracorporeal shock-wave lithotripsy, or retrieval by endoscopic (percutaneous nephrolithotomy, ureterorenoscopy), laparoscopic and various open surgical procedures. The structural analysis of the stones was done using X-ray diffraction crystallography., Results: Four types of primary and three secondary X-ray diffraction patterns were obtained. The primary patterns were as follows. Pattern A, well organized crystalline structure; pattern B, moderately organized crystalline structure; pattern C, poorly organized crystalline structure; pattern D, very poorly organized crystalline structure. The three secondary patterns mainly highlighted the mixed variety of stones. These patterns were further analyzed and compared with standard X-ray diffraction (powder) photographs. Of the 1050 stones analyzed, 977 (93.04%) were calcium oxalate stones, out of which 80% were calcium oxalate monohydrate (COM) and 20% were calcium oxalate dihydrate (COD). Fifteen were struvite (1.42%) and 19 were apatite (1.80%). Ten were uric acid stones (0.95%) and the remaining 29 (2.76%) were mixed stones (COM + COD and calcium oxalate + uric acid, calcium oxalate + calcium phosphate, and calcium phosphate + magnesium ammonium phosphate). A total of 89.98% of staghorn stones were made of oxalates (COM/+COD) and only 4.02% were struvite., Conclusion: Urinary stone disease in the Indian population is different from that in Western countries, with a larger percentage of patients having calcium oxalate stones, predominantly COM. Also, the majority of staghorn stones (89.98%) were made of oxalates.

Renal tubular epithelium is the major target for oxalate induced injury, and sustained hyperoxaluria together with CaOx crystal formation/deposition may induce renal tubular cell damage and/or dysfunction. This may express itself in cell apoptosis. To evaluate the possible protective effects of certain agents (vitamin E, potassium citrate, allopurinol, verapamil and MgOH) on the presence and the severity of apoptotic changes caused by hyperoxaluria on renal tubular epithelium, an experimental study in rabbits was performed. Seventy rabbits were divided into seven different groups (each group n = 10): in group I severe hyperoxaluria was induced by continuous ethylene glycol (0.75%) administration started on day 0 and completed on day 14. Histologic alterations including crystal formation together with apoptotic changes (by using the TUNEL method) were evaluated on days 21 and 42, respectively. In the remaining experimental groups (groups II-VI), animals received some agents in addition to the induction of hyperoxaluria in an attempt to limit apoptotic changes. Group VII) animals constituted the controls. Kidneys were examined histopathologically under light microscopy for the presence and degree of crystal deposition in the tubular lumen. The percentage of apoptotic nuclei in the control group was significantly different from the other group animals (2.9-2.4%) in all study phases (P < 0.05). Apart from potassium citrate and allopurinol, the other medications seemed to prevent or limit the formation of apoptotic changes in renal tubular epithelium during the early period (day 21). The percentage of positively stained nuclei in animals undergoing potassium citrate medication ranged from 24.3% to 28.6%, with an average of 27.1%. This was 18.4% in animals receiving allopurinol. On the other hand, animals receiving magnesium hydroxide (MgOH), verapamil and vitamin E demonstrated limited apoptotic changes (11.2, 9.7, 8.7%, respectively) during this phase(P < 0.05). In the long-term (day 42), the animals receiving allopurinol and vitamin E showed a decrease in the percentage of the positively stained nuclei (13.5% and 8.3%, respectively). Animals in the other groups showed an increase in the number and percentage of apoptotic cells. Although, there was a significant decrease in the mean values of apoptosis in animals receiving vitamin E (8.7%-8.3%) and allopurinol (18.4%-13.5%) (P < 0.05), animals on verapamil, MgOH and potassium citrate medication had an increase in these values or the change was not found to be significant. In the light of our findings and results from the literature, it is clear that that both hyperoxaluria and CaOx crystals may be injurious to renal epithelial cells. Apoptotic changes observed in renal tubular epithelial cells induced by massive hyperoxaluria might result in cell degradation and may play a role in the pathologic course of urolithiasis. Again, as demonstrated in our study, the limitation of both crystal deposition and apoptotic changes might be instituted by some antioxidant agents as well as urinary inhibitors. Clinical application of such agents in the prophylaxis of stone disease might limit the formation of urinary calculi, especially in recurrent stone formers., (Copyright 2004 Springer-Verlag)

The chemical constituents of 34 feline urinary calculi and five urethral plugs were analysed by infrared spectroscopy. The analysis revealed that 18 (52.9 per cent) of the calculi contained magnesium ammonium phosphate hexahydrate (struvite) as the major component; 10 (29.4 per cent) contained complex ammonium urates (three of them also containing calcium phosphate, mainly on the surface); three were composed of calcium phosphates and three were composed mainly of calcium oxalate mono and dihydrates. The urethral plugs were composed primarily of struvite, but also contained large amounts of organic matter. The examination of 16 selected samples by scanning electron microscopy and electron dispersive x-ray analysis revealed that their crystalline structures were similar to those of canine stones.

A single team has reported isolation of nanobacteria in human and bovine blood products, as well as, more recently, kidney stones. This has raised controversy. To confirm the data, we searched for nanobacteria from 10 aseptically removed upper urinary tract (UUT) stones. We used scanning electronic microscopy (SEM) with four stones and culture of stones on either 3T6 fibroblast monolayers or liquid RPMI medium. Detection of nanobacteria was made with a commercially available monoclonal antibody, 16S ribosomal DNA amplification with specific primers, and transmission electronic microscopy (TEM) of inoculated cells. SEM showed nanoparticles in four of four UUT stones similar to those recently described. TEM of inoculated 3T6 cell monolayers has shown transient intracytoplasmic vacuolar formations containing 200- to 500-nm particles in 3 of 10 cell cultures. Gimenez staining, Hoechst staining, and specific monoclonal immunofluorescence failed to reveal nanobacteria. Finally, we could not grow Nanobacterium sp. microorganisms by the techniques described. Although with SEM, we observed nanoparticles morphologically similar to nanobacteria, we failed to isolate Nanobacterium sp. microorganisms in culture and to prove the bacterial nature of these nanoparticles in stones.

Sulfosulfuron, developed as a herbicide, caused increased microcrystalluria and the formation of urinary tract calculi when fed to male and female rats in a chronic 2-year study at doses of 5,000 ppm and 20,000 ppm. Hyperplasia was also seen in urinary bladders at 5,000 ppm and 20,000 ppm, almost exclusively in the presence of observable calculi/microcalculi. Urinary bladder tumors were found in 2 females in the 5000 ppm group, both in the presence of calculi. No increased microcrystalluria, calculi, or tumors were found at doses of 500 ppm and lower. In the current study, 5 groups of male Sprague-Dawley rats were fed sulfosulfuron at doses of 50, 500, 5,000, and 20,000 ppm for 10 weeks. Ten animals were co-administered 5,000 ppm sulfosulfuron with 12,300 ppm NH4Cl to determine if inhibition of the formation of calculi would prevent any urothelial effects of treatment with sulfosulfuron. Ten animals in the control group and in the high-dose sulfosulfuron group were fed only basal diet for an additional 10 weeks to determine if the effects of sulfosulfuron on the bladder epithelium were reversible. There was an increased incidence of microcrystalluria observed at 5,000 and 20,000 ppm. There was no increase in microcrystalluria observed in the urine of rats co-administered sulfosulfuron and NH4Cl. Urinary bladder calculi were found in the bladder of 1 animal fed 20,000 ppm. Examination by light microscopy showed diffuse papillary/nodular hyperplasia of the bladder epithelium in this animal. No increased microcrystalluria was observed after withdrawal of the chemical from the diet and the bladder epithelium was normal by light microscopy. The hyperplastic effects associated with the feeding of high doses of sulfosulfuron occur only with the appearance of urinary tract calculi. Based on these results and anatomical differences between rats and humans, it may be concluded that the hyperplastic and carcinogenic effects of sulfosulfuron in rats are high-dose, threshold phenomena that are not likely to occur in humans under environmentally relevant exposures.

Objective: To elucidate the ultrastructural details of calcium oxalate-containing urinary calculi from dogs. Sample Population-38 specimens selected from a collection of 8,297 oxalate-containing urinary calculi from dogs: 22 specimens composed of calcium oxalate (calcium oxalate monohydrate [COM], calcium oxalate dihydrate [COD], or COM and COD) and 16 specimens composed of calcium oxalate with amorphous calcium phosphate., Procedure: Analyses of specimens included use of plain, reflected, and polarized light microscopy, X-ray diffractometry, scanning electron microscopy (SEM) with backscattered electron (BSE) imagery, and electron microprobe analysis., Results: Four texture types were observed in calcium oxalate calculi; 4 texture types of calcium oxalate-calcium phosphate-mixed calculi were recognized. Texture types were delineated through differences in calcium oxalate crystal sizes, which were affected by urine supersaturation and abundance of crystal nucleation sites. Segregation of calcium oxalate from calcium phosphate indicated they do not precipitate under the same conditions. Deposition of calcium phosphate between calcium oxalate crystals decreased the volume of pore spaces within calculi. Porosity was observed along boundaries between COM and COD. Minute pores increased the surface area of calculi exposed to urine, and this increase in liquid-solid interface promotes interaction of crystals with the surrounding urine., Conclusions and Clinical Relevance: Calcium oxalate urolithiasis is of major concern, because it is often a recurrent disease among dogs, principally treated by surgical removal of calculi, with few effective dissolution strategies. Understanding the ultrastructure and mineralogic content of calcium oxalate and its association with amorphous calcium phosphate is a step toward the solution of this increasingly important medical problem.

The careful analysis of cystine calculi may be important to detect the presence of other urinary alterations (such as hyperuricosuria, hypercalciuria, hyperoxaluria or bacterial infections) that coexist with cystinuria in many patients. For this reason, in the present study, 14 human and 17 canine cystine uroliths have been studied by infrared spectroscopy (IR), scanning electron microscopy (SEM), and energy dispersive X-ray analysis (EDX). According to the infrared analysis, most of the human and canine stones were composed of nearly pure cystine. However, in these calculi of apparently pure cystine, the study by SEM and EDX showed in many cases the presence of small amounts of calcium apatite. The infrared study of several samples heated at 750 degrees C confirmed the presence of phosphate, when it was difficult to detect this component in the spectra of the original samples owing to band overlapping. Other components detected in varying proportions in cystine calculi were magnesium ammonium phosphate hexahydrate (struvite), calcium hydrogen phosphate dihydrate (brushite), calcium oxalate (mono and/or dihydrate) and, in one case, a drug (oxolinic acid).

Purpose: We test the hypothesis that holmium:YAG lithotripsy efficiency varies with optical fiber size and energy settings (energy density)., Materials and Methods: The 272, 365, 550 and 940 microm. optical fibers delivered 1 kJ. total holmium:YAG energy to calcium oxalate monohydrate calculi at energy output/pulse of 0.2 to 1.5 J. Stone mass loss was measured for each fiber energy setting. Stone crater width was characterized for single energy pulses. Fiber energy outputs were compared before and after lithotripsy., Results: Stone mass loss correlated inversely with optical fiber diameter (p <0.05). Stone loss correlated with energy/pulse for the 365, 550 and 940 microm. fibers (p <0.001). The 272 and 365 microm. fibers yielded equivalent stone loss at 0.2 and 0.5 J. per pulse. At energies of 1.0 J. per pulse or greater the 272 microm. optical fiber was prone to damage, and yielded reduced energy output and stone loss compared to the 365 microm. fiber (p <0.01). Stone crater width for single pulse energies correlated with energy settings for all fibers (p <0.001)., Conclusions: Lithotripsy efficiency with the holmium:YAG laser depends on pulse energy output and diameter of the optical delivery fiber, implying that lithotripsy efficiency correlates with energy density. The 365 microm. fiber is indicated for most lithotripsy applications. The 272 microm. fiber is susceptible to damage and inefficient energy transmission at energies of 1.0 J. per pulse or greater. The 272 microm. fiber is indicated at energies of less than 1.0 J. per pulse for small caliber ureteroscopes or when maximal flexible ureteroscope deflection is required.

The number of nucleolar organizer regions (NOR) of human bladder cancers was evaluated at the light- and electron-microscopic level. The average number of argyrophilic NOR (AgNOR), stained by the one-step silver colloid method, was measured in benign and malignant urothelial cells in the human urinary bladder using a light microscope. The average number of nucleolar fibrillar centers (FC) per nucleus was also calculated by quantitative ultrastructural morphometry in the specimens from the same patients. Statistical evaluations revealed that the average number of AgNOR per nucleus was significantly correlated with the elevation of tumor grade and stage (p < 0.05). An average FC number per nucleus also increased in association with tumor grade and stage (p < 0.05). Although the average number of FC was 5.6 times higher than that of AgNOR, the correlation between the average number of FC and AgNOR was statistically significant. In conclusion, these results suggested that the silver staining method was a useful and convenient tool for the evaluation of the differentiation and invasive potential of bladder cancer cells at the light-microscopic level.

A uric acid concretion formed round a catheter (JJ stent) in the bladder and removed intact from the body together with the catheter was studied using an electron scanning microscope. The concretion was composed of anhydrous uric acid, some uric acid dihydrate (< 5 wt.%) and individual particles of calcium oxalate monohydrate. The stone interior was porous with frequent occurrence of differently sized cavities that were either empty or partially filled with particles of uric acid and/or calcium oxalate monohydrate. Calcium oxalate particles were not of crystalluria origin but developed in the cavity. The succession of processes leading to the stone formation was deduced from its inner structure. The stone was formed due to a crystalline growth with minor, if any, participation of sedimentation. The estimated average rate of the calculus development, 2 x 10(-9) m/s, confirms the predominant role of crystalline growth in stone formation and indicates a relatively low urinary supersaturation with respect to uric acid prevailing during the period of calculogenesis.

Stones with different compositions respond differently to shock wave lithofragmentation. Likewise, the various lithotrity systems used may have different effects on the stones. To determine the relationships between stone composition and their fragmentation patterns, we conducted an in vitro study using endoscopy, magnifying glass, light microscope and scanning electron microscope on fragments obtained after lithotrity of 60 pure stone with different compositions: calcium oxalate monohydrate and dihydrate (OXMH and OXDH), phosphocarbonate (PC), ammonium magnesium phosphate (AMF) and uric acid (UA). Fragmentation was carried out with 4 different lithofragmenting sources (electrohydraulic, piezoelectric, ultrasound and pulse laser). No morphologic differences in the fractures induced by the various lithofragmenting sources were demonstrated. OXMH and UA stones basically break up by intercrystalline fracture and splitting of their concentric plates. OXDH breaks up mainly by intercrystalline fractures aided by the fibrillar organic material and phosphocarbonates found in the intercrystalline spaces. Fragmentation in infective stones (AMF and PC) occurs across the intercrystalline surfaces and by intracrystalline fracture. Ammonium urate fragments break up by intracrystalline fractures that run across the equatorial plane of its characteristic acicular microspheres.

The chemical composition and microstructure of seven uroliths and four urinary sediment samples associated with the feeding of high-level cottonseed meal diet to buffalo calves were examined by chemical qualitative analyses, scanning electron microscopy (SEM), X-ray diffraction, and X-ray energy dispersive spectrometry (EDS). Struvite was a major component of kidney stones and of some bladder stones. The kidney stone sample appeared cracked under low power under SEM, aggregated into tiny balls under high power, and as a blade-like structure under even higher power. The bladder stone samples appeared finely granular or granular with various forms of prismatic crystals. The urinary sediments were prismatic crystals, with granules. The newly found prismatic crystals, which were rich in potassium and similar to struvite in crystal structure, were identified as potassium magnesium phosphate (KMgPO4.6H2O) in some bladder stones and urinary sediments. However, crystals which contained Mg and P only, which had been used for struvite identification, were not found by EDS examination in urinary sediments from fresh urine samples of buffalo calves fed the high-level cottonseed meal diet.

The ultrastructure of the organic matrix of demineralized urinary stones was examined by standard transmission and scanning electron microscopy as well as after malachite green-glutaraldehyde fixation. Crystal ghosts of both calcium oxalate and calcium phosphate were made of amorphous material and were dispersed in a matrix containing amorphous, fibrillar, and membranous substances. Malachite green positive material was seen to be associated with the ghosts, as well as with the membranous and fibrillar components of the organic matrix. Calcium oxalate and calcium-phosphate crystals, induced in human urine in vitro were also found to be associated with an organic matrix containing lipids and proteins. It is suggested that the intimate association between crystals and lipids is a result of the involvement of cellular membranes in the nucleation of these crystals.

Objective: To elucidate the ultrastructural details of struvite-containing urinary calculi from dogs., Sample Population: 38 specimens were selected from a collection of approximately 13,000 canine urinary calculi: 18 of these were composed entirely of struvite, and 20 consisted of struvite and calcium phosphate (apatite)., Procedure: Qualitative and quantitative analyses of specimens included use of plain and polarized light microscopy, x-ray diffractometry, scanning electron microscopy with backscattered electron imagery, x-ray fluorescence scans, and electron microprobe analysis., Results: 4 textural types were recognized among struvite calculi, and 4 textural types of struvite-apatite calculi were described. Evidences of calculus dissolution were described from 4 calculi studied., Conclusions: The presence of small, well interconnected primary pores in struvite-containing urinary calculi from dogs appears to be a significant factor in determining the possible interaction of calculi with changes in the urine composition. The progress of dissolution from the calculus surface to the calculus interior appears to be largely affected by the primary porosity originally present between crystals forming the calculus framework. Apatite was observed to be more resistant to dissolution than struvite., Clinical Relevance: The prevalence of fine concentric laminations having low porosity, and the common occurrence of apatite among struvite-containing urinary calculi from dogs may be 2 reasons why the efficacy of dietary and medicinal manipulations in dissolving urinary calculi is greater among cats than it is among dogs.

Crystalluria results from oversaturation of urine with crystallogenic substances. However, oversaturation may occur as a result of in vivo and in vitro events. Therefore, care must be used not to overinterpret the significance of crystalluria. Evaluation of urine crystals may aid in (1) detection of disorders predisposing cats to urolith or matrix-crystalline urethral plug formation; (2) estimation of the mineral composition of uroliths or urethral plugs; and (3) evaluation of the effectiveness of medical protocols initiated to dissolve or prevent urolithiasis.

We investigated the effects of weak to moderate urease hydrolysis by optional urease-positive microorganisms in an artificial urine model enriched with calcium phosphate and calcium oxalate in respect of calcium stone formation. The incubation experiments were performed using a discontinuously running fermenter device to simulate the urinary system. The kinetics of cell division rates, pH and ammonium ion production were measured and correlated to crystallite appearance in the incubation medium. Qualitative analyses of the sediments revealed apatite. Investigations using light microscopy and scanning electron microscopy (SEM) confirmed the matrix effect of bacterial glycoproteins. It was shown that initiation of calcium oxalate stone formation is in all probability equally determined by matrix effects and by heteronuclear crystallization if the urinary tract is infected by optional urease-positive bacteria. When urinary inorganic phosphate is present, calcium phosphate nidi are always initially formed, and may subsequently be coated by calcium oxalate.

It is generally recognized that calcium oxalate crystal formation in urine is induced by heterogeneous nucleation. However, there is no consensus as to the nature of the nucleation substrate. Evidence is provided in this paper that membranous cellular degradation products are the most likely candidates because they: (1) are ubiquitous in urine and urinary stones; (2) are found in close association with crystal deposits in the kidneys; and (3) can induce nucleation of crystals from a metastable solution of calcium oxalate in vitro and metastable urine in vivo.

Urinary calculi from 17 horses with urolithiasis were examined to study their mineral content and ultrastructure. Among the analytic methods used were X-ray diffractometry, scanning electron microscopy, and electron microprobe analysis. The calculi initially were observed by use of a stereoscopic dissecting microscope and generally were found to have nodular surfaces surrounding a banded or granular-to-chalky interior. Observation by scanning electron microscopy revealed an intricate pattern of irregularly concentric, fine bands and spherules. These had a round, finely banded, globular texture formed by precipitation of ultrafine-grained radiating crystals. The original pore spaces (ie, between spherules, between bands and spherules, or between crystal generations) could be observed as primary porosity. Precipitation and dissolution of these urinary calculi were observed to be spontaneous processes, which can occur simultaneously within an individual calculus. Another prominent feature of the ultrastructure was secondary porosity (spontaneous dissolution) which, in its incipient stages, appeared to be site-selective (ie, some bands appeared to be more susceptible to development of pinpoint porosity). Textures indicative of dissolution were observed not only on the calculus surface, but within the calculus interior as well. Areas that had more advanced stages of dissolution, resulting in increased secondary porosity, also were observed. All 17 samples of the study were found to be composed of calcium carbonate in the form of the mineral calcite, although minor quantities of 2 other polymorphs of calcium carbonate, minerals vaterite and aragonite, also were encountered. Vaterite was observed in 5 of the samples, whereas aragonite was found in 1 sample. Strontium and sulfur were observed as trace elements in 3 of the calculi, whereas magnesium was present in all calculi. Magnesium was observed to substitute for calcium within the calcite crystal lattice in larger quantities than those of strontium or sulfur. Magnesium K alpha X-ray dot maps generated by use of an electron microprobe analyzer indicated that the distribution pattern of magnesium appeared to closely follow layer-by-layer growth of the calculus. Magnesium distribution also appeared to be related to porosity development. In samples where preferential dissolution was observed, the more porous areas had higher magnesium content. Quantitative chemical analyses, using the electron microprobe analyzer, confirmed these observations. Association of the magnesium distribution pattern to the primary growth texture of the calculus indicated that magnesium content of the calculus varied during the formation process. This also indicated that changes in urine chemical analytes may be reflected in composition of the calculi formed.(ABSTRACT TRUNCATED AT 400 WORDS)

Struvite (MgNH4PO4.6H2O) calculi are a common complication of Proteus mirabilis urinary tract infections. Although urease is a major virulence factor in calculus formation, the polysaccharide capsule (CPS) of this organism also enhances struvite crystallization and growth in vitro (L. Clapham, R. J. C. McLean, J. C. Nickel, J. Downey, and J. W. Costerton, J. Crystal Growth 104:475-484, 1990). We obtained purified CPS, of known structure and varying anionic character, from P. mirabilis ATCC 49565 and several other organisms. Artificial urine was added to CPS, and the pH was elevated from 5.8 to 8.5 by the addition of urease or titration with 0.25 M NH4OH to induce struvite crystallization. Crystallization was measured by particle counting (Coulter counter), and the morphology (crystal habit) was examined by phase-contrast microscopy. In the presence of partially anionic P. mirabilis CPS, struvite formation occurred at a lower pH than in the absence of CPS or in the presence of other neutral, partially anionic, or anionic CPS. At pH 7.5 to 8.0, significantly more struvite crystals formed in the presence of P. mirabilis CPS than under other experimental conditions. With the exception of one polymer (curdlan) which did not bind Mg2+, enhancement of struvite formation by CPS polymers was inversely proportional to their Mg2+ binding ability. We speculate that the structure and partial anionic nature of P. mirabilis CPS enable it to enhance struvite formation by weakly concentrating Mg2+ ions during struvite crystal formation. This illustrates a new virulence aspect of bacterial CPS during infection.

Urinary calculi are often small (< 5 mg) and can be composed of many very different constituents. The highly variable composition has lead to the development of many different methods of calculi analysis. In general the analytical methods can be divided in chemical and physical methods. Chemical methods are destructive and need several mg of sample. The smallest stones can not be analyzed with chemical methods. Qualitative and semi-quantitative chemical analysis methods are not accurate and can lead to clinical significant errors. Quantitative, instrumental, chemical analysis is accurate and makes no clinical significant errors, but these consume too much sample. None of the chemical methods of stone analysis makes it possible to distinguish the mineral constituents with similar chemical composition, e.g. the calcium phosphates, from one another and they usually can not identify "unexpected" stones. Physical methods need less sample than chemical methods and physical methods can distinguish the different minerals in the stones. Thermal analysis can give quantitative results but needs relatively much sample. Optical methods can analyze a few grains of stone but gives only qualitative results. Both methods can not be used to analyze an "unexpected" stone. Best suited methods for the analysis of calculi are X-ray diffractometry and IR spectroscopy. X-ray diffractometry can detect crystalline minerals in low concentrations. IR spectroscopy can detect both crystalline and amorphous minerals. Both methods can be used for semi-quantitative stone analysis and can analyze less than one mg of stone sample. X-ray diffractometry and IR spectroscopy are also the best methods to identify "unexpected" stone constituents.