Calcium oxalate urolithiasis in juvenile dogs.

BACKGROUND: The features of juvenile-onset calcium oxalate urolithiasis in dogs have not been previously reported. METHODS: Calcium oxalate urolith submissions to the Minnesota Urolith Center between 2012 and 2016 were analyzed to identify those originating from juvenile (≤2 years, n = 510) or mature (7-9 years, n = 39,093) dogs. Breed, sex, urolith salt type and urolith location were compared between groups. Breeds represented in both groups were also compared with respect to sex, urolith salt type and urolith location. RESULTS: French (odds ratios [OR] = 14.7, p < 0.001) and English (OR = 14.3, p < 0.001) Bulldogs were overrepresented in juvenile submissions. All juvenile French and English Bulldogs were male. Across all breeds, juvenile dogs were more likely to be male (89%, p < 0.001) than mature dogs (79%). Juvenile dogs were also more likely to form dihydrate stones compared to mature dogs (33% versus 14%, respectively; p < 0.001). Breed differences were discovered in sex, urolith salt type and stone location. CONCLUSIONS: French and English Bulldogs comprise a greater proportion of juvenile calcium oxalate urolith submissions than expected based on their rarity in mature submissions. Inherited risk factors, particularly X chromosome variants, should be investigated due to the strong breed and sex predispositions identified.

Clinical-grade Garcinia cambogia extract dissolves calcium oxalate crystals in Drosophila kidney stone models.

OBJECTIVE: Kidney stone formers have a high rate of stone recurrence after kidney stone removal surgery and there is no effective medication for treatment. Hydroxycitric acid (HCA), which is the major component of Garcinia cambogia extract, can dissolve calcium oxalate crystals in vitro, suggesting that Garcinia cambogia could be used to treat calcium oxalate kidney stone. In this study, we used the Drosophila kidney disease model to evaluate the effect of Garcinia cambogia on the prevention and removal of calcium oxalate stones in vivo. MATERIALS AND METHODS: Flies were reared in fly food containing different concentrations of GCE for one week. The effect of GCE on preventing the formation of calcium oxalate stone was examined. WT and v-ATPase gene RNAi knockdown flies were reared in fly food with 0.3% NaOx for one week, then fed different concentrations of GCE for one week. The effect of GCE on the removal of calcium oxalate stone was examined. RESULTS: Garcinia cambogia extract dissolves calcium oxalate crystals from Malpighian tubules in both genetic and non-genetic Drosophila kidney stone models compared to citric acid. Hydroxycitric acid also directly dissolves calcium oxalate crystals in Drosophila Malpighian tubules ex vivo. CONCLUSIONS: Garcinia cambogia extract removes calcium oxalate kidney stones from Drosophila Malpighian tubules via directly dissolving calcium oxalate stones by HCA. Our study strongly suggests that clinical-grade Garcinia cambogia extract could be used to treat patients with nephrolithiasis in the future.

[Study on irritation of calcium oxalate crystal in Araceae plants].

OBJECTIVE: To validate the irritation effects of calcium oxalate crystal in several herbal drugs which come from Araceae plants. METHOD: Compared the irritation of pure calcium oxalate crystals isolated from the raw rhizome of Typhonium flagelliforme, T. giganteum and Arisaema erubescens and studied the quantity and irritating effect relationship of different concentration suspensions of needle-like calcium oxalate crystals by using the model of rabbits' eyes. RESULT: Calcium oxalate crystals isolated from above three rau rhizome typhonium rhizome showed strong irritation effects on rabbits' eyes. Under the condition of same content of calcium oxalate crystals, there were no difference in irritation effect between the suspensions of raw medicinal materials and pure calcium oxalate crystals. The degree of irritation on rabbits' eyes showed undoubted quantity and irritating effect relationship with the concentrations of Calcium oxalate crystel. CONCLUSION: Calcium oxalate crystal is the irritant component in some herbal drugs which come from Araceae plants.

Isolation from human calcium oxalate renal stones of nephrocalcin, a glycoprotein inhibitor of calcium oxalate crystal growth. Evidence that nephrocalcin from patients with calcium oxalate nephrolithiasis is deficient in gamma-carboxyglutamic acid.

We have determined that the organic matrix of calcium oxalate kidney stones contains a glycoprotein inhibitor of calcium oxalate crystal growth (nephrocalcin) that resembles nephrocalcin present in the urine of patients with calcium oxalate stones and differs from nephrocalcin from the urine of normal people. Pulverized calcium oxalate renal stones were extracted with 0.05 M EDTA, pH 8.0; nephrocalcin eluted in five peaks using DEAE-cellulose column chromatography, and each peak was further resolved by Sephacryl S-200 column chromatography. Four of the five DEAE peaks corresponded to those usually found in nephrocalcin from urine; the fifth eluted at a lower ionic strength than any found in urine. Amino acid compositions and surface properties of nephrocalcins isolated from kidney stones closely resembled those of nephrocalcins isolated from urine of stone-forming patients: they differed from normal in lacking gamma-carboxyglutamic acid residues, and in forming air-water interfacial films that were less stable than those formed by nephrocalcin from normal urine.

Electrophoretic separation and characterization of urinary glycosaminoglycans and their roles in urolithiasis.

Urinary polyanions recovered from the urine samples of kidney stone-formers and normal controls were subjected to preparative agarose gel electrophoresis, which yielded fractions 1-5 in a decreasing order of mobility. In both groups, chondroitin sulfates were identified in the fast-moving fractions and heparan sulfates in the slow-moving fractions. Furthermore, two types of heparan sulfates were identified based on their electrophoretic mobility: slow-moving and fast-moving. The fractionated urinary polyanions were then tested in an in vitro calcium oxalate crystallization assay and compared at the same uronic acid concentration, whereby, the chondroitin sulfates of stone-formers and heparan sulfates of normals enhanced crystal nucleation. Fraction 5 of the normals, containing glycoproteins (14-97 kDa) and associated glycosaminoglycans, were found to effectively inhibit crystallization. Papainization of this fraction in stone-formers revealed crystal-suppressive effects of glycoproteins, which was not seen in similar fractions of normals. It was concluded that glycoproteins could modulate the crystal-enhancing glycosaminoglycans such as chondroitin sulfates of stone-formers but not in normals. The differing crystallization activities of electrophoretic fraction 1 of normals and stone-formers revealed the presence of another class of glycosaminoglycan-hyaluronan. Hence, in the natural milieu, different macromolecules combine to have an overall outcome in the crystallization of calcium oxalate.

[Study on processing mechanism of Pinellia ternate].

OBJECTIVE: Elucidating the detoxification mechanism of the raw Pinellia ternata processed by alum solution or alkaline solution (pH > 12). METHOD: Raw Pinellia ternata was immersed in alum solution and alkaline solution according to Chinese pharmacopoeia. Observed the shape's changing of needle-like calcium oxalate crystals by scanning electro-microscopy. Determinating the contents of calcium oxalate crystals by applying oxidation reduction titration. Measured the irritations of raw P. ternata and various processing products on the model of rabbits'eyes. RESULT: After processed by 8% alum solution prescribed in Chinese pharmacopoeia or 10% sodium carbonate solution, the needle-like shape of raphides in raw Pinellia ternata were changing and the sting barb of raphides were rusted and dissolved, the contents of calcium oxalate crystal in raw Pinellia ternata were sharply declined from more than 1% to less than 0.5%. the decline of contents is relative to the irritation decline of P. ternata on rabbit's eyes. Less than 0.5% calcium oxalate crystals of P. ternata almost had no irritation. CONCLUSION: After processed by 8% alum solution or sodium carbonate solution (pH > 12) , the irritation components in raw P. ternata could be rusted and dissolved, the needle point of raphides was broken, which led to the raphides'content declining and the irritation disappearing. The micro-structures, shapes and contents of calcium oxalate crystals in different medicine plants were not same. These properties of calcium oxalate crystal in India Madder Root and yam et al were very different from those in raw P. ternata.

[Study on irritation of calcium oxalate crystal in raw Pinellia ternata].

OBJECTIVE: Confirm the irritation of needle-like calcium oxalate crystals in raw Pinellia ternata. METHOD: Comparing the irritations of raw P. ternate containing calcium oxalate crystals, the raw P. ternate no containing calcium oxalate crystals, the pure needle-like calcium oxalate crystals isolated from raw P. ternata, the extracts of water and various solvents from raw P. ternate. by using the model of rabbits' eyes. Studying the quantity effect relationship of different concentration suspensions of needle-like calcium oxalate crystal isolated from raw P. ternate on rabbits' eyes. Observing the shape and appearance of calcium oxalate crystals in raw P. ternate and raw India Madder Root by the electro microscope and comparing their irritation degrees with the same contents of calcium oxalate crystals. RESULT: Calcium oxalate crystals in raw P. ternata showed very strong irritation property. Under the same content of calcium oxalate crystals, the irritation of raw P. ternata and pure needle-like calcium oxalate crystals isolated from raw P. ternate had no significant difference. The concentrations of needle-like calcium oxalate crystals were do relative to the degree of irritation on rabbits' eyes and they showed undoubted quantity-effect relationship. CONCLUSION: Calcium oxalate crystal is the irritation component in raw P. ternata.

Calcium oxalate precipitation by diffusion using laminar microfluidics: toward a biomimetic model of pathological microcalcifications.

The effect of mixing calcium and oxalate precursors by diffusion at miscible liquid interfaces on calcium oxalate crystalline phases, and in physiological conditions (concentrations and flow rates), is studied using a microfluidic channel. This channel has similar dimensions as the collection duct in human kidneys and serves as a biomimetic model in order to understand the formation of pathological microcalcifications.

Evidence of formation of glushinskite as a biomineral in a Cactaceae species.

The X-ray diffractometric and infrared spectroscopic investigation of crystalline material isolated from the Cactaceae species Opuntia ellisiana shows the presence of a very complex mineral composition, including whewellite (monohydrated calcium oxalate), opal (SiO2), calcite (CaCO3) and glushinskite (dihydrated magnesium oxalate). This is the first report of the presence of magnesium oxalate in plants.

Epidemiology of canine urolithiasis in the Czech Republic from 1997 to 2002.

OBJECTIVES: To compare data on the epidemiology of canine urolithiasis in the Czech Republic with that from other countries. METHODS: The records from the Centre for Mineralogical Analysis from 1997 to 2002 were reviewed. The data were obtained from mineralogical analysis of 1366 canine uroliths obtained from patients in the Czech Republic. These included 396 females and 629 males. RESULTS: Sixty-eight breeds plus crossbreeds were identified. Eight breeds plus the crossbreeds accounted for 71.3 per cent of all cases. Males were affected more frequently than females (61.4 per cent versus 38.6 per cent). Struvites significantly predominated in females, while in males calcium oxalates, brushites and cystines were the most common stones. Most of the uroliths (48.9 per cent) were 5 mm or less in dimension. By 2001, struvite was the most frequent (38.5 to 44.1 per cent) urolith, followed by calcium oxalate (26.5 to 32.0 per cent). In 2002, calcium oxalate became the most frequent calculus, followed by struvite, mixed calculi and others. CLINICAL SIGNIFICANCE: Comparison of these results with studies by other authors showed that for most of the monitored parameters there was agreement with respect to the proportions of different breeds within the populations of dogs in different geographical areas.

Extraction and estimation of the quantity of calcium oxalate crystals in the foliage of conifer and hardwood trees.

The main goal of this study was to develop a method for the extraction and indirect estimation of the quantity of calcium oxalate (CaOx) in the foliage of trees. Foliar tissue was collected from a single tree of each species (five conifers and five hardwoods) for comparison of extractions in different solvents using 10 replicates per species from the same pool of tissue. For each species, calcium (Ca) and oxalate were extracted sequentially in double deionized water and 2N acetic acid, and finally, five replicate samples were extracted in 5% (0.83N) perchloric acid (PCA) and the other five in 2N hydrochloric acid (HCl); three cycles of freezing and thawing were used for each solvent. Total ions were extracted by microwave digestion. Calcium was quantified with an inductively coupled plasma emission spectrophotometer method and oxalate was eluted and quantified using a high performance liquid chromatography method. This experiment was repeated again with two conifer and two hardwood species using four trees per species, and two analytical replicates for each tree. We report here that, regardless of age of individual trees within a species, time of collection or species type, the third extraction in PCA or HCl resulted in near equimolar quantities of Ca and oxalate (r(2) ≥ 0.99). This method provides an easy estimate of the quantity of CaOx crystals using a small sample of foliar tissue. An additional benefit of PCA is that it precipitates the nucleic acids and proteins, allowing the quantification of several free/soluble metabolites such as amino acids, polyamines, organic acids and inorganic elements all from a single sample extract.

Crystallochemical characterization of calcium oxalate crystals isolated from seed coats of Phaseolus vulgaris and leaves of Vitis vinifera.

Calcium oxalate crystals are a major biomineralization product in higher plants. Their biological function and use are not well understood. In this work, we focus on the isolation and crystallochemical characterization of calcium oxalate crystals from seed coats of Phaseolus vulgaris (prisms) and leaves of Vitis vinifera (raphides and druses) using ultrastructural methods. A proposal based on crystal growth theory was used for explaining the existence of different morphologies shown by these crystals grown inside specialized cells in plants.

First evidence for the presence of weddellite crystallites in Opuntia ficus indica parenchyma.

Calcium oxalate crystallites occur very often in the plants tissues and their role is still poorly known. We report here the experimental protocol leading to the isolation of two forms of calcium oxalate crystallites differing in their hydration level in the parenchymal tissues of Opuntia ficus indica (Miller). Whereas the whewellite crystallites are habitual in all Opuntia species, the weddellite form has never been isolated from these species before, which is probably due to their small size (about 1 microm). We have identified these forms using X-ray diffraction and scanning electron microscopy.

[Calcium oxalate crystals in several kinds of Cinnamon bark].

OBJECTIVE: To probe into the morphology and distribution characteristics of calcium oxalate crystals in several kinds of Cinnamon bark. METHOD: Forty samples of ten different species of Cinamomi Cortex were examined, and the inter-cellular calcium oxalate crystals in the phloem rays were observed by optical microscope and scanning electron microscope. RESULT: It was found that for samples of the same botanical origin, there is little variation in the crystal morphology following certain regularity. The amount, size and ultra-micromorphologic characteristics are influenced by various factors. CONCLUSION: The pattern of morphology and distribution of calcium oxalate crystals may well be an index for identification of the crude drug of Cinnamon bark.

[Study of stinging crystals in tian nanxing].

OBJECTIVE: To test whether the raphides in Tian Nanxing (Pinellia pedatisecta Schott) caused irritation. METHOD: Scan Electron Microscope and Microscope Oberservation; Animal experimental study. RESULT: Through the comparision of unprocessed Tian Nanxing to processed ones which included 36 h, 72 h, 120 h processed samples, the great modifications in the structure of raphides, especially for the ones with barbs in the processed samples was observed with time course study. A further animal experimental study went to show that the rate of change for raphides with barbs existed a dose-reponse relationship to irritation. CONCLUSION: Investigations of the causes of these reactions showed that raphides of calcium oxalate are, at least in part, responsible for the Tian Nanxing's irritation.

[Research on microstructure of caulis herb].

OBJECTIVE: To provide reference for the microscopic identification of caulis herb. METHOD: Systematic arrangement and comparative studies were carried out on the microstructure of medicinal herb of different groups and shapes. RESULT: The rule and characteristics of the microstructure of caulis herb were discussed, and the sorting search list of the microstructure of common caulis herb was established. CONCLUSION: The microstructure characteristics of caulis herb, as the reference of the microscopic identification, are of research value.

Identification of powdered Chinese herbal medicines by fluorescence microscopy, Part 1: Fluorescent characteristics of mechanical tissues, conducting tissues, and ergastic substances.

The light microscope has been successfully used in identification of Chinese herbal medicines (CHMs) for more than a century. However, positive identification is not always possible. Given the popularity of fluorescence microscopy in bioanalysis, researchers dedicated to finding new ways to identify CHMs more effectively are now turning to fluorescence microscopy for authentication purposes. Some studies on distinguishing confused species from the same genus and on exploring distributions of chemicals in tissues of CHMs by fluorescence microscopy have been reported; however, no systematic investigations on fluorescent characteristics of powdered CHMs have been reported. Here, 46 samples of 16 CHMs were investigated. Specifically, the mechanical tissues including stone cells and fibers, the conducting tissues including three types of vessels, and ergastic substances including crystals of calcium oxalate and secretions, in various powdered CHMs were investigated by both light microscope and fluorescence microscope. The results showed many microscopic features emit fluorescence that makes them easily observed, even against complex backgrounds. Under the fluorescence microscope, different microscopic features from the same powdered CHM or some same features from different powdered CHMs emitted the different fluorescence, making this information very helpful for the authentication of CHMs in powder form. Moreover, secretions with unique chemical profiles from different powdered CHMs showed different fluorescent characteristics. Hence, fluorescence microscopy could be a useful additional method for the authentication of powdered CHMs if the fluorescent characteristics of specific CHMs are known.

The influence of freezer storage of urine samples on the BONN-Risk-Index for calcium oxalate crystallization.

This study was performed to quantify the effect of a 1-week freezer storage of urine on its calcium oxalate crystallization risk. Calcium oxalate is the most common urinary stone material observed in urolithiasis patients in western and affluent countries. The BONN-Risk-Index of calcium oxalate crystallization risk in human urine is determined from a crystallization experiment performed on untreated native urine samples. We tested the influence of a 1-week freezing on the BONN-Risk-Index value as well as the effect of the sample freezing on the urinary osmolality. In vitro crystallization experiments in 49 native urine samples from stone-forming and non-stone forming individuals were performed in order to determine their calcium oxalate crystallization risk according to the BONN-Risk-Index approach. Comparison of the results derived from original sample investigations with those obtained from the thawed aliquots by statistical evaluation shows that i) no significant deviation from linearity between both results exists and ii) both results are identical by statistical means. This is valid for both, the BONN-Risk-Index and the osmolality data. The differences in the BONN-Risk-Index results of both procedures of BONN-Risk-Index determination, however, exceed the clinically acceptable difference. Thus, determination of the urinary calcium oxalate crystallization risk from thawed urine samples cannot be recommended.

An unidentified macromolecular inhibitory constituent of calcium oxalate crystal growth in human urine.

We have detected and isolated a macromolecular constituent in normal human urine possessing calcium crystal growth inhibitory activity. The purification procedure consisted of two anion exchange chromatographies and one affinity chromatography. The crystal growth inhibitor was found to be heterogeneous in net charge as well as in size. It has not been identified. It is not an uronic acid-containing glycosaminoglycan, hitherto presumed to be responsible for the inhibitory activity. Whether an urinary fragment of inter-alpha-trypsin inhibitor is responsible has yet to be resolved.