Carboxymethylated Desmodium styracifolium polysaccharide reduces the risk of calcium oxalate kidney stone formation by inhibiting crystal adhesion and promoting crystal endocytosis.

The inhibition of cell surface crystal adhesion and an appropriate increase in crystal endocytosis contribute to the inhibition of kidney stone formation. In this study, we investigated the effects of different degrees of carboxymethylation on these processes. An injury model was established by treating human renal proximal tubular epithelial (HK-2) cells with 98.3 ± 8.1 nm calcium oxalate dihydrate (nanoCOD) crystals. The HK-2 cells were protected with carboxy (-COOH) Desmodium styracifolium polysaccharides at 1.17% (DSP0), 7.45% (CDSP1), 12.2% (CDSP2), and 17.7% (CDSP3). Changes in biochemical indexes and effects on nanoCOD adhesion and endocytosis were detected. The protection of HK-2 cells from nanoCOD-induced oxidative damage by carboxymethylated Desmodium styracifolium polysaccharides (CDSPs) is closely related to the protection of subcellular organelles, such as mitochondria. CDSPs can reduce crystal adhesion on the cell surface and maintain appropriate crystal endocytosis, thereby reducing the risk of kidney stone formation. CDSP2 with moderate -COOH content showed the strongest protective activity among the CDSPs.

Antioxidant Sulfated Polysaccharide from Edible Red Seaweed Gracilaria birdiae is an Inhibitor of Calcium Oxalate Crystal Formation.

The genus Gracilaria synthesizes sulfated polysaccharides (SPs). Many of these SPs, including those synthesized by the edible seaweed Gracilaria birdiae, have not yet been adequately investigated for their use as potential pharmaceutical compounds. Previous studies have demonstrated the immunomodulatory effects of sulfated galactans from G. birdiae. In this study, a galactan (GB) was extracted from G. birdiae and evaluated by cell proliferation and antioxidant tests. GB showed no radical hydroxyl (OH) and superoxide (O2-) scavenging ability. However, GB was able to donate electrons in two further different assays and presented iron- and copper-chelating activity. Urolithiasis affects approximately 10% of the world's population and is strongly associated with calcium oxalate (CaOx) crystals. No efficient compound is currently available for the treatment of this disease. GB appeared to interact with and stabilize calcium oxalate dihydrate crystals, leading to the modification of their morphology, size, and surface charge. These crystals then acquired the same characteristics as those found in healthy individuals. In addition, GB showed no cytotoxic effect against human kidney cells (HEK-293). Taken together, our current findings highlight the potential application of GB as an antiurolithic agent.

Modulation of calcium oxalate dihydrate growth by phosphorylated osteopontin peptides.

Osteopontin (OPN) is a significant component of kidney stone matrix and a key modulator of stone formation. Here, we investigated the effects of different phosphorylated states of a synthesized peptide of OPN (the ASARM peptide; acidic, serine- and aspartate-rich motif) on calcium oxalate dihydrate (COD) crystals, a major mineral phase of kidney stones. In vitro, phosphorylated OPN-ASARM peptides strongly inhibited COD crystal growth in solution as compared to the nonphosphorylated state, with increasing inhibitory potency correlating with the degree of peptide phosphorylation. Scanning electron microscopy revealed that the inhibition from the phosphopeptides resulted in distinctive, rosette-like crystal aggregates called spherulites. The OPN-ASARM peptides preferentially bound and specifically inhibited the {1 1 0} crystallographic faces of COD, as identified by combining atomic force microscopy and computational simulation approaches. These {1 1 0} surfaces of COD have high lattice calcium occupancy (exposure), providing preferential binding sites for the highly acidic peptides; binding and inhibition by OPN-ASARM peptides at the {1 1 0} faces led to crystal aggregation and intergrowth. The crystal spherulite formations obtained in vitro when using the most phosphorylated form of OPN-ASARM peptide at a high concentration, resembled crystal morphologies observed in vivo in a rat model of urolithiasis, in which crystal deposits in the kidney contain abundant OPN as revealed by immunogold labeling. A mechanistic model for spherulite formation is proposed based on the symmetry and crystallographic structure of COD, where the phosphate groups of OPN-ASARM bind to calcium atoms at [1 1 1] step risers on the COD {1 1 0} surface, inducing the periodic emergence of new COD crystals to form spherulites.

Non-Invasive characterisation of renal stones using dual energy CT: A method to differentiate calcium stones.

BACKGROUND: Non-invasive DECT based characterization of renal stones using their effective atomic number (Zeff) and the electron density (ρe) in patients. AIM: This paper aims to develop a method for in-vivo characterization of renal stone. Differentiation of renal stones in-vivo especially sub types of calcium stones have very important advantage for better judgement of treatment modality. MATERIALS AND METHODS: 50 extracted renal stones were scanned ex-vivo using dual energy CT scanner. A method was developed to characterize these renal stones using effective atomic number and electron density obtained from dual energy CT data. The method and formulation developed in ex-vivo experiments was applied in in-vivo study of 50 randomly selected patients of renal stones who underwent dual energy CT scan. RESULTS: The developed method was able to characterize Calcium Oxalate Monohydrate (COM) and the combination of COM and Calcium Oxalate Dihydrate (COD) stones non-invasively in patients with a sensitivity of 81% and 83%respectively. The method was also capable of differentiating Uric, Cystine and mixed stones with the sensitivity of 100, 100 and 85.71% respectively. CONCLUSION: The developed dual energy CT based method was capable of differentiating sub types of calcium stones which is not differentiable on single energy or dual energy CT images.

Endoscopic identification of urinary stone composition: A study of South Eastern Group for Urolithiasis Research (SEGUR 2). / Identificación endoscópica de la composición de los cálculos urinarios: un estudio del Southeastern Group for Lithiasis Research (SEGUR 2).

INTRODUCTION AND OBJECTIVES: To assess the surgeon's ability to evaluate the composition of the stone by observation of endoscopic images. MATERIALS AND METHODS: A series of 20 video clips of endoscopic treatments of urinary stones of which was also available the result of infrared spectroscopy was uploaded to a YouTube site accessible only to members of the South Eastern Group for Urolithiasis Research (SEGUR) who were asked to identify the composition of the stones. RESULTS: A total of 32 clinicians from 9 different countries participated in the study. The average number of correct detections of participants was 7.81 ± 2.68 (range 1-12). Overall accuracy was 39% (250 out of 640 predictions). Calcium oxalate dihydrate stones have been correctly detected in 69.8%, calcium oxalate monohydrate in 41.8%, uric acid in 33.3%, calcium oxalate/uric acid in 34.3% and cystine in 78.1%. Precision rates for struvite (15.6%), calcium phosphate (0%) and mixed calcium oxalate/calcium phosphate (9.3%) were quite low. CONCLUSIONS: Observation of the stone during the endoscopic procedure was not reliable to identify the composition of most stones although it gave some information allowing to identify with a good sensitivity calcium oxalate dihydrate and cystine stones. Nevertheless, photo or video reporting of the intact stone and its internal structure could should be encouraged to implement results of still mandatory post-operative stone analysis. Endourologists should improve their ability of visual identification of the different types of stones.

The relationship between calcium oxalate lithiasis and chronic proinflammatory intestinal dysbiosis pattern: a prospective study.

The objective is to establish whether a pattern of intestinal dysbiosis exists in calcium oxalate (CaOx) lithiasis and, if so, to identify its characteristics and explore whether there are differences in the pattern between CaOx dihydrate (COD) and monohydrate (COM) lithiasis. With this aim 24 patients diagnosed with CaOx lithiasis by means of optical microscopy and spectrometry were prospectively recruited. Faecal analysis was carried out by means of RT-PCR 16S rRNA assay and agar plate culture according to the methodology proposed by the Institute of Microecology (Herborn, Germany). The total number of bacteria was depleted due to COD lithiasis (p = 0.036). The mean values of immunoregulating microbiota were normal, but the percentage of normal values was lower in the COD group (30%) than in the COM group (69.2%) (p = 0.062). The total mean values of protective microbiota were normal in both groups. There was a large decrease in the mean values of the muconutritive microbiota Akkermansia muciniphila and Faecalibacterium prausnitzii, the most intense decline being observed in the COD group (p = 0.019). Levels of proteolytic microbiota were elevated in both groups, without differences between them. We conclude that patients with CaOx lithiasis have a chronic pro-inflammatory intestinal dysbiosis pattern characterised by a reduction in the total number of bacteria, a reduction in immunoregulating microbiota and a large reduction in muconutritive microbiota that is significantly more intense in COD lithiasis than in COM lithiasis.

Potentially Pathogenic Calcium Oxalate Dihydrate and Titanium Dioxide Crystals in the Alzheimer's Disease Entorhinal Cortex.

Knowing that Alzheimer's disease (AD) nucleates in the entorhinal cortex (EC), samples of 12 EC specimens were probed for crystals by a protocol detecting fewer than 1/5000th of those present. Of the 61 crystals found, 31 were expected and 30 were novel. Twenty-one crystals of iron oxides and 10 atherosclerosis-associated calcium pyrophosphate dihydrate crystals were expected and found. The 30 unexpected crystals were NLRP3-inflammasome activating calcium oxalate dihydrate (12) and titanium dioxide (18). Their unusual distribution raises the possibility that some were of AD origination sites.

The effect of hydrodynamic and thermodynamic factors and the addition of citric acid on the precipitation of calcium oxalate dihydrate.

This paper reports on the investigation of experimental conditions relevant for spontaneous precipitation of significant amount of pure calcium oxalate dihydrate (COD). For this purpose, the hydrodynamic and thermodynamic parameters, such as mode of agitation, temperature, supersaturation and concentration of additives (citrate ions), have been studied. The results show that in the model systems, without the citrate addition and applied mechanical stirring, calcium oxalate monohydrate (COM) was observed as dominant modification after 20 min of aging, while the magnetic stirring resulted in a formation of a mixture of COM and calcium oxalate trihydrate (COT), regardless of the temperature applied. In the mechanically stirred systems, the addition of citrate ions in the range of concentrations, 0.001 mol dm-3 < c i (Na3C6H5O7) < 0.012 mol dm-3, caused the formation of COM and COD mixture at all temperatures. At the same conditions and in the magnetically stirred systems formation of COD, in a mixture with COT or COM, has been observed. The highest COD content in the mechanically stirred system was obtained at 45 °C and c i (Na3C6H5O7) = 0.001 mol dm-3 (w = 89.5%), while in the magnetically stirred system almost pure COD was obtained at 37 °C and c i (Na3C6H5O7) = 0.008 mol dm-3 (w = 96.5%).

Effect of Blumea balsamifera extract on the phase and morphology of calcium oxalate crystals.

OBJECTIVE: Calcium oxalate crystals are found in majority of kidney stones with calcium oxalate monohydrate (COM) as one of the primary types of kidney stones. Various methods of treatment exist, including herbal treatment in the Philippines that uses the medicinal herb Blumea balsamifera (B. balsamifera). METHODS: The effect of B. balsamifera extract on the morphology of calcium oxalate crystals was studied by light microscopy, image analysis, powder X-ray diffraction and scanning electron microscopy. RESULTS: The extract decreased the crystal size by 5.22%-82.62% depending on the degree of supersaturation. Through analysis of the projected area of the crystals, the extract was found to shift the phase of the crystals from COM to calcium oxalate dihydrate (COD). This shift in phase is significant with a COM to COD shift of 88.26% at 0.5 mg/mL of extract and 91.53% at 1.0 mg/mL of extract. Scanning election microscopic (SEM) images revealed aggregation of crystals at 0 mg/mL of extract. At 1.0 mg/mL of extract, the scanning electron micrographs showed discernible crystal unit boundaries. CONCLUSION: B. balsamifera extract was observed to have decreased crystal size, shifted crystal phase from COM to COD and prevented the aggregation of calcium oxalate crystals.

A Case of Randall's Plugs Associated to Calcium Oxalate Dihydrate Calculi.

A case of a patient who developed multiple calcium oxalate dihydrate calculi, some of them connected to intratubular calcifications (Randall's plugs), is presented. Randall's plugs were isolated and studied. The mechanism of Randall's plug development is also suggested.

Preparation, characterization, and in vitro cytotoxicity of COM and COD crystals with various sizes.

Calcium oxalate crystals in urine often differ in size and crystal phase between healthy humans and patients with kidney stones. In this work, calcium oxalate monohydrate (COM) and dihydrate (COD) with sizes of about 50 nm, 100 nm, 1 µm, 3 µm, and 10 µm were prepared by varying reactant concentration, reaction temperature, solvent, mixing manner, and stirring speed. These crystals mainly had a smooth surface and no obvious pore structure, except COM-1 µm. In cell culture medium, the zeta potential of crystals became increasingly negative with increasing size, and the absolute value of zeta potential of COD was greater than the same-sized COM. Results of cell viability and PI staining assays showed that the order of injury degree in African green monkey renal epithelial (Vero) cells caused by different sizes of COD was COD-50 nm>COD-100 nm>COD-1 µm>COD-3 µm>COD-10 µm, and that of different sizes of COM was COM-1 µm>COM-50~COM-100 nm>COM-3 µm>COM-10 µm. COM-1 µm presented the highest cytotoxicity in Vero cells, which was associated with its rougher surface, larger specific surface area (SBET), and larger pore volume. Overall, these findings indicated that the physical properties of crystals play an important role in their cytotoxicity.

Characterisation of the turbid particles in the extraction of sugar beet pectins.

This paper was aimed at characterising the insoluble substances (IS) responsible for the turbidity of the extract and impurity of the resulting pectins. Results showed that the IS caused a significant increase in the turbidity of the extract. The IS had bi-pyramidal shapes under the SEM observation. The observed XRD peaks for the IS were similar to those of calcium oxalate dihydrate (COD). Moreover, the IS consisted of mainly oxalate and calcium. Results from the present study indicate the IS is COD. The influence of the IS on the purity of pectin was also studied. The presence of the IS in the pectins indicated the IS can precipitate with pectins during the alcohol precipitation, thereby contaminating the resulting pectins.

Interaction between submicron COD crystals and renal epithelial cells.

OBJECTIVES: This study aims to investigate the adhesion characteristics between submicron calcium oxalate dihydrate (COD) with a size of 150 ± 50 nm and African green monkey kidney epithelial cells (Vero cells) before and after damage, and to discuss the mechanism of kidney stone formation. METHODS: Vero cells were oxidatively injured by hydrogen peroxide to establish a model of injured cells. Scanning electron microscopy was used to observe Vero-COD adhesion. Inductively coupled plasma emission spectrometry was used to quantitatively measure the amount of adhered COD microcrystals. Nanoparticle size analyzer and laser scanning confocal microscopy were performed to measure the change in the zeta potential on the Vero cell surface and the change in osteopontin expression during the adhesion process, respectively. The level of cell injury was evaluated by measuring the changes in malonaldehyde content, and cell viability during the adhesion process. RESULTS: The adhesion capacity of Vero cells in the injury group to COD microcrystals was obviously stronger than that of Vero cells in the control group. After adhesion to COD, cell viability dropped, both malonaldehyde content and cell surface zeta potential increased, and the fluorescence intensity of osteopontin decreased because the osteopontin molecules were successfully covered by COD. Submicron COD further damaged the cells during the adhesion process, especially for Vero cells in the control group, leading to an elevated amount of attached microcrystals. CONCLUSION: Submicron COD can further damage injured Vero cells during the adhesion process. The amount of attached microcrystals is proportional to the degree of cell damage. The increased amount of microcrystals that adhered to the injured epithelial cells plays an important role in the formation of early-stage kidney stones.