CaOx Stone Research Articles

Current Dietary and Medical Prevention of Renal Calcium Oxalate Stones.

Kidney stones refer to abnormal crystal formation that occurs in the kidney. Among a variety of components of kidney stones, calcium oxalate (CaOx) is the most common type. Despite many efforts to investigate the pathogenesis of CaOx stones, the pathogenesis remains an issue of debate. With high occurrence and recurrence, individuals with stone formation are prone to frequently consult a doctor and to be hospitalized, and the treatment of kidney stones poses a heavy burden on the patients. Concerns should be focused not only on treatment but also on prevention. Herein, we reviewed the studies on prevention methods of CaOx stones through diet, lifestyle, and medication extending until the current time frame. As hyperoxaluria is the most common metabolic disorder among CaOx stone formations, we also included several studies on the treatment and prevention of hyperoxaluria. Our objective was to outline the effective methods to prevent renal CaOx stone formation.

431 Defective uromodulin polymerization and peptide excretion in a natural canine model of kidney stones

OBJECTIVES/GOALS: Using a natural canine model of kidney stone disease, we previously identified a pathogenic variant in the uromodulin gene (UMOD) that imparts a dramatic risk for calcium oxalate (CaOx) stones. This study was designed to characterize the effects of the pathogenic variant on uromodulin processing, specifically polymerization and peptide excretion. METHODS/STUDY POPULATION: Uromodulin polymerization status and peptides were measured in random urine samples from CaOx stone-forming dogs with the pathogenic UMOD variant and breed-, sex-, and age-matched healthy control dogs. Polymerization status was determined using an ultracentrifugation protocol and Western blotting in 6 CaOx cases and 3 controls; relative abundance of the polymerizing and nonpolymerizing forms was evaluated. Uromodulin peptide abundances were measured by LC-MS/MS with 4 dogs per group; results were summed to determine total uromodulin peptide excretion for each dog, and individual peptide abundances were calculated as a percentage of the total. Polymerization status and peptides were compared between groups. RESULTS/ANTICIPATED RESULTS: Dogs with the pathogenic UMOD variant had abnormalities in both uromodulin polymerization and peptide processing. The polymerization data showed that the polymerizing form of uromodulin was abundant in all healthy controls but absent or severely reduced in most dogs with the variant. In contrast, nonpolymerizing uromodulin was detected in all dogs with no observed difference between those with and without the variant. The peptidomics data showed that stone-forming dogs with the pathogenic UMOD variant lacked a peptide cleavage site, resulting in the loss of two common peptides that terminate at that site and the presence of longer peptides that span the site. DISCUSSION/SIGNIFICANCE: These findings implicate uromodulin polymerization and peptide processing defects in kidney stone risk. Future studies will define the mechanisms through which these defects affect stone formation, ultimately informing development of novel preventative therapies.

Identification and validation of the biomarkers related to ferroptosis in calcium oxalate nephrolithiasis.

Ferroptosis is a specific type of programmed cell death characterized by iron-dependent lipid peroxidation. Understanding the involvement of ferroptosis in calcium oxalate (CaOx) stone formation may reveal potential targets for this condition. The publicly available dataset GSE73680 was used to identify 61 differentially expressed ferroptosis-related genes (DEFERGs) between normal kidney tissues and Randall's plaques (RPs) from patients with nephrolithiasis through employing weighted gene co-expression network analysis (WGCNA). The findings were validated through in vitro and in vivo experiments using CaOx nephrolithiasis rat models induced by 1% ethylene glycol administration and HK-2 cell models treated with 1 mM oxalate. Through WGCNA and the machine learning algorithm, we identified LAMP2 and MDM4 as the hub DEFERGs. Subsequently, nephrolithiasis samples were classified into cluster 1 and cluster 2 based on the expression of the hub DEFERGs. Validation experiments demonstrated decreased expression of LAMP2 and MDM4 in CaOx nephrolithiasis animal models and cells. Treatment with ferrostatin-1 (Fer-1), a ferroptosis inhibitor, partially reversed oxidative stress and lipid peroxidation in CaOx nephrolithiasis models. Moreover, Fer-1 also reversed the expression changes of LAMP2 and MDM4 in CaOx nephrolithiasis models. Our findings suggest that ferroptosis may be involved in the formation of CaOx kidney stones through the regulation of LAMP2 and MDM4.

The modulatory effects of large and small extracellular vesicles from normal human urine on calcium oxalate crystallization, growth, aggregation, adhesion on renal cells, and invasion through extracellular matrix: An in vitro study

Urinary extracellular vesicles (uEVs) play important roles in physiologic condition and various renal/urological disorders. However, their roles in kidney stone disease remain unclear. This study aimed to examine modulatory effects of large and small uEVs derived from normal human urine on calcium oxalate (CaOx) crystals (the main component in kidney stones). After isolation, large uEVs, small uEVs and total urinary proteins (TUPs) with equal (protein equivalent) concentration were added into various crystal assays to compare with the control (without uEVs or TUPs). TUPs strongly inhibited CaOx crystallization, growth, aggregation and crystal-cell adhesion. Large uEVs had lesser degree of inhibition against crystallization, growth and crystal-cell adhesion, and comparable degree of aggregation inhibition compared with TUPs. Small uEVs had comparable inhibitory effects as of TUPs for all these crystal assays. However, TUPs and large uEVs slightly promoted CaOx invasion through extracellular matrix, whereas small uEVs did not affect this. Matching of the proteins reported in six uEVs datasets with those in the kidney stone modulator (StoneMod) database revealed that uEVs contained 18 known CaOx stone modulators (mainly inhibitors). These findings suggest that uEVs derived from normal human urine serve as CaOx stone inhibitors to prevent healthy individuals from kidney stone formation.

Establishment and application of a nomogram diagram for predicting calcium oxalate stones in patients with urinary tract stones

This retrospective study aims to examine the correlation between calcium oxalate (CaOx) stones and common clinical tests, as well as urine ionic composition. Additionally, we aim to develop and implement a personalized model to assess the accuracy and feasibility of using charts to predict calcium oxalate stones in patients with urinary tract stones. A retrospective analysis was conducted on data from 960 patients who underwent surgery for urinary stones at the First Affiliated Hospital of Soochow University from January 1, 2010, to December 31, 2022. Among these patients, 447 were selected for further analysis based on screening criteria. Multivariate logistic regression analysis was then performed to identify the best predictive features for calcium oxalate stones from the clinical data of the selected patients. A prediction model was developed using these features and presented in the form of a nomogram graph. The performance of the prediction model was assessed using the C-index, calibration curve, and decision curve, which evaluated its discriminative power, calibration, and clinical utility, respectively. The nomogram diagram prediction model developed in this study is effective in predicting calcium oxalate stones which is helpful in screening and early identification of high-risk patients with calcium oxalate urinary tract stones, and may be a guide for urologists in making clinical treatment decisions.

Dysregulated palmitic acid metabolism promotes the formation of renal calcium-oxalate stones through ferroptosis induced by polyunsaturated fatty acids/phosphatidic acid

The pathogenesis of renal calcium-oxalate (CaOx) stones is complex and influenced by various metabolic factors. In parallel, palmitic acid (PA) has been identified as an upregulated lipid metabolite in the urine and serum of patients with renal CaOx stones via untargeted metabolomics. Thus, this study aimed to mechanistically assess whether PA is involved in stone formation. Lipidomics analysis of PA-treated renal tubular epithelial cells compared with the control samples revealed that α-linoleic acid and α-linolenic acid were desaturated and elongated, resulting in the formation of downstream polyunsaturated fatty acids (PUFAs). In correlation, the levels of fatty acid desaturase 1 and 2 (FADS1 and FADS2) and peroxisome proliferator-activated receptor α (PPARα) in these cells treated with PA were increased relative to the control levels, suggesting that PA-induced upregulation of PPARα, which in turn upregulated these two enzymes, forming the observed PUFAs. Lipid peroxidation occurred in these downstream PUFAs under oxidative stress and Fenton Reaction. Furthermore, transcriptomics analysis revealed significant changes in the expression levels of ferroptosis-related genes in PA-treated renal tubular epithelial cells, induced by PUFA peroxides. In addition, phosphatidyl ethanolamine binding protein 1 (PEBP1) formed a complex with 15-lipoxygenase (15-LO) to exacerbate PUFA peroxidation under protein kinase C ζ (PKC ζ) phosphorylation, and PKC ζ was activated by phosphatidic acid derived from PA. In conclusion, this study found that the formation of renal CaOx stones is promoted by ferroptosis of renal tubular epithelial cells resulting from PA-induced dysregulation of PUFA and phosphatidic acid metabolism, and PA can promote the renal adhesion and deposition of CaOx crystals by injuring renal tubular epithelial cells, consequently upregulating adhesion molecules. Accordingly, this study provides a new theoretical basis for understanding the correlation between fatty acid metabolism and the formation of renal CaOx stones, offering potential targets for clinical applications.

Contrasting Response of Urine Stone Risk to Medical Treatment in Calcium Oxalate versus Calcium Phosphate Stone Formers.

Randomized controlled trials have shown that both thiazide diuretics and potassium citrate (K-Cit) can prevent calcium stone recurrence, but most participants formed calcium oxalate stones (CaOx). While thiazides are expected to lower stone risk for calcium phosphate (CaP) stones, the effect of K-Cit on risk for CaP stones is unclear. To study the impact of common calcium stones treatments, we analyzed the 24-hour urines of CaOx and CaP stone formers (SF) by 4 treatment types: Lifestyle, K-Cit, Thiazide, or Both medications. Patients treated with thiazides reduced urine calcium in both CaOx (M = -74.4, SD = 94.6 mg/day) and CaP SF (M = -102, SD = 99.7 mg/day), while those on K-Cit had no change in urine calcium. Among CaOx SF, urine citrate rose in patients administered K-Cit with or without thiazide, but citrate did not rise significantly in CaP SF. Urine pH rose in all CaOx SF, but among CaP SF only rose in patients receiving K-Cit. CaOx supersaturation (SS) decreased in all patients who received Thiazide, and decreased among CaOx SF treated with K-Cit. CaP SS decreased in both CaOx SF (M = -0.46, SD = 0.86) and CaP SF (M = -0.76, SD = 0.85) treated with Thiazide except CaOx SF who received Both. Patients treated with K-Cit alone increased CaP SS in CaOx SF (M = 0.25, SD = 0.79). Patients treated with Thiazide lowered urine calcium and SS in both stone groups. Patients treated with K-Cit had no significant changes in urine calcium and had a decrease in CaOx SS in CaOx SF. The study raises questions about the best preventive treatment for patients with CaP stones and suggests that K-Cit may not confer the same benefits on CaP SF as it does on CaOx SF.

CCL7 and olfactory transduction pathway activation play an important role in the formation of CaOx and CaP kidney stones.

Background: The deposition of calcium oxalate (CaOx) and calcium phosphate (CaP) is the most common cause of kidney stone disease (KSD). Whether KSDs caused by CaOx and CaP have common genetic targets or signaling pathways remained unclear. Methods: The present study utilized public data GSE73680 to analyze differentially expressed genes between CaOx or CaP tissues and normal tissues, respectively. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of co-DEGs were performed. The protein-protein interaction (PPI) network was constructed to identify hub genes, and the top hub gene was selected for gene set enrichment analysis (GSEA). Finally, real-time PCR of patients' urine was performed to validate the bioinformatic results. Results: In total, 155 significantly co-upregulated DEGs and 64 co-downregulated DEGs were obtained from the datasets. The Gene Ontology analysis showed that DEGs were significantly enriched in chemical stimulus in sensory perception, detection of chemical stimulus in sensory perception of smell, and olfactory receptor activity. The KEGG analysis showed that the olfactory transduction pathway was significantly enriched. According to protein-protein interaction, 10 genes were identified as the hub genes, and CCL7 was the top hub gene. The olfactory transduction, maturity-onset diabetes of the young, linoleic acid metabolism, and fat digestion and absorption were significantly enriched in the high-CCL7 subgroup by GSEA. In total, 9 patients who had primarily CaOx mixed with some CaP stones and 9 healthy subjects were enrolled. The RT-PCR results showed that CCL7 level in the stone group was significantly higher than that in the control group (p < 0.05). For the olfactory transduction pathway, the expression of OR10A5, OR9A2, and OR1L3 was significantly upregulated in the stone group compared with the control group (p < 0.05). Conclusion: CCL7 may play a key role in the development of both CaOx and CaP, and this process may depend on olfactory transduction pathway activation.

Resveratrol inhibits calcium oxalate crystal growth, reduces adhesion to renal cells and induces crystal internalization into the cells, but promotes crystal aggregation

Resveratrol is a natural phenolic compound that belongs to stilbenoid group found in diverse plants. Health benefits and therapeutic potentials of resveratrol have been widely recognized in various diseases. In kidney stone disease, it can alleviate oxalate-induced hyperproduction of free radicals in renal epithelial cells. Nevertheless, its direct effects on calcium oxalate (CaOx) crystal, which is the major stone component, remained unclear. This study therefore addressed the direct effects of resveratrol (at 1, 10 or 100 μM) on each step of CaOx kidney stone formation. The results revealed that resveratrol had no significant effects on CaOx crystallization. However, resveratrol significantly decreased CaOx crystal growth and adhesion to renal epithelial cells at all concentrations, and induced crystal internalization into the cells (a process related to crystal degradation by endolysosomes) in a concentration-dependent manner. On the other hand, resveratrol promoted crystal aggregation. These data indicate that resveratrol serves as a dual modulator on CaOx stone formation. While it inhibits CaOx stone development by reducing crystal growth and adhesion to renal cells and by inducing crystal internalization into the cells, resveratrol promotes crystal aggregation, which is one of the mechanisms leading to kidney stone formation.

Antioxidant Activity of Auricularia auricula Polysaccharides with Different Molecular Weights and Cytotoxicity Difference of Polysaccharides Regulated CaOx to HK-2 Cells.

This study aimed to investigate the growth of calcium oxalate (CaOx) crystals regulated by Auricularia auricular polysaccharides (AAPs) with different viscosity-average molecular weights (Mv), the toxicity of AAP-regulated CaOx crystals toward HK-2 cells, and the prevention and treatment capabilities of AAPs for CaOx stones. The scavenging capability and reducing capacity of four kinds of AAPs (Mv of 31.52, 11.82, 5.86, and 3.34 kDa) on hydroxyl, ABTS, and DPPH free radicals and their capability to chelate divalent iron ions were detected. AAP-regulated CaOx crystals were evaluated by using zeta potential, thermogravimetric analysis, X-ray diffraction, and scanning electron microscopy. The cytotoxicity of AAP-regulated crystals was evaluated through examination of cell viability, cell death, malondialdehyde (MDA) content, and cell surface hyaluronic acid (HA) expression. The in vitro antioxidant activities of the four AAPs were observed in the following order: AAP0 < AAP1 < AAP2 < AAP3. Thus, AAP3, which had the smallest Mv, had the strongest antioxidant activity. AAPs can inhibit the growth of CaOx monohydrate (COM), induce the formation of CaOx dihydrate (COD), and reduce the degree of crystal aggregation, with AAP3 exhibiting the strongest capability. Cell experiments showed the lowest cytotoxicity in AAP3-regulated CaOx crystals, along with the lowest MDA content, HA expression, and cell mortality. In addition, COD presented less cytotoxicity than COM. Meanwhile, the cytotoxicity of blunt crystals was less than that of sharp crystals. AAPs, particularly AAP3, showed an excellent antioxidative capability in vitro, and AAP3-regulated CaOx crystals presented minimal cytotoxicity.

Diagnostic value of urinary Tamm-Horsfall protein and 24 h urine osmolality for recurrent calcium oxalate stones of the upper urinary tract: Cross-sectional study.

We aimed to investigate the diagnostic value of urinary Tamm-Horsfall protein (THP) and 24 h urine osmolality for recurrent calcium oxalate (CaOx) stones. Clinical data of 120 patients with upper urinary tract stones admitted to our hospital between January 2020 and January 2022 were retrospectively reviewed. Patients were divided into recurrence (53 patients) and non-recurrence (67 patients) groups based on postoperative stone recurrence. Meanwhile, 50 healthy patients were selected as the control group. Urinary THP levels, 24 h urine osmolality, and biochemical indices were compared between the three groups; their diagnostic values for stone recurrence were evaluated using receiver operating characteristic (ROC) curves. Urinary THP, 24 h urine osmolality, and biochemical indices were significantly higher in the recurrence group than in the non-recurrence and control groups (P < 0.05). The 24 h urine osmolality was positively correlated with urinary oxalic acid and calcium excretion. ROC curve analysis showed that optimal cutoff values of urinary THP and 24 h urine osmolality for the diagnosis of stone development were ≥27.01 mg/L and ≥577.69 mOsm/(kg H2O), respectively. Furthermore, these indices combined significantly improved the accuracy of diagnosis. Urinary THP and 24 h urine osmolality were higher in patients with recurrent CaOx stones. Detection of both parameters combined can accurately diagnose stone recurrence.

The first epidemiology of urolithiasis in Northern Vietnam: Urinary stone composition, age, gender, season, and clinical features study.

The aim of this study is to analyze the compositions of urinary stones and investigate their distributions in different ages, genders, seasons, and clinical features of Northern Vietnamese patients. A total of 231 patients with urinary stones from Northern Vietnam were collected and analyzed composition from 1/2021-12/2022. For all patients, age, sex, stone location, stone side, urine pH, and hospitalized date (month) were collected. Kidney stones are more frequently found in men than women with the male: female urinary stones ratio in this study being 1.96:1. The highest stone prevalence appeared between 60 and 69 years old. The most common stone composition was calcium oxalate, followed by calcium phosphate, uric acid, struvite, and cysteine. Mix stones of CaOx and CaP were more prevalent than pure stones. Males submitted more CaOx, CaP, and UA stones, whereas females were susceptible to infectious stones. Stones were more frequently found on the left side of the upper urinary tract (51.9%) than on the right side (27.3%) and lower urinary tract (7.8%). Cultural tendency leads to a smaller number of stones during the Lunar new year (February), and Ghost month (August).

The genetics of urinary microbiome, an exploration of the trigger in calcium oxalate stone.

Background: Kidney stone disease is a global disease; however, it has not been totally understood. Calcium oxalate (CaOx) stone is the dominant type of kidney stone, and the potential factors involved in its formation are yet to be explored. Clinically, we found that the CaOx stones in patients were mainly unilateral; therefore, systemic factors cannot explain them, although some local factors must be involved. Urinary microbiota is involved in stone formation. Therefore, this study aimed to explore the association between the urinary microbiota and CaOx stones and provide insight into the medical treatment and prevention of CaOx stones. Methods: Sixteen pelvic urine samples were collected from the stone and non-stone sides of patients with unilateral CaOx stones following strict criteria. The 16S rRNA gene sequencing was performed on each pair of pelvic urine samples at the species level. Many bioinformatic analyses were conducted to explore the potential factors affecting CaOx stone formation. Results: Although no statistically significant difference was found between the overall microbiota of the pelvis urine from the two sides. Many biologically distinct taxa were observed, including many bacteria found in previous studies, like Proteobacteria, Actinobacteria, Firmicute and Enterobacter cloacae and so on. What's more, despite these common bacteria, our current study added to these bacterial communities with additional identification of Deinococcus-Thermus, Coriobacteriia, Porphyromonas and Ralstonia. To predict the functions of these microbiota, Kyoto Encyclopedia for Genes and Genomes and MetaCyc analysis were conducted and immunometabolism might be an important pathway. Moreover, a random forest predictor was constructed to distinguish the stone side from the non-stone side, with an accuracy of 62.5%. Conclusion: Our research profiled the microbiome in the pelvis urine from both the stone and non-stone sides of patients with unilateral CaOx stones, provided insight into the dominant role of urinary dysbiosis in CaOx stones formation. Furthermore, this study also predicted the potential crosstalk among urinary microbiota, immunometabolism, and CaOx stones.

Nontargeted Urine Metabolomics with Electrospray Ionization-Mass Spectrometry Reveals the Major Metabolic Characterization between Adult Males and Females Involved in Calcium Oxalate Urolithiasis

The incidence of calcium oxalate (CaOx) urolithiasis between men and women is considerably different. However, the full-scale metabolic changes between genders with CaOx urolithiasis have not been reported. The metabolic profiles in 112 urine samples obtained from 61 males and 51 females diagnosed with CaOx stones were characterized using electrospray ionization-mass spectrometry (ESI-MS). Univariate, multivariate, and bioinformatic analyses were employed to reveal the differential metabolites and perturbed metabolic pathways between genders. In total, 30 dysregulated metabolites were identified as differential metabolites between males and females with CaOx urolithiasis. The primary metabolic pathways were caffeine metabolism, metabolic pathways, and biosynthesis of amino acids. Furthermore, 10 potential metabolites which were mostly contributed to separating the males and female were selected using feature weight calculation. These compounds included creatinine, 3-methylhistidine, n-acetylneuraminic acid, decanoyl-l-carnitine, cortisone, nicotine, 1-methyluric acid, androsterone sulfate, citramalic acid, and 1,7-dimethylxanthine. Finally, a subset of the differential metabolites was identified by receiver operating characteristic (ROC) curve analysis, including creatinine, 3-methylhistidine, n-acetylneuraminic acid, decanoyl-l-carnitine, and 1-methyluric acid (area under the curve = 0.924, 0.844, 0.846, 0.814, 0.814). These metabolites had higher risks in the CaOx stone formation of males than females. The study showed that there were a group of differential metabolites between the males and females with CaOx stones. A subset of these compounds was more closely and positively related to renal stone formation in males than females. Moreover, these metabolites may be the cause of the differences in the incidence of CaOx urolithiasis between genders.

A spatially anchored transcriptomic atlas of the human kidney papilla identifies significant immune injury in patients with stone disease

Kidney stone disease causes significant morbidity and increases health care utilization. In this work, we decipher the cellular and molecular niche of the human renal papilla in patients with calcium oxalate (CaOx) stone disease and healthy subjects. In addition to identifying cell types important in papillary physiology, we characterize collecting duct cell subtypes and an undifferentiated epithelial cell type that was more prevalent in stone patients. Despite the focal nature of mineral deposition in nephrolithiasis, we uncover a global injury signature characterized by immune activation, oxidative stress and extracellular matrix remodeling. We also identify the association of MMP7 and MMP9 expression with stone disease and mineral deposition, respectively. MMP7 and MMP9 are significantly increased in the urine of patients with CaOx stone disease, and their levels correlate with disease activity. Our results define the spatial molecular landscape and specific pathways contributing to stone-mediated injury in the human papilla and identify associated urinary biomarkers.

Inhibition of AT1R/IP3/IP3R-mediated Ca2+ release protects against calcium oxalate crystals-induced renal oxidative stress

Calcium oxalate (CaOx) stones are the most prevalent type of kidney stones. CaOx crystals can stimulate reactive oxygen species (ROS) generation and induce renal oxidative stress to promote stone formation. Intracellular Ca2+ is an important signaling molecule, and an elevation of cytoplasmic Ca2+ levels could trigger oxidative stress. Our previous study has revealed that upregulation of Ang II/AT1R promoted renal oxidative stress during CaOx exposure. IP3/IP3R/Ca2+ signaling pathway activated via Ang II/AT1R is involved in several diseases, but its role in stone formation has not been reported. Herein, we focus on the role of AT1R/IP3/IP3R-mediated Ca2+ release in CaOx crystals-induced oxidative stress and explore whether inhibition of this pathway could alleviate renal oxidative stress. NRK-52E cells were exposed to CaOx crystals pretreated with AT1R inhibitor losartan or IP3R inhibitor 2-APB, and glyoxylic acid monohydrate-induced CaOx stone-forming rats were treated with losartan or 2-APB. The intracellular Ca2+ levels, ROS levels, oxidative stress indexes, and the gene expression of this pathway were detected. Our results showed that CaOx crystals activated AT1R to promote IP3/IP3R-mediated Ca2+ release, leading to increased cytoplasmic Ca2+ levels. The Ca2+ elevation was able to stimulate NOX2 and NOX4 to generate ROS, induce oxidative stress, and upregulate the expression of stone-related proteins. 2-APB and losartan reversed the referred effects, reduced CaOx crystals deposition and alleviated tissue injury in the rat kidneys. In summary, our results indicated that CaOx crystals promoted renal oxidative stress by activating the AT1R/IP3/IP3R/Ca2+ pathway. Inhibition of AT1R/IP3/IP3R-mediated Ca2+ release protected against CaOx crystals-induced renal oxidative stress. 2-APB and losartan might be promising preventive and therapeutic agents for the treatment of kidney stone disease.

Pediatric age-related distribution of calcium oxalate monohydrate and calcium oxalate dihydrate in urinary tract stones: metabolic, gender and ethnic correlates.

Previous studies of pediatric urolithiasis have suggested possible associations between the relative proportions of calcium oxalate dihydrate (COD) and calcium oxalate monohydrate (COM) stones with age, gender, and ethnicity. This study aimed to investigate the composition and distribution of calcium oxalate stones according to these clinical factors and the metabolic correlates of the different subtypes in pediatric stone formers. We retrospectively reviewed the database of all first-time stone formers between 2014 and 2019. Infrared spectrometry was used to determine stone composition. Stones were categorized by their highest relative component and reported as a percentage of occurrences in the cohort as a whole and by patient gender, age (divided into three age groups: 1-5, 6-12, 13-18 years), and ethnicity. Clinical and metabolic correlates were analyzed. Of 2,479 consecutive stones submitted to our chemical stone lab, 220 first-time pediatric stone formers were identified. COD stones were the predominant subtype in the youngest group, and COM stones in the oldest group (OR 0.39, 95% CI:0.18-0.86, p=0.036). In the intermediate-age group (6-12 years), COM stones were more prevalent in Arab boys and COD stones in girls of either ethnicity. COD stones were associated with hypercalciuria (p< 0.0001), and COM stones with hyperoxaluria (p=0.0024). Hypercalciuria and hypocitraturia were the most prevalent abnormalities at ages 1-5 and 13-18, respectively. Analysis of calcium oxalate stone subtypes and their metabolic correlates in stone formers has significant clinical relevance, specifically in children. In the present study, COD stones and hypercalciuria were more common in younger children, and COM stones and hypocitraturia in adolescents. These findings suggest unique, complex interactions driving stone formations in children that may guide a more practical, limited, and cost-effective approach to metabolic evaluations, choice of treatment, and preventive measures, particularly in first-time calcium oxalate pediatric stone-formers.

Urine pH and Citrate as Predictors of Calcium Phosphate Stone Formation.

Urinary parameters including urine pH and citrate are recognized as critical in the pathophysiology of calcium-based stones. The factors contributing to variation in these parameters between calcium oxalate and calcium phosphate stone formers are however not well understood. In this study using readily available laboratory data, we explore these differences to delineate the odds of forming calcium phosphate (CaP) versus calcium oxalate (CaOx) stones. In this single-center retrospective study, we compared serum and urinary parameters between adult CaP stone formers (SF), CaOx SF, and non-stone formers (NSF). Urine pH was higher and urine citrate lower in CaP SF compared to same-sex CaOx SF and NSF. In CaP SF, higher urine pH and lower citrate were independent of markers of dietary acid intake and gastrointestinal alkali absorption, suggesting abnormal renal citrate handling and urinary alkali excretion. In a multivariable model, urine pH and urine citrate were most discriminatory between CaP SF and CaOx SF (Receiver Operating Characteristic Area Under the Curve of 0.73 and 0.65 respectively). An increase in urine pH by 0.35, a decrease in urine citrate by 220 mg/day, a doubling of urine calcium, and female sex all independently doubled the risk of CaP compared to CaOx. High urine pH and hypocitraturia are two clinical parameters that distinguish the urine phenotype of CaP SF from CaOx SF. The alkalinuria is due to intrinsic differences in the kidney independent of intestinal alkali absorption and is accentuated in the female sex.

P53 deacetylation alleviates calcium oxalate deposition-induced renal fibrosis by inhibiting ferroptosis

Calcium oxalate (CaOx) stones are among the most common types of kidney stones and are associated with renal tubular damage, interstitial fibrosis, and chronic kidney disease. The mechanism of CaOx crystal-induced renal fibrosis remains unknown. Ferroptosis, a type of regulated cell death, is characterised by iron-dependent lipid peroxidation, and the tumour suppressor p53 is a key regulator of ferroptosis. In the present study, our results demonstrated that ferroptosis was significantly activated in patients with nephrolithiasis and hyperoxaluric mice as well as verified the protective effects of ferroptosis inhibition on CaOx crystal-induced renal fibrosis. Moreover, the single-cell sequencing database, RNA-sequencing, and western blot analysis revealed that the expression of p53 was increased in patients with chronic kidney disease and the oxalate-stimulated human renal tubular epithelial cell line, HK-2. Additionally, the acetylation of p53 was enhanced by oxalate stimulation in HK-2 cells. Mechanistically, we found that the induction of p53 deacetylation, owing to either the SRT1720-induced activation of deacetylase sirtuin 1 or the triple mutation of p53, inhibited ferroptosis and alleviated renal fibrosis caused by CaOx crystals. We conclude that ferroptosis is one of the critical mechanisms contributing to CaOx crystal-induced renal fibrosis, and the pharmacological induction of ferroptosis via sirtuin 1-mediated p53 deacetylation may be a potential target for preventing renal fibrosis in patients with nephrolithiasis.

Is hyperuricosuria really a risk factor for calcium oxalate stone formation?

In the seventies of the last century the term hyperuricosuric calcium oxalate (CaOx) urolithiasis has been coined. Allopurinol significantly decreased the recurrence in CaOx stone formers. Nevertheless, the role of hyperuricosuria has been questioned. Other mechanisms could have decreased the risk for recurrence. The underlying mechanisms are not clear. All these studies have been performed with only a limited number of patients. We studied n=2,480 consecutive patients with CaOx stones. For stone analysis, x-ray diffraction was used. The following parameters were examined in all patients: age, sex, body mass index (BMI), arterial blood pressure, stone episodes, diabetes mellitus; blood: creatinine, glucose, uric acid, calcium, sodium and potassium; urine: pH, volume, calcium, uric acid, citrate, ammonia and urea. According to the uric acid excretion (UAE), patients were divided into two groups: 1. UAE &gt; 4 mmol/d (hyperuricosuria), 2. UAE ≤ 4 mmol/d (normouricosuria). We observed significant correlations between UAE and sex, age, BMI, blood pressure, serum values of creatinine, sodium, urine volume, urine levels of calcium, citrate, urea and ammonia as well as serum calcium. No significant correlations were seen to the stone frequency (number of stone episodes), serum potassium and urine pH. In group 1 (n=741), there were 87.2% male and 12.8% female patients; in group 2 (n=1,739), the ratio was 60.6:39.4%. In the hyperuricosuric group, there were significantly more males than in the normouricosuric collective. Hyperuricosuric subjects had a higher BMI, higher blood levels of uric acid and glucose, a higher diuresis, a higher excretion of calcium, urea and ammonium, but a lower excretion of citrate than normouricosuric patients. All the other parameters did not show any significant differences. Especially the number of stone episodes was not different. We concluded that hyperuricosuria went along with many other risk factors for CaOx stone formation, but was not associated with an increased risk for recurrence. Therefore, UAE is not a prognostic marker in CaOx urolithiasis. It is unlikely that hyperuricosuria is a real risk factor for CaOx stone formation and hyperuricosuric CaOx urolithiasis is an own entity.