Recent advances on the mechanisms of kidney stone formation (Review).

Kidney stone disease is one of the oldest diseases known to medicine; however, the mechanisms of stone formation and development remain largely unclear. Over the past decades, a variety of theories and strategies have been developed and utilized in the surgical management of kidney stones, as a result of recent technological advances. Observations from the authors and other research groups suggest that there are five entirely different main mechanisms for kidney stone formation. Urinary supersaturation and crystallization are the driving force for intrarenal crystal precipitation. Randall's plaques are recognized as the origin of calcium oxalate stone formation. Sex hormones may be key players in the development of nephrolithiasis and may thus be potential targets for new drugs to suppress kidney stone formation. The microbiome, including urease­producing bacteria, nanobacteria and intestinal microbiota, is likely to have a profound effect on urological health, both positive and negative, owing to its metabolic output and other contributions. Lastly, the immune response, and particularly macrophage differentiation, play crucial roles in renal calcium oxalate crystal formation. In the present study, the current knowledge for each of these five aspects of kidney stone formation is reviewed. This knowledge may be used to explore novel research opportunities and improve the understanding of the initiation and development of kidney stones for urologists, nephrologists and primary care.

Effect of magnesium ammonium phosphate on the expression of adhesion molecules in sheep renal tubular epithelial cells.

Adhesion molecules play an important role in urinary calculus formation. The expressions of adhesion molecules in renal tubular has been reported in some animals. However, the role of adhesion molecules in the process of sheep urinary calculus formation is still unclear. The magnesium ammonium phosphate (MAP) is the main component of sheep urinary calculus. In this paper, the sheep renal tubular epithelial cells (RTECs) were isolated and treated with MAP, the expressions of osteopontin (OPN), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and apoptosis-related indicators caspase-3, Bcl-2 and Bax in RTECs were observed, the viability of RTECs was detected by Cell Counting Kit-8 (CCK-8). The levels of superoxide dismutase (SOD) and malondialdehyde (MDA), and the expressions of inflammatory factors Interleukin-6 (IL-6), Interleukin-1 (IL-1), Interleukin-17 (IL-17) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent (ELISA). The histopathological observation of kidney in urolithiasis sheep was made. The results showed that MAP could reduce the viability and SOD activity, enhance the activity of MDA significantly and promote the expressions of IL-1, IL-6, IL-17 and TNF-α of RTECs. By western blot and qPCR methods, the expressions of ICAM-1, VCAM-1 and OPN increased in 48 h. In addition, the expression of caspase-3 increased significantly and the ratio of Bcl-2/Bax reduced with exposure to MAP. The renal tissue structure was seriously damaged, the RTECs in urolithiasis sheep were degenerative and necrotic.

Biotransformation of struvite by Aspergillus niger: phosphate release and magnesium biomineralization as glushinskite.

Struvite (magnesium ammonium phosphate-MgNH4 PO4 ·6H2 O), which can extensively crystallize in wastewater treatments, is a potential source of N and P as fertilizer, as well as a means of P conservation. However, little is known of microbial interactions with struvite which would result in element release. In this work, the geoactive fungus Aspergillus niger was investigated for struvite transformation on solid and in liquid media. Aspergillus niger was capable of solubilizing natural (fragments and powder) and synthetic struvite when incorporated into solid medium, with accompanying acidification of the media, and extensive precipitation of magnesium oxalate dihydrate (glushinskite, Mg(C2 O4 ).2H2 O) occurring under growing colonies. In liquid media, A. niger was able to solubilize natural and synthetic struvite releasing mobile phosphate (PO4 3- ) and magnesium (Mg2+ ), the latter reacting with excreted oxalate resulting in precipitation of magnesium oxalate dihydrate which also accumulated within the mycelial pellets. Struvite was also found to influence the morphology of A. niger mycelial pellets. These findings contribute further understanding of struvite solubilization, element release and secondary oxalate formation, relevant to the biogeochemical cycling of phosphate minerals, and further directions utilizing these mechanisms in environmental biotechnologies such as element biorecovery and biofertilizer applications.

Understanding the biochemical characteristics of struvite bio-mineralising microorganisms and their future in nutrient recovery.

The biochemical properties of selected microorganisms (Bacillus pumilus, Brevibacterium antiquum, Myxococcus xanthus, Halobacterium salinarum and Idiomarina loihiensis), known for their ability to produce struvite through biomineralisation, were investigated. All five microorganisms grew at mesophilic temperature ranges (22-34 °C), produced urease (except I. loihiensis) and used bovine serum albumin as a carbon source. I. loihiensis was characterised as a facultative anaerobe able to use O2 and NO3 as an electron acceptor. A growth rate of 0.15 1/h was estimated for I. loihiensis at pH 8.0 and NaCl 3.5% w/v. The growth rates for the other microorganisms tested were 0.14-0.43 1/h at pH 7-7.3 and NaCl ≤1% w/v. All the microorganisms produced struvite, as identified by morphological and X-ray Powder Diffraction (XRD) analysis, under aerobic conditions. The biological struvite yield was between 1.5 and 1.7 g/L of media, the ortho-phosphate removal and recovery were 55-76% and 46-54%, respectively, the Mg2+ removal and recovery was 92-98% and 83-95%, respectively. Large crystals (>300 µm) were observed, with coffin-lid and long-bar shapes being the dominant morphology of biological struvite crystals. The characterisation of the biochemical properties of the studied microorganisms is critical for reactor and process design, as well as operational conditions, to promote phosphorus recovery from waste streams.

Development of a closed-loop process for fusel alcohol production and nutrient recycling from microalgae biomass.

Improving the economic feasibility is necessary for algae-based processes to achieve commercial scales for biofuels and bioproducts production. A closed-loop system for fusel alcohol production from microalgae biomass with integrated nutrient recycling was developed, which enables the reuse of nitrogen and phosphorus for downstream application and thus reduces the operational requirement for external major nutrients. Mixed fusel alcohols, primarily isobutanol and isopentanol were produced from Microchloropsis salina hydrolysates by an engineered E. coli co-culture. During the process, cellular nitrogen from microalgae biomass was converted into ammonium, whereas cellular phosphorus was liberated by an osmotic shock treatment. The formation of struvite from the liberated ammonium and phosphate, and the subsequent utilization of struvite to support M. salina cultivation was demonstrated. The closed loop system established here should help overcome one of the identified economic barriers to scale-up of microalgae production, and enhance the sustainability of microalgae-based chemical commodities production.

Phosphorus release and recovery from Fe-enhanced primary sedimentation sludge via alkaline fermentation.

Phosphorus release and recovery from Fe-based chemically enhanced primary sedimentation (CEPS) sludge via alkaline fermentation was investigated. The coagulation results showed that 78% of organic matter and 95% of phosphorus were concentrated from sewage into sludge with the optimum dosages of 25 mg/L FeCl3. The batch fermentation results revealed that 69.35% of the phosphorus in the Fe-sludge can be released and the maximum phosphorus concentration was 20.57 mg/L at pH 11. In the recovery stage, 90% of the P released in the fermented sludge supernatant was precipitated at a 2:1 ratio of magnesium to phosphorus and pH 11. The result of X-ray diffraction indicated that magnesium ammonium phosphate (MAP) was the major component of the precipitated solids. Thus, the present study provides an alternative option for phosphorus release and recovery as MAP from CEPS sludge via alkaline fermentation.

Factors associated with hematuric struvite crystalluria in cats.

OBJECTIVES: The aim of the study was to identify any dietary, signalment, geographic and clinical factors associated with hematuric struvite crystalluria (HSC) in a population of cats that visit general care veterinary hospitals in the USA. METHODS: In total, 4032 cats that had a first-time diagnosis of HSC and 8064 control cats with no history of hematuria or crystalluria were identified from medical records of all cats examined between 2007 and 2011 at 790 US veterinary hospitals. Extracted variables included age, sex, neuter status, breed, diet, urinalysis results and history of cystitis. Potential associations between these variables and HSC were estimated. RESULTS: Controlling for other factors, young cats fed a dry diet had an increased likelihood of HSC relative to young cats fed a non-dry diet. However, as age increased, the likelihood of HSC declined for cats fed a dry diet and increased for cats fed a non-dry diet. Moreover, the odds of HSC were significantly greater when cats were unneutered (vs neutered; odds ratio [OR] 45.52) or had a thin (vs heavy) body condition (OR 23.81), diagnosis of cystitis (OR 2.84), urine protein concentration >30 mg/dl (OR 4.72), alkaline (vs neutral) urine pH (OR 3.34), pyuria (OR 23.67) or bacteriuria (OR 2.24). CONCLUSIONS AND RELEVANCE: The present study provides estimates of the strengths of association between HSC and certain signalment and clinical characteristics of cats. This information could help clinicians to perform a more directed screening for struvite crystalluria in certain cat populations. Follow-up studies that build on the findings of this study could explore the clinical importance of HSC in cats.

Integrated process for anaerobically digested swine manure treatment.

An integrated three-step process was proposed for the treatment of the anaerobically digested swine manure (ADSM). The flocculation and struvite precipitation were used as the pre-treatment to remove the particles and reduce phosphorus to balance the condition for the algae growth. In the biological step, the 40% group (2.5× dilution) represented the optimal cultivation condition for the A + B co-cultivation, with the highest biomass concentration of 2.325 ±â€¯0.16 g/L and performed well with nutrients removal (COD: 9770 ±â€¯184 mg/L; TN: 235 ±â€¯5.4 mg/L; TP: 25.3 ±â€¯0.8 mg/L). 94.8% of the biomass from the 40% group could naturally settle down in 30 min which is good for harvest. The activated carbon adsorption was applied as the advanced treatment to resolve the issues with the dark color and residual compounds. After these processes, the removal efficiencies of COD, TN, TP and NH4-N reached 97.2%, 94.0%, 99.7% and 99.9%, respectively.

The role of pH on the biological struvite production in digested sludge dewatering liquors.

Struvite production mediated by bacteria has opened up a new route for phosphorus recovery from wastewater streams but its application to digested sludge dewatering liquors is not yet well understood. This study investigates the growth and biological struvite production of selected bacteria in wastewater liquors with pHs between 5.7 to 9.1. The bacterial growth was assessed through flow cytometry. Bacillus pumilus, Halobacterium salinarum and Brevibacterium antiquum remained viable at pHs between 5.7 to 9.1 but B. antiquum was able to grow at pHs between 7.3 to 7.8. Further analysis allowed the identification of crystals as struvite in tests between pH 7.3 to 8.3. All strains were capable of producing struvite at a range of pHs, but the highest production of 135-198 mg/L was observed for pHs between 7.3 to 8.3. At pHs > 8.3, precipitation of struvite and calcium compounds was observed in inoculated and non-inoculated tests. This study demonstrates that biological struvite production can occur at a wide range of pHs, hence significantly different from chemical struvite precipitation that occurs at pH > 8.3, making it a potentially viable process for phosphorus recovery as struvite from wastewater streams and sludge liquors without strict pH control.

Understanding the growth of the bio-struvite production Brevibacterium antiquum in sludge liquors.

Biological struvite (bio-struvite) production through biomineralization has been suggested as an alternative to chemically derived struvite production to recover phosphorus from wastewater streams. In this study, statistical experimental design techniques were used to find the optimal growth rate (µ) of Brevibacterium antiquum in sludge liquors. Acetate, oleic acid, NaCl, NH4-N, and Ca2+ were shown to affect the growth rate of B. antiquum. The growth rate reached 3.44 1/d when the bacteria were supplemented with 3.0% w/v NaCl and 1124 mg chemical oxygen demand/L as acetate. However, NaCl was found to hinder the biomineralization of bio-struvite. A two-stage experiment demonstrated that bio-struvite was produced in the presence of acetate. Bio-struvite production was confirmed with X-ray spectroscopy and crystal morphology (prismatic, tabular, and twinned crystal habit) through electron microscope analysis. The bio-struvite production was estimated by measuring phosphate content of the recovered precipitates, reaching 9.6 mg P/L as bio-struvite. Overall, these results demonstrated the optimal conditions required to achieve high growth rates as well as bio-struvite production with B. antiquum. The results obtained in this study could be used to develop a process to grow B. antiquum in wastewater streams in mixed cultures and recover phosphorus-rich products such as struvite.

Trace elements in struvite equine enteroliths: Concentration, speciation and influence of diet.

Equine enteroliths ∼1.5cm in diameter were collected from an Arabian horse in Louisville, Kentucky, United States. Scanning electron microscopy (SEM) and light microscope imaging of a sectioned enterolith showed two distinct regions of concentric growth outward from the central nidus, a small pebble. After initial growth, acidic colonic fluids permeated the stone inducing recrystallization and alteration of crystals closest to the nidus. A second growth event, when mineral crystallization was again favorable, produced an outer region of unaltered crystals at the rim. The mineral was identified as struvite (MgNH4PO4∙6H2O) by powder X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy. Elemental analysis confirmed concentrations of P, Mg and N consistent with the struvite composition, and detected trace elements Fe (1050-1860mgkg-1), Mn (262-280mgkg-1) and Zn (197-238mgkg-1). All elements were traced to dietary sources, with the Fe:Mn:Zn ratio of the enterolith consistent with that of the horse feed. X-ray absorption fine structure (XAFS) spectroscopy at the Zn K-edge revealed distorted ZnO4 tetrahedra located between crystallographic planes in the struvite structure forming bidentate linkages to struvite phosphate groups. Emplacement of Zn in structural cavities likely occurs during struvite crystallization. Trace elements and organic impurities increase susceptibility of the enterolith to heat-induced decomposition relative to pure struvite, which could be a consideration for treatment. Results reveal enterolith growth processes, composition and mechanisms of trace metal accumulation that can inform management and prevention of equine enteroliths.

Agronomic Effectiveness of Granulated and Powdered P-Exchanged Mg-Al LDH Relative to Struvite and MAP.

Layered double hydroxides (LDHs) used to recover P from wastewater have recently been proposed as new slow-release fertilizers. Here, the use of P-exchanged Mg-Al LDHs as powdered or granulated fertilizer is explored and compared with monoammonium phosphate (MAP), a fully water-soluble fertilizer, and with struvite, a recycled phosphate fertilizer with lower solubility. First, these three fertilizers were compared in a 100-day incubation experiment using P diffusion visualization and chemical analysis to assess P release from either granules or powdered fertilizer in three different soils. By the end of the incubation, 74-90% of P remained within the LDH granule, confirming a slow release. Second, a pot experiment was performed with wheat (Triticum aestivum) in an acid and a calcareous soil. The granular treatment resulted in a considerably higher P uptake for MAP compared to LDH and struvite. For the powder treatments, the P uptake was less than for granular MAP and was largely unaffected by the chemical form. The LDHs and struvite showed a lower agronomic effectiveness than granular MAP, but the benefits of their use in P recycling, potential residual value, and environmental benefits may render these slow-release fertilizers attractive.

Bacterially Induced Formation of Infectious Urinary Stones: Recent Developments and Future Challenges.

Infectious urinary stones make a serious medical problem concerning up to 20% of population. Additionally, recurrence after treatment reaches 50%. The formation of infectious urinary stones is connected with urinary tract infection with various bacteria. These are mainly the bacteria from Proteus species which have been isolated in 70% of bacteriainduced urinary stone cases. These microorganisms produce urease - a bacterial enzyme which plays a principal role in the crystallization process and is one of the main virulence parameters of these bacteria. The most common solid components of infectious urinary stones are the crystals of struvite and amorphous carbonate apatite. The formation of this kind of stones involves two main processes. The first one is the nucleation process of solid phases and the second is the aggregation of the precipitated phases, both crystalline and amorphous. In recent years, both these processes have been deeply investigated. In particular, different active compounds have been reported as potentially novel therapeutic agents to prevent or inhibit the formation of infectious urinary stones. In addition, there is rich scientific evidence regarding the chemical mechanisms of inhibitory effect of these active compounds. In recent years, specific interior and exterior structure of struvite and its porous nature have also been reported. In this article, we summarize and discuss recent development in this field of research. The paper concludes with future goals and challenges.

Whole-cell Proteus mirabilis urease inhibition by aminophosphinates for the control of struvite formation.

The study evaluated the in vitro impact of a series of aminophosphinic urease inhibitors on Proteusmirabilis. The group of compounds comprised structurally diverse analogues of diamidophosphate built on an N-C-P scaffold. The influence of urease inhibition on urea-splitting activity was assessed by whole-cell pH-static kinetic measurements. The potential to prevent struvite formation was determined by monitoring changes in pH and ionic composition of artificial urine medium during P. mirabilis growth. The most active compounds exhibited stronger positive effect on urine stability than the acknowledged inhibitor acetohydroxamic acid. The high anti-ureolytic and pH-stabilizing effect of urease inhibitors 4 and 14 was well correlated with their reported kinetic properties against pure urease from P. mirabilis (Ki values of 0.62±0.09 and 0.202±0.057 µM, respectively, compared to 5.7±0.4 µM for acetohydroxamic acid). The effect of repressed ureolysis upon the viability of Proteus cells was studied using MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] metabolic efficiency assay and LIVE/DEAD fluorescent staining. Most of the compounds caused whole-cell dehydrogenase activity loss; four structures (1, 2, 4 and 14) reduced the culture viability by nearly 70 % at 1 mM concentration. Results of dual fluorescent staining suggested that besides urea-splitting prevention, the structures additionally exerted an outer-membrane-destabilizing effect.

Influence of lime on struvite formation and nitrogen conservation during food waste composting.

This study aimed at investigating the feasibility of supplementing lime with struvite salts to reduce ammonia emission and salinity consequently to accelerate the compost maturity. Composting was performed in 20-L bench-scale reactors for 35days using artificial food waste mixed with sawdust at 1.2:1 (w/w dry basis), and Mg and P salts (MgO and K2HPO4, respectively). Nitrogen loss was significantly reduced from 44.3% to 27.4% during composting through struvite formation even with the addition of lime. Lime addition significantly reduced the salinity to less than 4mS/cm with a positive effect on improving compost maturity. Thus addition of both lime and struvite salts synergistically provide advantages to buffer the pH, reduce ammonia emission and salinity, and accelerate food waste composting.

Combined use of nitrification inhibitor and struvite crystallization to reduce the NH3 and N2O emissions during composting.

Struvite crystallization (SCP) is combined with a nitrification inhibitor (dicyandiamide, DCD) to mitigate the NH3 and N2O emission during composting. The MgO and H3PO4 were added at a rate of 15% (mole/mole) of initial nitrogen, and the DCD was added at rates of 0%, 2.5%, 5.0%, 7.5% and 10% (w/w) of initial nitrogen respectively. Results showed that the combination use of SCP and DCD was phytotoxin free. The SCP could significantly reduce NH3 losses by 45-53%, but not the DCD. The DCD significantly inhibits nitrification when the content was higher than 50mgkg(-1), and that could reduce the N2O emission by 76.1-77.6%. The DCD degraded fast during the thermophilic phase, as the nitrification will be inhibited by the high temperature and high free ammonia content in this stage, the DCD was suggested to be applied in the maturing periods by 2.5% of initial nitrogen.

Growth of mono- and mixed cultures of Nannochloropsis salina and Phaeodactylum tricornutum on struvite as a nutrient source.

The suitability of crude and purified struvite (MgNH4PO4), a major precipitate in wastewater streams, was investigated for renewable replacement of conventional nitrogen and phosphate resources for cultivation of microalgae. Bovine effluent wastewater stone, the source of crude struvite, was characterized for soluble N/P, trace metals, and biochemical components and compared to the purified mineral. Cultivation trials using struvite as a major nutrient source were conducted using two microalgae production strains, Nannochloropsis salina and Phaeodactylum tricornutum, in both lab and outdoor pilot-scale raceways in a variety of seasonal conditions. Both crude and purified struvite-based media were found to result in biomass productivities at least as high as established media formulations (maximum outdoor co-culture yield ∼20±4gAFDW/m(2)/day). Analysis of nutrient uptake by the alga suggest that struvite provides increased nutrient utilization efficiency, and that crude struvite satisfies the trace metals requirement and results in increased pigment productivity for both microalgae strains.