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Pretreatment with BS168 prevents increased calcium oxalate crystal aggregation and adhesion to renal epithelial cells. (A) Crystal aggregates were visualized by light microscopy after MDCK monolayers were pretreated with MEM with or without BS168 or UTI89 (positive control), followed by artificial urine with or without 5 mg/ml CaOx monohydrate. Bars, 200 m. (B) The amounts of CaOx monohydrate crystals adhering to MDCK cells did not differ significantly between the CaOx groups with or without BS168 pretreatment. (C) Average crystal aggregate sizes did not differ significantly between the CaOx groups with or without BS168 pretreatment. Each point represents the average from three fields of view (FOV) in a single well of a 6-well plate, and the median for each group is indicated by a horizontal line. Three technical and three biological replicates were performed (n 9). Significance was determined by a Kruskal-Wallis test with Dunn's multiple comparisons (*, P 0.05; ***, P 0.001; ns, not significant).
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Kidney stone disease is a morbid condition that is increasing in prevalence, with few nonsurgical treatment options. The majority of stones are composed of calcium oxalate. Unlike humans, some microbes can break down oxalate, suggesting that microbial therapeutics may provide a novel treatment for kidney stone patients. This study demonstrated that...
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... with BS168 prevents increased calcium oxalate crystal aggregation and adhesion to renal epithelial cells. MDCK renal epithelial cells were utilized to assess the effect of BS168 pretreatment on the adhesion and aggregation of CaOx monohydrate crystals in artificial urine (Fig. 5A). Cells that were pretreated with BS168 prior to CaOx exhibited no more crystal adhesion than vehicle control cells, in contrast to cells that were pretreated with uropathogenic Escherichia coli (UPEC) strain UTI89, which was utilized as a positive control (Fig. 5B). Average crystal sizes did not differ significantly between the CaOx ...
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... on the adhesion and aggregation of CaOx monohydrate crystals in artificial urine (Fig. 5A). Cells that were pretreated with BS168 prior to CaOx exhibited no more crystal adhesion than vehicle control cells, in contrast to cells that were pretreated with uropathogenic Escherichia coli (UPEC) strain UTI89, which was utilized as a positive control (Fig. 5B). Average crystal sizes did not differ significantly between the CaOx groups with or without BS168 pretreatment, indicating that BS168 did not encourage aggregation (Fig. ...
Context 3
... than vehicle control cells, in contrast to cells that were pretreated with uropathogenic Escherichia coli (UPEC) strain UTI89, which was utilized as a positive control (Fig. 5B). Average crystal sizes did not differ significantly between the CaOx groups with or without BS168 pretreatment, indicating that BS168 did not encourage aggregation (Fig. ...
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Aims
This study estimated the prevalence of low eGFR in those without known hypertension, T2DM or heavy proteinuria as a surrogate marker for chronic kidney disease of unknown cause (CKDu) among adults in the North of Peru.
Methods
A cross-sectional study was conducted following the Disadvantaged Populations eGFR Epidemiology (DEGREE) Study protoc...
Citations
... Calcium oxalate is the major component of kidney stones, therefore ideal probiotic bacteria are efficient in oxalate degradation, such as Oxalobacter formigenes 225 and B. subtilis. B. subtilis strain 168 has been presented as a novel probiotic therapy as it has been shown to break down the oxalate in kidney stones in a Drosophila model 226 . The oxalate decarboxylase (OxDC) enzyme derived from B. subtilis can also be used to engineer other bacteria as probiotics for the treatment of kidney stones. ...
With the construction of the International Space Station, humans have been continuously living and working in space for 22 years. Microbial studies in space and other extreme environments on Earth have shown the ability for bacteria and fungi to adapt and change compared to "normal" conditions. Some of these changes, like biofilm formation, can impact astronaut health and spacecraft integrity in a negative way, while others, such as a propensity for plastic degradation, can promote self-sufficiency and sustainability in space. With the next era of space exploration upon us, which will see crewed missions to the Moon and Mars in the next 10 years, incorporating microbiology research into planning, decision-making, and mission design will be paramount to ensuring success of these long-duration missions. These can include astronaut microbiome studies to protect against infections, immune system dysfunction and bone deterioration, or biological in situ resource utilization (bISRU) studies that incorporate microbes to act as radiation shields, create electricity and establish robust plant habitats for fresh food and recycling of waste. In this review, information will be presented on the beneficial use of microbes in bioregenerative life support systems, their applicability to bISRU, and their capability to be genetically engineered for biotechnological space applications. In addition, we discuss the negative effect microbes and microbial communities may have on long-duration space travel and provide mitigation strategies to reduce their impact. Utilizing the benefits of microbes, while understanding their limitations, will help us explore deeper into space and develop sustainable human habitats on the Moon, Mars and beyond.
... In contrast, Agxt is also downregulated and hypermethylated in hepatocytes from patients with nonalcoholic fatty liver disease (NAFLD), providing a mechanistic explanation for the increased risk of kidney stones and CKD in NAFLD patients (51). Another study indicated that Bacillus subtilis 168 (BS168) degrades oxalate and reduces the severity of calculi, also providing a new microbial therapy for stone treatment (52). Moreover, Li et al. indicated that Obcordata A (OA), a polyoxygestrel glycoside, prevents kidney stones through the mitogen-activated protein kinase (MAPK) pathway. ...
... This provides a new theoretical basis for the prevention and treatment of kidney stones (82). It is known that phospholipase A2(PLA2) can induce mitochondrial damage to produce excess ROS, while BS168 can blunt PLA2 to regulate ROS metabolism and thus affect stone formation (52). Increased calcium and/or oxalate in renal tubular solution may activate nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in renal epithelial cells via the renin-angiotensin system and trigger the production of ROS(14978165 (83)). ...
Kidney stone disease (KSD) is one of the earliest medical diseases known, but the mechanism of its formation and metabolic changes remain unclear. The formation of kidney stones is a extensive and complicated process, which is regulated by metabolic changes in various substances. In this manuscript, we summarized the progress of research on metabolic changes in kidney stone disease and discuss the valuable role of some new potential targets. We reviewed the influence of metabolism of some common substances on stone formation, such as the regulation of oxalate, the release of reactive oxygen species (ROS), macrophage polarization, the levels of hormones, and the alternation of other substances. New insights into changes in substance metabolism changes in kidney stone disease, as well as emerging research techniques, will provide new directions in the treatment of stones. Reviewing the great progress that has been made in this field will help to improve the understanding by urologists, nephrologists, and health care providers of the metabolic changes in kidney stone disease, and contribute to explore new metabolic targets for clinical therapy.
... After 24 h, these flies were transferred to either normal or lithogenic medium containing 0.1% (w/v) sodium oxalate. This alternating process was repeated until day 10, when D. melanogaster Malpighian tubules were dissected [26]. Briefly, flies were anesthetized using CO 2 and dissected in cold phosphate buffer solution (PBS) using a Nikon SMZ800N stereomicroscope. ...
Kidney stone disease affects nearly one in ten individuals and places a significant economic strain on global healthcare systems. Despite the high frequency of stones within the population, effective preventative strategies are lacking and disease prevalence continues to rise. Osteopontin (OPN) is a urinary protein that can inhibit the formation of renal calculi in vitro. However, the efficacy of OPN in vivo has yet to be determined. Using an established Drosophila melanogaster model of calcium oxalate urolithiasis, we demonstrated that a 16-residue synthetic OPN phosphopeptide effectively reduced stone burden in vivo. Oral supplementation with this peptide altered crystal morphology of calcium oxalate monohydrate (COM) in a similar manner to previous in vitro studies, and the presence of the OPN phosphopeptide during COM formation and adhesion significantly reduced crystal attachment to mammalian kidney cells. Altogether, this study is the first to show that an OPN phosphopeptide can directly mitigate calcium oxalate urolithiasis formation in vivo by modulating crystal morphology. These findings suggest that OPN supplementation is a promising therapeutic approach and may be clinically useful in the management of urolithiasis in humans.
... Following 2 courses of bacterial supplementation and toxin exposure (Fig. 4a), oxidative stress was assessed in the Drosophila. Confocal microscopy was used to visualize ROS in the Malpighian tubules that were dissected (75) and stained (76) with 29,79-dichlorofluorescein (Sigma) as previously described. Images were acquired using a Nikon A1R confocal microscope and the associated NIS-Elements software. ...
Renal impairment associated with chronic kidney disease (CKD) causes the buildup of uremic toxins that are deleterious to patient health. Current therapies that manage toxin accumulation in CKD offer an incomplete therapeutic effect against toxins such as para-cresol (p-cresol) and p-cresyl sulfate. Probiotic therapies can exploit the wealth of microbial diversity to reduce toxin accumulation. Using in vitro culture techniques, strains of lactobacilli and bifidobacteria from a 24-strain synbiotic were investigated for their ability to remove p-cresol. Four strains of bifidobacteria internalized p-cresol from the extracellular environment. The oral supplementation of these toxin-clearing probiotics was more protective than control strains in a Drosophila melanogaster toxicity model. Bifidobacterial supplementation was also associated with higher abundance of lactobacilli in the gut microbiota of p-cresol-exposed flies. The present findings suggest that these strains might reduce p-cresol in the gut in addition to increasing the prevalence of other beneficial bacteria, such as lactobacilli, and should be tested clinically to normalize the dysbiotic gut microbiota observed in CKD patients.
IMPORTANCE Chronic kidney disease (CKD) affects approximately 10% of the global population and has limited treatment options. The accumulation of gut microbiota-derived uremic toxins, such as para-cresol (p-cresol) and p-cresyl sulfate, is associated with the onset of comorbidities (i.e., atherosclerosis and cognitive disorders) in CKD. Unfortunately, dialysis, the gold standard therapy is unable to remove these toxins from the bloodstream due to their highly protein-bound nature. Some strains of Bifidobacterium have metabolic properties that may be useful in managing uremic toxicity. Using a Drosophila model, the present work highlights why dosing with certain probiotic strains may be clinically useful in CKD management.
... For example, natto, a Japanese soy product fermented by Bacillus species is a potent source of vitamin K2, with proven benefits toward bone health [104,105]. These Bacillus sp. may also hold promise as potential probiotics in mitigating calcium oxalate kidney stones [106], which are a significant threat to long-term space travel [65]. In another study by Hati et al. [107], Lactobacillus isolates from traditional fermented Indian foods demonstrated potent B vitamin and shortchain fatty acid production, both of which can become dysregulated in astronauts [108] and the simulated space environment [109]. ...
The microbiota is important for immune modulation, nutrient acquisition, vitamin production, and other aspects for long-term human health. Isolated model organisms can lose microbial diversity over time and humans are likely the same. Decreasing microbial diversity and the subsequent loss of function may accelerate disease progression on Earth, and to an even greater degree in space. For this reason, maintaining a healthy microbiome during spaceflight has recently garnered consideration. Diet, lifestyle, and consumption of beneficial microbes can shape the microbiota, but the replenishment we attain from environmental exposure to microbes is important too. Probiotics, prebiotics, fermented foods, fecal microbiota transplantation (FMT), and other methods of microbiota modulation currently available may be of benefit for shorter trips, but may not be viable options to overcome the unique challenges faced in long-term space travel. Novel fermented food products with particular impact on gut health, immune modulation, and other space-targeted health outcomes are worthy of exploration. Further consideration of potential microbial replenishment to humans, including from environmental sources to maintain a healthy microbiome, may also be required.
... For the phytochemical oxalate, the gut bacterium Ishikawaella capsulata in stinkbug Megacopta punctatissima encodes genes for oxalate decarboxylase, suggesting the possible role of the bacterium in oxalate detoxification [53]. In human, calcium oxalate is formed if the food-derived oxalate cannot be metabolized, which can result in kidney stone disease, so the identification of insect bacteria able to degrade oxalate may act as a novel treatment for kidney stone patients [54] (Table 1). Encodes genes of oxalate decarboxylase (by whole-genome shotgun sequencing) [53] Apart from detoxification roles, some insect bacteria can convert phytochemicals into pheromone compounds and thus influence the chemical communication of host insects [56]. ...
The diversity and high adaptability of insects are heavily associated with their symbiotic microbes, which include bacteria, fungi, viruses, protozoa, and archaea. These microbes play important roles in many aspects of the biology and physiology of insects, such as helping the host insects with food digestion, nutrition absorption, strengthening immunity and confronting plant defenses. To maintain normal development and population reproduction, herbivorous insects have developed strategies to detoxify the substances to which they may be exposed in the living habitat, such as the detoxifying enzymes carboxylesterase, glutathione-S-transferases (GSTs), and cytochrome P450 monooxygenases (CYP450s). Additionally, insect symbiotic bacteria can act as an important factor to modulate the adaptability of insects to the exposed detrimental substances. This review summarizes the current research progress on the role of insect symbiotic bacteria in metabolizing phytochemicals and agrochemicals (insecticides and herbicides). Given the importance of insect microbiota, more functional symbiotic bacteria that modulate the adaptability of insects to the detrimental substances to which they are exposed should be identified, and the underlying mechanisms should also be further studied, facilitating the development of microbial-resource-based pest control approaches or protective methods for beneficial insects.
... However, they are not the only probiotics, that can degrade oxalate, Bacillus subtilis was also identified as an oxalate degrading bacterium in vitro since 2000 [53]. A recent in vivo study showed that Bacillus subtilis 168 (BS168) reduces CaOx stones in the MTs, increases the lifespan, and prevents the oxalate-induced microbiota dysbiosis of the fruit fly [54]. ...
Kidney stone disease is a global problem affecting about 12% of the world population. Novel treatments to control this disease have a huge demand. Here we argue that the fruit fly, as an emerging kidney stone model, can provide a platform for the discovery of new drugs. The renal system of fruit fly (Malpighian tubules) is similar to the mammalian renal tubules in both function and structure. Different fruit fly models for different types of kidney stones including calcium oxalate (CaOx) stones, xanthine stones, uric acid stone, and calcium phosphate (CaP) stones have been successfully established through dietary or genetic approaches in the last ten years, notably improved our understanding of the formation mechanisms of kidney stone diseases. The fruit fly CaOx stones model, which is mediated by treatment with dietary lithogenic agents, is also one of the most potential models for drug development. Various potential antilithogenic agents have been identified using this model, including new chemical compounds and medicinal plants. The fruit fly kidney stone models also afford opportunities to study the therapeutic mechanism of these drugs in deeper.
... The ability to prevent the formation of urinary stones is also demonstrated by B. subtills. Al and colleagues [114], using the Drosophila melanogaster model of urolithiasis as a high-throughput screening platform to evaluate the therapeutic potential of oxalatedegrading bacteria in calcium oxalate nephrolithiasis, found that Bacillus subtilis 168 (BS168) is promising candidate for the prevention of oxalate-induced microbiota dysbiosis. The studied BS168 was characterized by preferential growth in high oxalate concentrations and an ability to rapidly and stably colonize the D. melanogaster intestinal tract for as long as five days. ...
... These results were confirmed by in vitro study, which demonstrated that established MDCK renal cells treated with BS168 showed reduced adhesion and aggregation of calcium oxalate crystals. Thus, BS168 could represent a novel therapeutic adjunct to reduce the incidence of recurrent calcium oxalate nephrolithiasis in high-risk patients [114]. ...
Nephrolithiasis ranks third among urological diseases in terms of prevalence, making up about 15% of cases. The continued increase in the incidence of nephrolithiasis is most probably due to changes in eating habits (high protein, sodium, and sugar diets) and lifestyle (reduced physical activity) in all developed countries. Some 80% of all kidney stones cases are oxalate urolithiasis, which is also characterized by the highest risk of recurrence. Frequent relapses of nephrolithiasis contribute to severe complications and high treatment costs. Unfortunately, there is no known effective way to prevent urolithiasis at present. In cases of diet-related urolithiasis, dietary changes may prevent recurrence. However, in some patients, the condition is unrelated to diet; in such cases, there is evidence to support the use of stone-related medications. Interestingly, a growing body of evidence indicates the potential of the microbiome to reduce the risk of developing renal colic. Previous studies have primarily focused on the use of Oxalobacterformigenes in patients with urolithiasis. Unfortunately, this bacterium is not an ideal probiotic due to its antibiotic sensitivity and low pH. Therefore, subsequent studies sought to find bacteria which are capable of oxalate degradation, focusing on well-known probiotics including Lactobacillus and Bifidobacterium strains, Eubacterium lentum, Enterococcus faecalis, and Escherichia coli.
... This is in direct contradiction to a 2018 study by Tasian et al. [24], who reported that broad-spectrum antibiotics were associated with a diagnosis of kidney stones. Together, these data suggest that the effect of antibiotics on CaOx stone formation is not direct; instead, it might be derived from indirect interactions, whereby antibiotics induce inflammation in resident host cells or alter the microbiota [26,27]. Additionally, using our model we found that ciprofloxacin and TMP/SMX did not increase stone formation as previously described [7,8]. ...
Objective:
Kidney stones are a common medical condition that is increasing in prevalence worldwide. Approximately, ∼80% of urinary calculi are composed of calcium oxalate (CaOx). There is a growing interest toward identifying therapeutic compounds that can inhibit the formation of CaOx crystals. However, some chemicals (e.g., antibiotics and bacterial metabolites) may directly promote crystallization. Current knowledge is limited regarding crystal promoters and inhibitors. Thus, we have developed an in vitro gel-based diffusion model to screen for substances that directly influence CaOx crystal formation.
Materials and methods:
We used double diffusion of sodium oxalate and calcium chloride-loaded paper disks along an agar medium to facilitate the controlled formation of monohydrate and dihydrate CaOx crystals. A third disk was used for the perpendicular diffusion of a test substance to assess its influence on CaOx crystal formation.
Results:
We confirmed that citrates and magnesium are effective inhibitors of CaOx crystals. We also demonstrated that 2 strains of uropathogenic Escherichia coli are able to promote crystal formation. While the other tested uropathogens and most antibiotics did not change crystal formation, ampicillin was able to reduce crystallization.
Conclusion:
We have developed an inexpensive and high-throughput model to evaluate substances that influence CaOx crystallization.
Kidney stone disease (KSD) (alternatively nephrolithiasis or urolithiasis) is a global healthcare problem that affects almost all of developed and developing countries. Its prevalence has been continuously increasing with a high recurrence rate after stone removal. Although effective therapeutic modalities are available, preventive strategies for both new and recurrent stones are required to reduce physical and financial burdens of KSD. To prevent kidney stone formation, its etiology and risk factors should be firstly considered. Low urine output and dehydration are the common risks for all stone types, whereas hypercalciuria, hyperoxaluria and hypocitraturia are the major risks of calcium stone. In this article, up-to-date knowledge on strategies (nutrition-based mainly) to prevent KSD is provided. Important roles of fluid intake (2.5-3.0 L/day), diuresis (>2.0-2.5 L/day), lifestyle and habit modifications (e.g., maintain normal body mass index, fluid compensation for working in high-temperature environment, and avoid cigarette smoking), and dietary management (e.g., sufficient calcium at 1,000-1,200 mg/day, limit sodium at 2 g/day or 3-5 g/day of NaCl, limit oxalate-rich foods, avoid vitamin C and vitamin D supplements, limit animal proteins to 0.8-1.0 g/kg body weight/day but increase plant proteins in calcium and uric acid stone patients and those with hyperuricosuria, increase proportion of citrus fruits, and consider lime powder supplementation) are summarized. Moreover, uses of natural bioactive products (e.g., caffeine, epigallocatechin gallate, and diosmin), medications (e.g., thiazides, alkaline citrate, other alkalinizing agents, and allopurinol), bacterial eradication, and probiotics are also discussed.
