Oxalate Crystals Research Articles

Agrobacterium rhizogenes-mediated marker-free transformation and gene editing system revealed that AeCBL3 mediates the formation of calcium oxalate crystal in kiwifruit

The transformation and gene editing of the woody species kiwifruit are difficult and time-consuming. The fast and marker-free genetic modification system for kiwifruit has not been developed yet. Here, we establish a rapid and efficient marker-free transformation and gene editing system mediated by Agrobacterium rhizogenes for kiwifruit. Moreover, a removing-root-tip method was developed to significantly increase the regeneration efficiency of transgenic hairy roots. Through A. rhizogenes-mediated CRISPR/Cas9 gene editing, the editing efficiencies of CEN4 and AeCBL3 achieved 55 and 50%, respectively. And several homozygous knockout lines for both genes were obtained. Our method has been successfully applied in the transformation of two different species of kiwifruit (Actinidia chinensis ‘Hongyang’ and A.eriantha ‘White’). Next, we used the method to study the formation of calcium oxalate (CaOx) crystals in kiwifruit. To date, little is known about how CaOx crystal is formed in plants. Our results indicated that AeCBL3 overexpression enhanced CaOx crystal formation, but its knockout via CRISPR/Cas9 significantly impaired crystal formation in kiwifruit. Together, we developed a fast maker-free transformation and highly efficient CRISPR-Cas9 gene editing system for kiwifruit. Moreover, our work revealed a novel gene mediating CaOx crystal formation and provided a clue to elaborate the underlying mechanisms.Graphical abstract

Ethylene glycol toxicity in two calves

Burdur Mehmet Akif University Veterinary Faculty Animal Hospital was brought from livestock to 3 calves aged between 15 and 20 days with complaints of weakness, dehydration, inability to get up and nervous findings. Two of them failed the treatment and were brought to the pathology department after being euthanized. Both animals were macroscopically dehydrated, cachectic, and had hair loss in some parts of the body. There was 1-2 liters of white light-colored transparent fluid in the abdominal cavity. Many organs and serous membranes in the abdominal cavity were hyperemic. Histopathological examination revealed bleeding, edema and degenerative changes in the heart, lungs and kidneys. Calcium oxalate crystals were found in many tubuler lumens, especially the proximal convoluted tubules in the kidneys. In the light of clinical, macroscopic, and histopathological findings, the case was diagnosed with ethylene glycol toxicity. Ethylene glycol toxicity is often seen in cats and dogs living near industrial or auto car repair shops, and in animals that accidentally drink antifreeze waste. This case report is the first case of ethylene glycol toxicity reported in livestock calves in Türkiye.

3-Demethyl-2-geranyl-4-prenylbellidifoline, a natural xanthone with diuretic and kidney protective properties.

The diuretic and kidney protective effect of the 3-demethyl-2-geranyl-4-prenylbellidifoline (DGP) were evaluated in rats. The normotensive (NTR) and spontaneously hypertensive rats (SHR) received, once a day for 7 days, oral treatment with DGP (0.1 mg/kg), hydrochlorothiazide (10 mg/kg), or vehicle (10 ml/kg). Urine, blood, and kidney samples were collected for further analysis. The urine and Na+ elimination content were significantly higher in the groups that received DGP. Furthermore, a Ca2+-sparing action was detected in the urine of DGP-treated groups, which was consistent with the reduction in calcium oxalate crystal formation. Relevantly, the treatment did not change the parameters examined in the blood. Concerning the renal analyses, DGP treatment recovered the morphological damages of the kidney corpuscle area of SHR. In addition to the differences observed between the NTR and SHR vehicle groups, DGP augmented the amount of reduced glutathione and the activity of glutathione S-transferase GST while reducing the catalase and N-acetyl-β-D-glucosaminidase activity and nitrite levels. Together, this study displayed the prolonged diuretic action of DGP and its natriuretic, Ca2+-sparing, and antiurolytic effects. The antioxidative and anti-inflammatory effects of DGP were evidenced in SHR kidneys, opening perspectives for further studies regarding the benefits of this xanthone.

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.

Metabolomics analysis reveals a protective effect of hydroxycitric acid on calcium oxalate-induced kidney injury.

Prior research has indicated that hydroxycitric acid (HCA) can impede the formation of calcium oxalate (CaOx) crystals, yet the specific mechanisms underlying its therapeutic effects remain unclear. In this study, we delved into the protective effects of HCA against glyoxylate-induced renal stones in rats and sought to elucidate the underlying metabolic pathways. Forty rats were randomly assigned to five groups: control group, model group, L-HCA-treated group, M-HCA-treated group, and H-HCA-treated group. Von Kossa staining was conducted on renal sections, and blood urea nitrogen and serum creatinine were determined by biochemical analysis. Meanwhile, body weight and urine volume were also measured. We subjected urine samples from the rats to analysis using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Next, we employed a metabolomic approach to scrutinize the metabolic profiles of each group. HCA significantly reduced blood urea nitrogen and serum creatinine, and increased body weight and urine volume. It also reduced CaOx crystal deposition. A total of 24 metabolites, exhibiting a significant reversal pattern following HCA administration, were identified as urine biomarkers indicative of HCA's preventive effects against CaOx crystal-induced renal injury. These metabolites are primarily associated with glycine, serine, and threonine metabolism; phenylalanine metabolism; tricarboxylic acid cycle; taurine and hypotaurine metabolism; and tryptophan metabolism. It was demonstrated that HCA has a protective effect against CaOx crystal-induced kidney injury in rats by modulating various metabolic pathways. Additionally, results suggest that HCA holds promise as a potential clinical therapeutic drug for both the prevention and treatment of renal stones.

Nickel as a modifier of calcium oxalate: an in situ liquid cell TEM investigation of nucleation and growth.

Despite extensive research on the nucleation and growth of calcium oxalate (CaOx) crystals, there are still several challenges and unknowns that remain. In particular, the role of trace metal elements in the promotion or inhibition of CaOx crystals is not well understood. In the present study, in situ graphene liquid cell transmission electron microscopy (in situ GLC TEM) was used to observe real-time, nanoscale transformations of CaOx crystals in the presence of nickel ions (Ni2+). The results showed that Ni2+ form Ni-water complexes, acting as a shape-directing species, generating a unique morphology and altering growth kinetics. Transient adsorption of Ni-water complexes resulted in a metastable phase formation of calcium oxalate trihydrate. Atomistic molecular dynamics simulations confirmed that Ni2+ acts as a weak inhibitor which slows down the CaOx crystallization, elucidating that Ni2+ impacts small-sized CaOx clusters by bringing more water into the clusters. This work highlighted the intricacies behind the effect of Ni2+ on CaOx biomineralization that were made possible to discern using in situ GLC TEM.

COMPARATIVE ANALYSIS OF THE ANATOMICAL AND MORPHOLOGICAL FEATURES OF RHEUM TATARICUM L. PLANTS FROM DIFFERENT ECOPOPULATIONS

There are about 60 species of the genus Rheum L. (Polygonaceae) distributed mainly in temperate and subtropical Asia, with the largest number (38 species, including 19 endemics) recorded in China. All species of the genus are used as medicinal plants. According to modern pharmacology, the most important property of the rhubarb root is regulation of the stomach and intestine function, due to its laxa-tive effect; rhubarb also has antipyretic properties and antioxidant activity. The high content of dietary fibers provides protection against cardiovascular diseases, and vitamin K is valuable for the prevention of osteoporosis. However, the active ingredients and their mechanism of action have not been sufficiently studied.Rheum tataricum L., known for its high resistance to drought, salinity, and lack of nutrients is the least studied species of wild rhubarb. The whole plant, and especially the leaves, contains several toxic compounds, in particular anthraquinone glycosides and calcium oxalate crystals. Here we present the results of a comparative analysis of the anatomical and morphological structure of Rh. tataricum plants from ecopopulations growing in the Balkhash and Karatal districts of the Almaty region. We identified the differences due to the weather and climatic conditions of the ecopopulations. It is important to note that many idioblasts containing calcium oxalate were found in the rhizomes of Rh. tataricum, which prevent animals from eating plants and getting poisoned. The number of idioblasts in the central circle of the Rh. tataricum rhizome was smaller in the Balkhash district than in the Karatal district, while the idioblasts were larger in size in the former district. The medicinal and nutritional value of Rh. tataricum growing in a semi-desert environment indicates a high potential of the species in plant breeding, medicine, and as a food source.

Alocasia odora poisoning due to calcium oxalate needle crystals in Japan.

Objective: To elucidate the actual circumstances of damage caused by Japanese Alocasia (A) odora. Materials and Methods: We investigated cases in Japan from our own hospital in the eastern part of Shizuoka Prefecture as well as published reports. Results: A. dorais found in western Japan, and plants of the Alocasia genus are cultivated often. A. odora is frequently associated with food poisoning because its aboveground parts resemble those of Satoimo (Colocasia esculenta). Moreover, A. odora contains insoluble calcium oxalate crystals, which cause poisoning symptoms, such as oral pain, nausea, vomiting, and laryngeal edema, resulting in near asphyxia, diarrhea following shock, and skin dermatitis. Calcium oxalate crystals are abundant in Araceae family plants, and cases of health damage owing to the accidental ingestion of Araceae plants have been reported worldwide. Conclusion: Due to the strong irritation felt in the mouth upon contact with the plant, it is advisable to immediately spit out the plant and rinse the mouth. In addition to drug administration, ensuring a secure airway may be necessary if there is a risk of asphyxiation.

Evaluation of Pharmacognostical and Phytochemical Studies for the Development of Quality Control Parameters for Stem Bark of Ximenia americana Linn. (Olacaceae)

Abstract Background: Plants are extremely useful to humankind as medicine, and hence the properties of medicinal plants have to be scientifically evaluated. Aim: The aim of this study is to establish pharmacognostical and phytochemical parameters for stem bark of Ximenia americana Linn. (Olacaceae). Materials and Methods: Macroscopical, microscopical evaluation, powder microscopical studies, quantitative microscopy, physicochemical evaluation, fluorescence analysis, analysis of inorganic elements, and determination of heavy metal contamination were performed as per the World Health Organization guidelines and standard methods. To evaluate various phytoconstituents, phytochemical investigations such as preliminary phytochemical screening, high-performance thin-layer chromatography, and high-performance liquid chromatography were performed. Results: Microscopical studies showed the presence of thick cortex, periderm cells, phloem parenchyma, phloem fibers, sieve elements, prismatic and granular calcium oxalate crystals, and resinous cells. The length and width of fibers, stone cells, and diameter of starch grains were measured. Physicochemical evaluation like ash values has shown total ash – 8.01% ± 0.02% w/w, acid-insoluble ash – 1.53% ± 0.02% w/w, water-soluble ash – 1.54% ± 0.01% w/w, sulfated ash – 12.02% ± 0.07% w/w, water-soluble hot extraction – 26.48 ± 0.02% w/w, cold maceration – 11.76% ± 0.04% w/w, alcohol soluble – 15.69% ± 0.03%w/w, ether soluble – 2.06% ± 0.03% w/w, and loss on drying – 4.88% ± 0.01% w/w. Conclusion: Pharmacognostical study on the stem bark of Ximenia americana may be reported as first. The results of pharmacognostical and phytochemical studies will be helpful for authentication and provide quality control parameters for the stem bark of this plant.

Screening of oxalate-degrading probiotics and preventive effect of Lactiplantibacillus plantarum AR1089 on kidney stones.

Calcium oxalate stone is the main type of kidney stone, so far there is no specific drug treatment. Here, we screened for oxalate-degrading probiotics and evaluated the potential preventive effect of lactic acid bacteria in rats with hyperoxaluria-induced kidney stones. The oxalate degradation efficiencies of the probiotics were determined to be 5-20% by in vitro experiments, of which the degradation efficiencies of Lactiplantibacillus plantarum AR342 and L. plantarum AR1089 were 17.32% and 14.15%, respectively. Through animal experiments, we found that L. plantarum AR1089 significantly attenuated kidney injury, as demonstrated by improving renal dysfunction and renal fibrosis, lowering creatinine and urea nitrogen levels. L. plantarum AR1089 was also effective in decreasing the number of calcium oxalate crystals in the urine and kidneys as well as ameliorating oxidative stress as evidenced by lowering the level of MDA and decreasing the level of SOD and CAT. Moreover, supplementation of L. plantarum AR1089 inhibited renal crystalline deposition by down-regulating the expression of KIM-1, OPN and MCP-1, and prevented hyperoxaluria-induced kidney stones by regulating the gut microbiota. Taken together, the present study shows that oral administration of L. plantarum AR1089, by attenuating kidney injury and regulating gut microbiota, is a potential therapy to reduce calcium oxalate crystals and prevent the progression of kidney stones.

Tratamento foliar com metil jasmonato na restrição do crescimento e na anatomia de foliar de Begônia ‘Dragon Wing’

Abstract The application of plant growth regulators, primarily gibberellin inhibitors, is the main approach used for controlling the growth of ornamental plants. However, there is a growing interest in alternative products due to the risk of environmental toxicity associated with traditional methods. Methyl jasmonate (MeJa) is a natural substance found in plants and is considered to have low toxicity when used appropriately. Its use as a growth regulator has shown promise for controlling plant growth, but its effects have not been extensively explored in ornamental growth restriction. This study investigated the effects of foliar application of different concentrations of MeJa (0, 50, 100, and 150 µM) on the growth and leaf anatomy of Begonia ‘Dragon Wing’ Pink and Red. Plant growth parameters were evaluated, and qualitative-quantitative histological analyses of Begonia leaves were conducted. MeJa was efficient in compacting the plants, reducing height in 17.57% and diameter in 8.98% without compromising biomass, root growth, and flowering aspects. The average concentration studied also caused changes in leaf anatomy, increasing the thickness of the abaxial epidermis, reducing mesophyll thickness, the number of calcium oxalate crystals, and the size and number of stomata. The results demonstrated the promising effect of MeJa on controlling the growth of potted ornamental plants.

PPARγ agonist alleviates calcium oxalate nephrolithiasis by regulating mitochondrial dynamics in renal tubular epithelial cell.

Kidney stone formation is a common disease that causes a significant threat to human health. The crystallization mechanism of calcium oxalate, the most common type of kidney stone, has been extensively researched, yet the damaging effects and mechanisms of calcium oxalate crystals on renal tubular epithelial cells remain incompletely elucidated. Regulated mitochondrial dynamics is essential for eukaryotic cells, but its role in the occurrence and progression of calcium oxalate (CaOx) nephrolithiasis is not yet understood. An animal model of calcium oxalate-related nephrolithiasis was established in adult male Sprague‒Dawley (SD) rats by continuously administering drinking water containing 1% ethylene glycol for 28 days. The impact of calcium oxalate crystals on mitochondrial dynamics and apoptosis in renal tubular epithelial cells was investigated using HK2 cells in vitro. Blood samples and bilateral kidney tissues were collected for histopathological evaluation and processed for tissue injury, inflammation, fibrosis, oxidative stress detection, and mitochondrial dynamics parameter analysis. Calcium oxalate crystals caused higher levels of mitochondrial fission and apoptosis in renal tubular epithelial cells both in vivo and in vitro. Administration of a PPARγ agonist significantly alleviated mitochondrial fission and apoptosis in renal tubular epithelial cells, and improved renal function, accompanied by reduced levels of oxidative stress, increased antioxidant enzyme expression, alleviation of inflammation, and reduced fibrosis in vivo. Our results indicated that increased mitochondrial fission in renal tubular epithelial cells is a critical component of kidney injury caused by calcium oxalate stones, leading to the accumulation of reactive oxygen species within the tissue and the subsequent initiation of apoptosis. Regulating mitochondrial dynamics represents a promising approach for calcium oxalate nephrolithiasis.

Polymer-Coated Urinary Catheter Reduces Biofilm Formation and Biomineralization: A First-in-Man, Prospective Pilot Study

Purpose: Biofilm formation and biomineralization on urinary catheters may cause severe complications including infection and obstruction. Here, we describe an in vitro evaluation and prospective pilot clinical study of a silicone catheter coated with a biofilm-resistant polymer. Materials and Methods: Biofilm biomass and biomineralization on uncoated and coated catheters were quantified by confocal microscopy using fluorescently tagged bacteria or stained for biofilm and minerals. Biomineral identity was determined using scanning electron microscopy and X-ray spectroscopy. Biofilm formation and biomineralization were evaluated in vitro using uropathogens Proteus mirabilis and Pseudomonas aeruginosa and on catheters recovered from hospitalized patients. Fibrinogen in patient urine and on catheters was quantified using an immunofluorescence assay. Results: In vitro P. mirabilis and P. aeruginosa formed significantly less biofilm and biomineral and failed to block coated compared with uncoated catheters in a bladder model after 89 h. Biofilm-resistant polymer-coated catheters (n = 83) recovered from hospitalized patients exhibited significantly lower biofilm biomass and biomineralization compared with uncoated silicone catheters (n = 78). Electron microscopy with elemental analysis of recovered catheters revealed calcium oxalate crystals on coated compared with the struvite and apatite crystals on uncoated catheters associated with catheter blockage. Lower levels of biofilm-promoting fibrinogen in postcatheterization urine and on catheters from patients receiving coated catheters was observed compared with those receiving uncoated catheters indicative of a reduced inflammatory response. Conclusions: These data provide evidence that polymer-coated urinary catheters exhibit enhanced resistance to fibrinogen deposition, biofilm formation, and encrustation, reducing the risks associated with catheter-associated urinary tract infections and obstruction.

Investigation of the potential anti-urolithiatic activity of Alhagi maurorum (Boiss.) grown wild in Al-Ahsa (Eastern Province), Saudi Arabia.

The potential of Alhagi maurorum (Boiss.) aqueous extract (AME), used in traditional medicine for treatment or prevention of urolithiasis, to dissolve calcium oxalate stones in vitro was evaluated. In order to determine the litholytic potential of the extract, Calcium oxalate urinary stones were incubated during 12 weeks under continuous shaking in the presence of AME, Rowanix or NaCl 9 g/mL solution were used as controls. After the incubation period, the residual weight of the treated calculi was determined and the rate of dissolution was calculated. The medium pH variation was measured and changes in the calcium oxalate crystals at the stone surface were assessed using a scanning electron microscope (SEM). The results showed a significant dissolution effect for the extract on the kidney calculi during the experimentation period. At the end of the experiment, the percentages of calculi weight decrease were 41.23, 4.97 and 55.67% for the extract, NaCl solution and Rowanix, respectively. Gas Chromatography analysis revealed mainly the presence of the following phyto-compounds: Cyclopropenone, 2,3-diphenyl; 1-Nonadecanol; methyl-alpha-D-mannopyranoside; cis-9-Hexadecenal. These compounds unarguably play crucial roles in the health care system especially in cancer treatment and many other diseases including urolithiasis. The urinary stone dissolution, independent of medium pH, could be attributed to formation of complexes between the phytochemical compounds in the extract and the calculi.

Effect of croton Tonkinensis Gagnep extract on calcium oxalate crystal formation causing kidney stones under in vitro conditions

The study investigated the use of Croton tonkinensis Gagnep extract to evaluate its ability to inhibit Calcium oxalate crystal formation, including three main stages: nucleation, growth, and aggregation. Croton tonkinensis Gagnep samples were extracted using a soaking method with 96% ethanol to obtain the concentrated extract. The percentage of inhibition of Calcium oxalate crystal nucleation by the Croton tonkinensis Gagnep extract was determined using spectrophotometry at a wavelength of 620 nm. Meanwhile, the inhibition efficacy on the growth of Calcium oxalate crystals was evaluated by measuring the optical density of the test sample at a wavelength of 214 nm over a period of 780 seconds. The inhibition efficacy on the aggregation of Calcium oxalate crystals by the extract was determined by measuring the optical density at a wavelength of 620 nm at different time intervals of 30, 60, 90, 180, 360, and 720 minutes. The research results showed that the extract had a high efficiency of 1.36%. The Croton tonkinensis Gagnep extract contained various compounds such as flavonoids, alkaloids, saponins, terpenoids, tannins, and phenols. The extract exhibited the ability to inhibit nucleation with an IC50 value of 7.78 mg/mL, as well as inhibit the growth and aggregation of Calcium oxalate crystals with times of 13 minutes and 90 minutes, respectively.

Hierarchical structure and chemical composition of complementary segments of the fruiting bodies of Fomes fomentarius fungi fine-tune the compressive properties.

Humanity is often fascinated by structures and materials developed by Nature. While structural materials such as wood have been widely studied, the structural and mechanical properties of fungi are still largely unknown. One of the structurally interesting fungi is the polypore Fomes fomentarius. The present study deals with the investigation of the light but robust fruiting body of F. fomentarius. The four segments of the fruiting body (crust, trama, hymenium, and mycelial core) were examined. The comprehensive analysis included structural, chemical, and mechanical characterization with particular attention to cell wall composition, such as chitin/chitosan and glucan content, degree of deacetylation, and distribution of trace elements. The hymenium exhibited the best mechanical properties even though having the highest porosity. Our results suggest that this outstanding strength is due to the high proportion of skeletal hyphae and the highest chitin/chitosan content in the cell wall, next to its honeycomb structure. In addition, an increased calcium content was found in the hymenium and crust, and the presence of calcium oxalate crystals was confirmed by SEM-EDX. Interestingly, layers with different densities as well as layers of varying calcium and potassium depletion were found in the crust. Our results show the importance of considering the different structural and compositional characteristics of the segments when developing fungal-inspired materials and products. Moreover, the porous yet robust structure of hymenium is a promising blueprint for the development of advanced smart materials.

Potassium oxalate-based mouth rinse for rapid relief in dentinal hypersensitivity

Abstract Dentinal hypersensitivity (DHS) affects about 1.3%–92.1% of the population globally and has a significant impact on the quality of life of the affected individuals. Desensitizing agents that directly block the dentinal tubules or cause chemical blocking of the pulpal nerve endings are commonly recommended for the management of DHS. Potassium oxalate (KO) has been used as a desensitizing agent in the management of DHS. This narrative review aimed to assess the literature related to the efficacy of KO as a desensitizing agent, with a focus on mouth rinses containing 1.4% KO. A comprehensive search of the literature was carried out using PubMed and Google Scholar with relevant keywords. The results from the retrieved articles were assessed, and the findings regarding the efficacy of KO in managing DHS were analyzed. Studies suggest that the usage of 1.4% KO mouth rinses results in the occlusion of dentinal tubules by the formation of acid-resistant calcium oxalate crystals within dentinal tubules and on the dentine surface. These calcium oxalate crystals tend to form a strong bond with the inner surface of the dentinal tubules and lower the hydraulic conductivity of the dentine. These actions of KO can ensure both rapid and long-term relief from DHS.

Anatomy of stem lesions caused by Citrus leprosis

Abstract The citrus leprosis is responsible for causing major damage in fruit production. The disease, in the stems, is characterized by caused “scaly bark” lesions of reddish tone, depending on the stage they can be called blisters and make ruptures in the longitudinal direction. The aim of this study was to analyze the evolution of leprosis lesions of type CiLV-C and the anatomical alterations in the twigs of field-collected plants in order to compare to the evolution of the foliar and twigs lesions of inoculated plants already described in literature. For anatomical analysis, the samples were fixed in Karnovsky solution, dehydrated in a graded ethylic series, embedded in hidroxy-ethyl methacrylate resin (Leica Historesin), sectioned (5-7 μm thick), and stained with Toluidine blue for usual histological analysis. Histochemical tests were also carried out. The digital images were capture in a microscope with video camera. The analyzed lesion followed the same pattern as described to foliar lesions, i.e., there was a necrotic center surrounded by a halo also necrotic. In the necrotic region there was the accumulation of lipid compounds. The cortical and, or phloematic parenchyma exhibited hyperplasia. In both lesions it was verified the decrease of starch amount and the increase of quantity of prismatic calcium oxalate crystals, and the presence of gummosis traumatic ducts.

Hydroalcoholic Extract of Ziziphus Jujuba Leaf to Prevent Ethylene Glycol and Ammonium Chloride-Induced Kidney Stones in Male Rat: Is it Effective?

This study aimed to evaluate the effect of Ziziphus jujuba (Z. jujuba) leaf hydroalcoholic extract on the prevention/treatment of kidney stones. Thirty-six male Wistar rats were randomly divided into six groups: control, Sham (kidney stone induction (KSI) by ethylene glycol 1% + ammonium chloride 0.25% through drinking water for 28 days), Prevention groups 1, 2 (KSI and Z. jujuba leaf (250 and 500 mg/kg, respectively) through gavage for 28 days), and Treatment groups 1, 2 (KSI and Z. jujuba leaf (250 and 500 mg/kg, respectively) from the 15th day). On the 29th day, the rats' 24-hour urine was assessed, the animals were weighed, and blood samples were taken. Finally, after nephrectomy and weighing the kidneys, tissue sections were prepared to examine the number of calcium oxalate crystals and tissue changes. The results indicated a significant increase in kidney weight and index, tissue changes, and the number of calcium oxalate crystals in the Sham group compared to the control; using Z. jujuba leaf considerably reduced them in experimental groups compared to the Sham. Body weight decreased in the Sham and experimental groups (except the prevention 2 group) compared to the control, while this observed reduction was lower in all experimental groups compared to the Sham. The mean urinary calcium, uric acid, creatinine, and serum creatinine in Sham and experimental groups (except the prevention 2 group) indicated a substantial increase compared to the control and decreased significantly in all experimental groups compared to the Sham. Hydroalcoholic extract of Z. jujuba leaf is effective in the reduction of calcium oxalate crystals forming, and its most effective dose was 500mg/kg.

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.