Sodium Phosphate Research Articles

Sustainable nutrient recovery through struvite precipitation from poultry and multi-substrate agricultural waste digestates

This study addresses the multifaceted challenges stemming from the excessive application of manure or digestate, with the aim of mitigating problems of storage, emissions, contamination and transportation. It presents a unique-protocol for efficiently recovering nutrients in the form of struvite, a pure, easy-to-use magnesium ammonium phosphate compound. The objective is to explore struvite production via mono-digestion of poultry manure and co-digestion of various organic wastes, including poultry, dairy, pig manures, and wasted corn silage. Crystals are synthesized under optimized conditions: magnesium/phosphorus ratio of 1.5, pH 9, and mixing at 240 rpm. The process achieves ≥95 % TAN-recovery in moisture-free form for various digestates, with a significant 61.5 % reduction in volatile-fatty-acids. Approximately 0.33–1 g struvite is produced per 10 mL digestate. The study suggests using disodium or monopotassium phosphate for practical implementation, demonstrating the versatility of digestates. Integrating this method into anaerobic digestion promotes the circular-economy and sustainable management of waste nutrients.

Comparing Conventional and Modern Methods for The Phycocyanin Extraction from Spirullina sp

Spirulina platensis, a blue-green algae abundant in tropical regions, is rich in minerals, vitamins, fibers, and pigments, with low nucleic acid content. It has unique chromoproteins called phycobiliproteins, notably phycocyanin, used in various applications. This study aims to optimize phycocyanin extraction using different solvents (distilled water and sodium phosphate buffer pH 6.7) and methods (freeze-thaw and sonication). Spirulina platensis biomass was extracted in both solvents, then some of them was freeze for 24 and 48 hours followed by thawing overnight. The other was sonicated for 2.5 minutes, 50 Hz then soaked for 1, 2, and 3 hours. All of the samples were centrifuged at 6000 rpm for 10 minutes and the absorbance was measured using a UV-Vis spectrophotometer at wavelengths of 280, 620, and 650 nm. with freeze-thawing for 48 hours yielded the highest phycocyanin concentration (0.55%), with a yield of 11.07 and purity of 0.21. Sonication improved phycocyanin concentration, yield, and purity significantly, yielding 1.108, 25.85, and 0.26, respectively.

Dexamethasone nanocrystals-embedded hydroxypropyl methylcellulose hydrogel increases cochlear delivery and attenuates hearing loss following intratympanic injection

Herein, dexamethasone (DEX) nanocrystalline suspension (NS)-embedded hydrogel (NS-G) was constructed using a hydroxypropyl methylcellulose (HPMC) polymer to enhance cochlear delivery and attenuate hearing loss following intratympanic (IT) injection. Hydrophobic steroidal nanocrystals were prepared using a bead milling technique and incorporated into a polysaccharide hydrogel. The NS-G system with HPMC (average molecular weight, 86,000 g/mol; 15 mg/mL) was characterized as follows: rod-shaped drug crystalline; particle size <300 nm; and constant complex viscosity ≤1.17 Pa·s. Pulverization of the drug particles into submicron diameters enhanced drug dissolution, while the HPMC matrix increased the residence time in the middle ear cavity, exhibiting a controlled release profile. The IT NS-G system elicited markedly enhanced and prolonged drug delivery (> 9 h) to the cochlear tissue compared with that of DEX sodium phosphate (DEX-SP), a water-soluble prodrug. In mice with kanamycin- and furosemide-induced ototoxicity, NS-G markedly enhanced hearing preservation across all frequencies (8–32 kHz), as revealed by an auditory brainstem response test, compared with both saline and DEX-SP. Moreover, treatment with NS-G showed enhanced anti-inflammatory effects, as evidenced by decreased levels of inflammation-related cytokines. Therefore, the IT administration of DEX NS-loaded HPMC hydrogels is a promising strategy for treating hearing loss.

Influence of NaCl and phosphate on gelation properties of chicken breast myofibrillar protein gels and its application to in vitro digestion model

To investigate the combination effect of sodium chloride and phosphates on chicken breast myofibrillar proteins, MP gels containing various molarity of NaCl (0.15, 0.30 and 0.45 M) and phosphate (0 and 0.05 M) were prepared, their rheological properties were characterized, and applied to an in vitro digestion model. MP mixture containing 0.45 M NaCl and 0.05 M phosphate had the highest viscosity. The gel strength and cooking yield of MP gels was improved by increasing of molarity of NaCl. As NaCl concentration in MP increased, sulfhydryl levels decreased, while disulfide levels increased. As NaCl and phosphate levels increase, MP gels become denser and porosity decreases, which may reduce protein digestibility. In SDS-PAGE, protein bands from MP gels containing low NaCl levels (≤ 0.30 M) degraded more rapidly during in vitro digestion. These results may support the need for the meat industry to develop low-salt meat products with improved digestibility. KeywordsChicken,Myofibrillar protein,NaCl,Phosphate,Rheological properties,In vitro digestion

A Stereolithography-Based Modified Spin-Casting Method for Faster Laboratory-Scale Production of Dexamethasone-Containing Dissolving Microneedle Arrays.

Microneedle arrays (MNAs) consist of a few dozens of submillimeter needles, which tend to penetrate through the stratum corneum layer of the skin and deliver hardly penetrating drugs to the systemic circulation. The application of this smart dosage form shows several advantages, such as simple use and negligible pain caused by needle punctures compared to conventional subcutaneous injections. Dissolving MNAs (DMNAs) represent a promising form of cutaneous drug delivery due to their high drug content, biocompatibility, and ease of use. Although different technologies are suitable to produce microneedle arrays (e.g., micromilling, chemical etching, laser ablation etc.), many of these are expensive or hardly accessible. Following the exponential growth of the 3D-printing industry in the last decade, high-resolution desktop printers became accessible for researchers to easily and cost-effectively design and produce microstructures, including MNAs. In this work, a low force stereolithography (LFS) 3D-printer was used to develop the dimensionally correct MNA masters for the spin-casting method. The present study aimed to develop and characterize drug-loaded DMNAs using a two-level, full factorial design for three factors focusing on the optimization of DMNA production and adequate drug content. For the preparation of DMNAs, carboxymethylcellulose and trehalose were used in certain amounts as matrices for dexamethasone sodium phosphate (DEX). Investigation of the produced DexDMNAs included mechanical analysis via texture analyzer and optical microscopy, determination of drug content and distribution with HPLC and Raman microscopy, dissolution studies via HPLC, and ex vivo qualitative permeation studies by Raman mapping. It can be concluded that a DEX-containing, mechanically stable, biodegradable DexDMNA system was successfully developed in two dosage strengths, of which both efficiently delivered the drug to the lower layers (dermis) of human skin. Moreover, the ex vivo skin penetration results support that the application of DMNAs for cutaneous drug delivery can be more effective than that of a conventional dermal gel.

RP-HPLC Method Development and Validation for Simultaneous Determination of Sofosbuvir and Ledipasvir: A Pharmaceutical Application Study

The objective of the present work was to develop a simple, precise, accurate, reproducible, and specific reversed-phase high performance liquid chromatographic method for the simultaneous determination of sofosbuvir and ledipasvir in pharmaceutical dosage form. The HPLC system (Shimadzu, LC-2030) consisted of Hypersil ODS - C18 (250 mm, 4.6 mm, 5 μm- Thermo Scientific) was used for the isocratic elution using mobile phase composed of sodium phosphate buffer (pH 3.5) and acetonitrile in the ratio of 40:60% (v/v). Both sofosbuvir and ledipasvir were detected at 260 nm with a flow rate of 1 mL/min. The mean retention time of sofosbuvir was 3.094, whereas ledipasvir was 6.22 minutes. A linear relationship was found in the concentration ranged from 0.54 to 162 μg/mL for sofosbuvir (r2 = 0.9993) and 0.14 to 42 μg/mL for ledipasvir (r2 = 0.9994). The RSD values for recovery test were found 0.99% for sofosbuvir and 0.22% for ledipasvir. The lower limit of detection (LOD) were 0.054 mcg/mL and 0.012 mcg/mL, and the limit of quantification (LOQ) were 0.135 mcg/mL and 0.030 mcg/mL for sofosbuvir and ledipasvir, respectively. The method was successfully applied to the assay and in-vitro dissolution studies of sofosbuvir and ledipasvir in tablet dosage form and all the results were found within the acceptance limits.

Influence of phosphates on phase formation in alkali‐activated MgO‐Al2O3‐SiO2‐P2O5 cements

AbstractThe impact of phosphates on phase formation in low Ca alkali‐activated materials (AAMs) is investigated using a polymer‐assisted sol‐gel process to fabricate MgO‐Al2O3‐SiO2‐P2O5 cement precursors covering a broad range of compositions activated with sodium hydroxide. X‐ray diffraction and magic angle spinning‐nuclear magnetic resonance are used to examine the crystalline and amorphous phases that form over 470 days of curing (35°C, 90% relative humidity). The results confirm that Al is preferentially incorporated into hydrotalcite‐like layered double hydroxides (LDH) over zeolites. Zeolites form when more Al is present than can be incorporated into the LDH. Little evidence of phosphate incorporation into aluminosilicate networks (such as zeolites or disordered aluminosilicate hydrate) was observed. The phosphates present in the precursor favor reaction with sodium to form water‐soluble sodium phosphate phases. In most cases, the remainder of the phosphates become adsorbed to the surface of other phases and are not intercalated into the LDH, though at high phosphate concentrations (26.6 wt. % P2O5) and extended curing times (470 days), phosphates are observed to intercalate into LDH phases. These results provide preliminary evidence that phosphates are compatible with low Ca AAMs, which is consequential as there is a growing interest in both the use of AAM and phosphate‐based corrosion inhibiter in steel‐reinforced concrete.

Graphene nanoplatelets-infused binary eutectic phase change materials for enhanced thermal energy storage

Energy depletion for the thermal regulation of buildings is a major global concern. Herein, we develop a binary eutectic phase change material (EPCM) consisting of sodium sulphate decahydrate (SSD) and sodium phosphate dibasic dodecahydrate (SPDD) that were modified using borax, carboxymethyl cellulose (CMC), and graphene nanoplatelets (GNP). In the EPCM, SSD, and SPDD were blended at a proportion of 62:38 to obtain a eutectic point of 27.8 °C with a melting enthalpy of 202 J/g. Borax was a nucleating additive to lessen the issue of supercooling, CMC was a thickening agent to ensure phase stability, and GNP was added as a nanomaterial to enhance the thermal behavior of EPCM. Microtopography, chemical stability, optical transmissibility and absorptivity, phase transition characteristic, thermal conductivity, energy storage potential, degree of supercooling, and thermal stability over 200 thermal cycles were tested and determined. Using scanning electron microscopy, the GNP microstructure and its interaction with EPCM are visualized to present the dispersion and formation of the thermal network. The nanocomposite EPCM also has chemical stability, which is aided by a 571% increase in optical absorbance and an 83.8% decrease in transmissibility. With GNP as a nanomaterial, owing to the well-developed thermal network, we achieve a 106% increment in thermal conductance from 0.464 W/m⋅K to 0.956 W/m⋅K; likewise, GNP acts as a conducting binder and enhances the intermolecular interaction leading to an increase in melting enthalpy from 202.4 J/g to 226 J/g. Finally, a numerical simulation of EPCM heating a thermic fluid flowing within a concentric tube with focus on the temperature contour is conducted for elaborating the potential of PCM towards TES systems.

Observational Clinical Investigation Evaluating an Ophthalmic Solution Containing Xanthan Gum and Low Concentration Desonide Phosphate in Dry Eye Disease Treatment.

Patients with dry eye disease (DED) complain of a multitude of symptoms that affect their visual function and quality of life (QoL). This clinical investigation assessed the performance, tolerance, and safety of a novel preservative-free ophthalmic solution containing xanthan gum 0.2% and desonide sodium phosphate 0.025%. This was an observational, prospective, multicentric, and post-market clinical investigation to assess the effect of three times a day instillation of the study formulation in patients suffering from DED. The primary objective was to achieve a 50% reduction in conjunctival hyperemia index as assessed with the OCULUS Keratograph after 1 month of treatment compared to baseline values. The secondary objectives included patient-reported outcomes, clinical performance, and safety. Thirty patients were enrolled (21 women, 9 men) with a mean age of 61.10 ± 14.53years. The instillation of the study formulation was associated with a significant reduction in redness scores after 1 month of treatment compared to baseline (mean - 0.51 ± 0.51; p ≤ 0.0001). Although the primary endpoint was not completely met, a 50% reduction in the conjunctival hyperemia index was achieved in 23% of the participants, and 77% showed a reduction of 26% of the same index. In addition, the ophthalmic solution significantly increased tear film break-up time, and a significant reduction of corneal and conjunctival staining with fluorescein was achieved. It also reduced DED symptoms and had a very good safety profile. the study formulation produced a significant improvement in the signs, symptoms, and QoL of patients with mild to moderate DED with a good safety profile after 1 month of treatment.

Direct Screening of PET Hydrolase Activity in Culture Medium Based on Turbidity Reduction.

The development of an efficient screening method for the activity of PET-degrading enzymes represents a significant technological advance in the field of enzyme research, with the potential to facilitate the advancement of enzymes for PET recycling. By examining the stable conditions of PET suspension and enzyme production conditions, we developed a method to quantify PET-degrading enzyme activity in E. coli culture medium using turbidity reduction as an indicator. High PET concentration or ionic strength caused aggregation of PET, and the best condition for activity detection was 0.5 mg mL-1 PET in 50 mM sodium phosphate pH 7.0. Preculture of E. coli increased the purity of enzyme secreted in medium. To evaluate the screening method, 720 colonies of the PET2-7M-H229X-F233X mutant library were analyzed and three candidates of high-activity mutants were obtained. The thermostability of the mutants could also be easily measured by measuring the residual activity after heat treatment. The H229T-F233M mutant showed 3.4 times higher degradation rate against PET film than the template enzyme at the initial time. The molecular dynamics simulation implied that the F233M mutation makes space for making an α helix and that the H229T mutation resolved the steric hindrance with Trp199. These mutations were speculated to change the angle of the Trp199 side chain of PET2 to an angle similar to that of the Trp185 of IsPETase, making it suitable for PET binding to the active center. Screening of activity using PET suspensions is compatible with robotic automation and is expected to be useful for validating computationally predicted mutations.

Synthesis of hydroxyapatite and strontium-substituted hydroxyapatite for bone replacement and osteoporosis treatment

Hydroxyapatite (HAp) is an inorganic component exhibiting bioactivity similar to that of natural bone. However, it is not resorbed by osteoclasts during bone remodelling due to its lack of bio-resorption property. This can be enhanced by the substitution of other element presented in bone mineral. In this research work, hydroxyapatite (HAp) and strontium-substituted hydroxyapatite (Sr-HAp) were synthesized by a precipitation method. Calcium nitrate tetra hydrate [Ca(NO3)2•4H2O], disodium hydrogen phosphate (Na2HPO4), and Strontium nitrate [Sr(NO3)2] were used as Ca, PO4 and Sr sources, respectively. Molar ratio Ca/P=1.67 was used to synthesize HAp, where (Ca+Sr)/P=1.67 was used to synthesize strontium substituted-HAp (Sr-HAp). The reaction was carried out at room temperature. The results show that pure HAp and Sr-HAp were formed with nanometer-sized particles. Sr substitution in the HAp lattice results in an increase in both the lattice disorder and crystal aspect ratio. The results of in vitro bioactive testing using simulated bodily fluid also showed that both HAp and Sr-HAp have high bioactive, with the Sr-HAp sample having the greater bioactive. Therefore, HAp and Sr-HAp have great potential for biological applications.

DOE-based process optimization for development of efficient methods for purification of recombinant hepatitis B surface antigen from Pichia pastoris feedstock using Capto adhere resin

BackgroundThe multimodal chromatography resins, such as Capto adhere, are considered good candidates to be utilized in downstream processing due to their high capacity and selectivity; however, their multimodal interactions lead to an intricacy in the adsorption-desorption patterns and systematic characterization of conditions for process steps is necessary. MethodsCapto adhere, a strong ion exchanger with multimodal functionality, was used in this study for the final aim of recombinant hepatitis B surface antigen (rHBsAg) purification from Pichia pastoris (P. pastoris) industrial feedstock. Optimization of various parameters was done using the design of experiments (DOE) approach to determine the best binding and non-binding conditions. ResultsMaximum rHBsAg binding on Capto adhere occurred in 20 mM sodium acetate, pH 4.5, and a binding capacity of about 0.75 mg/ml was achieved, which was much higher than rHBsAg binding capacity of other resins reported so far. In elution optimization investigations, it was revealed that 1 M arginine (buffered in 50 mM sodium phosphate, pH 6.5) was the most efficient eluting agent. The binding and elution optimal conditions were utilized for further purification of rHBsAg from P. pastoris industrial feedstock in bind-elute mode, and the recovery and purity of the obtained rHBsAg were about 60% and 100%, respectively. Following optimization in the flow-through purification mode, the target protein recovery was significantly increased (up to 97%) and the target protein purity of more than 95% was achievable. SEC-HPLC analysis showed that the obtained retention times for the purified rHBsAg were similar to those reported previously. ConclusionsThese results suggest that Capto adhere under such optimized conditions can be considered as a good candidate for efficient purification of rHBsAg from P. pastoris industrial feedstock in downstream processing.

Elucidating the roles of electrolytes and hydrogen bonding in the dewetting dynamics of the tear film

This study explores the impact of electrostatic interactions and hydrogen bonding on tear film stability, a crucial factor for ocular surface health. While mucosal and meibomian layers have been extensively studied, the role of electrolytes in the aqueous phase remains unclear. Dry eye syndrome, characterized by insufficient tear quantity or quality, is associated with hyperosmolality, making electrolyte composition an important factor that might impact tear stability. Using a model buffer solution on a silica glass dome, we simulated physiologically relevant tear film conditions. Sodium chloride alone induced premature dewetting through salt crystal nucleation. In contrast, trace amounts of solutes with hydroxyl groups (sodium phosphate dibasic, potassium phosphate monobasic, and glucose) exhibited intriguing phenomena: quasi-stable films, solutal Marangoni-driven fluid influx increasing film thickness, and viscous fingering due to Saffman-Taylor instability. These observations are rationalized by the association of salt solutions with increased surface tension and the propensity of hydroxyl-group-containing solutes to engage in significant hydrogen bonding, altering local viscosity. This creates a viscosity contrast between the bulk buffer solution and the film region. Moreover, these solutes shield the glass dome, counteracting sodium chloride crystallization. These insights not only advance our understanding of tear film mechanics but also pave the way for predictive diagnostics in dry eye syndrome, offering a robust platform for personalized medical interventions based on individual tear film composition.

Study on the mechanism of Na2CO3-roasting decomposition for water leach residue

In the process of treating cerium fluorocarbon-cerium lanthanide mixed rare earth concentrates by sulfuric acid roasting method, a large amount of waste leach residue containing iron, rare earths and phosphorus produced by flood neutralization needs to be solved urgently. In this paper, sodium carbonate roasting decomposition was used to treat the water leach residue, in which iron and rare earths were transformed into oxides, and the phosphorus was transformed into sodium phosphate. The main reactions and thermodynamic mechanisms of the roasting decomposition process were investigated by thermogravimetric analysis, phase analysis and chemical analysis. When the mass ratio of sodium carbonate to water leach residue is 1.5:1, the roasting temperature is 700 °C, and the roasting time is 1.5 h, the leaching rate of phosphorus with the roasted product reaches more than 98%. Meanwhile, the phase of the roasted product after washing mainly consists of iron oxide and rare earth oxides. The combination of sodium carbonate roasting decomposition and water leaching is effective for the treatment of water leach residue, which provides an experimental and theoretical basis for solving the problem of environmental and resource waste caused by the accumulation of a large amount of water leach residue. In addition, because sodium carbonate can achieve the separation of iron and phosphorus, this method also has certain reference value for the recovery and utilization of iron phosphate in lithium iron phosphate battery waste.

Enhancing water resistance in magnesium oxychloride cement modified with sodium dihydrogen phosphate at low MgO concentrations

Phosphate ions significantly enhance the water resistance of magnesium oxychloride cement (MOC). In this study, MOC was prepared with a MgO/MgCl2 molar ratio of 7 and a H2O/MgCl2 molar ratio of 12, while varying the dosage of sodium dihydrogen phosphate. The aim was to observe the microstructural changes in 5Mg(OH)2·MgCl2·8H2O (Phase 5) crystals and the macroscopic mechanical properties before and after immersion. The research found that phosphate ions increase the content of air-cured Phase 5 and reduce the formation of Mg(OH)2 by lowering the required Mg2+ concentration for Phase 5 production. However, excessive phosphate ions can damage the crystal structure of Phase 5, leading to a decrease in MOC strength. An appropriate amount of phosphate ions increases the output of Phase 5, resulting in a highly dense microstructure, which significantly improves the water resistance of MOC. Additionally, this enhances the pore structure of MOC, making the overall structure more compact and resistant to water penetration. This research offers valuable theoretical insights into Phase 5 crystal variations during air curing and water-immersion damage under low Mg2+ concentrations.

Silica-based aerogels encapsulate organic/inorganic composite phase change materials for building thermal management

Phase change energy storage materials are widely used in thermal management because they reduce energy consumption and effectively address issues such as energy supply and demand mismatches. However, challenges such as leakage in solid-liquid phase change materials (PCMs), flammability in organic solid-liquid PCMs, and supercooling and phase separation in inorganic solid-liquid PCMs still need to be resolved for their practical applications. In this study, we used methyl palmitate, dodecahydrate disodium hydrogen phosphate, and decahydrate sodium carbonate as the main PCMs, sodium carboxymethyl cellulose as an emulsifier and thickener, and silica aerogel as the encapsulating material to successfully prepare a novel shape-stabilized organic/inorganic composite PCM (CPCM). The prepared CPCM exhibits excellent shape stability and flame retardancy without supercooling or phase separation issues. In addition, the prepared CPCM has a high latent heat value (174.1 J/g) and a suitable phase transition temperature (22.9 °C), and its thermal performance remains basically unchanged after 100 heating/cooling cycles. Furthermore, this CPCM also demonstrates exceptional performance in building thermal management. This study provides a new solution to the problems of flammability, supercooling, phase separation and easy leakage of CPCM currently used in the field of building thermal management.

Extraction and Structural Analysis of Sweet Potato Pectin and Characterization of Its Gel.

Pectin is widely used in the food and pharmaceutical industries. However, data on sweet potato pectin extraction and structural property analyses are lacking. Here, for the high-value utilization of agricultural processing waste, sweet potato residue, a byproduct of sweet potato starch processing, was used as raw material. Ammonium oxalate, trisodium citrate, disodium hydrogen phosphate, hydrochloric acid and citric acid were used as extractants for the pectin constituents, among which ammonium oxalate had a high extraction rate of sweet potato pectin, low ash content and high molecular weight. Structural and gelation analyses were conducted on ammonium oxalate-extracted purified sweet potato pectin (AMOP). Analyses showed that AMOP is a rhamnogalacturonan-I-type pectin, with a molecular weight of 192.5 kg/mol. Chemical titration and infrared spectroscopy analysis confirmed that AMOP is a low-ester pectin, and scanning electron and atomic force microscopy demonstrated its linear molecular structure. Gelation studies have revealed that Ca2+ is the key factor for gel formation, and that sucrose significantly enhanced gel hardness. The highest AMOP gel hardness was observed at pH 4, with a Ca2+ concentration of 30 mg/g, pectin concentration of 2%, and sucrose concentration of 40%, reaching 128.87 g. These results provide a foundation for sweet potato pectin production and applications.

Defect-Engineered Metal-Organic Frameworks as Nanocarriers for Pharmacotherapy: Insights into Intracellular Dynamics at The Single Particle Level.

Nanoscale Metal-Organic Frameworks (nanoMOFs) are widely implemented in a host of assays involving drug delivery, biosensing catalysis, and bioimaging. However, the cell pathways and cell fate remain poorly understood. Here, a new fluorescent nanoMOF integrating ATTO 655 into surface defects of colloidal UiO-66 is synthesized, allowing to track the spatiotemporal localization of Single nanoMOF in live cells. Density functional theory reveals the stronger binding of ATTO 655 to the Zr6 cluster nodes compared with phosphate and Alendronate Sodium. Parallelized tracking of the spatiotemporal localization of thousands of nanoMOFs and analysis using machine learning platforms reveals whether nanoMOFs remain outside as well as their cellular internalization pathways. To quantitatively assess their colocalization with endo/lysosomal compartments, a colocalization proxy approach relying on the nanoMOF detection of particles in one channel to the signal in the corresponding endo/lysosomal compartments channel, considering signal versus local background intensity ratio and signal-to-noise ratio is developed. This strategy mitigates colocalization value inflation from high or low signal expression in endo/lysosomal compartments. The results accurately measure the nanoMOFs' colocalization from early to late endosomes and lysosomes and emphasize the importance of understanding their intracellular dynamics based on single-particle tracking for optimal and safe drug delivery.

Determination of the Nucleic Acid Constituents in Edible Mushrooms by High-Performance Liquid Chromatography with Diode Array Detection (HPLC-DAD)

Nucleosides and nucleobases in edible mushrooms collected from the Tekirdağ region of Turkey were determined by high performance liquid chromatography (HPLC). Separation of compounds was accomplished by gradient elution on a C18 column. A mixture of methanol (MeOH):50 mM sodium phosphate buffer (1:99, v/v) at pH 3.70 and methanol:50 mM sodium phosphate buffer (20:80) at pH 6.20 was used as the solvent. Two methods were used to treat the lyophilized mushrooms: ultrasound-assisted solid-liquid extraction (US-SLE) and pressurized (accelerated) solid-liquid extraction (PSLE). Deionized water was used as the extraction solvent. The extraction temperature and time were optimized. Fourteen nucleosides and nucleobases were simultaneously monitored at 254 nm within 25 min. Ultrasound-assisted solid-liquid was demonstrated to be more effective, perhaps because the pressurized liquid extraction system is a closed system and the matrix-solvent interaction may be insufficient. Adenosine, guanosine and uridine were the most abundant compounds in all studied mushrooms.

Fluoride induces immunotoxicity by regulating riboflavin transport and metabolism partly through IL-17A in the spleen

The impairment of the immune system by fluoride is a public health concern worldwide, yet the underlying mechanism is unclear. Both riboflavin and IL-17A are closely related to immune function and regulate the testicular toxicity of fluoride. However, whether riboflavin or IL-17A is involved in fluoride-induced immunotoxicity is unknown. Here, we first established a male ICR mouse model by treating mice with sodium fluoride (NaF) (100 mg/L) via the drinking water for 91 days. The results showed that fluoride increased the expression of the proinflammatory factors IL-1β and IL-17A, which led to splenic inflammation and morphological injury. Moreover, the expression levels of the riboflavin transporters SLC52A2 and SLC52A3; the transformation-related enzymes RFK and FLAD1; and the key mitochondrial functional determinants SDH, COX, and ATP in the spleen were measured via real-time PCR, Western blotting, and ELISA. The results revealed that fluoride disrupted riboflavin transport, transformation, metabolism, and mitochondrial function. Furthermore, wild-type (WT) and IL-17A knockout (IL-17A-/-) C57BL/6 J male mice of the same age were treated with NaF (24 mg/kg·bw, equivalent to 100 mg/L) and/or riboflavin sodium phosphate (5 mg/kg·bw) via gavage for 91 days. Similar parameters were evaluated as above. The results confirmed that fluoride increased riboflavin metabolism through RFK but not through FLAD1. Fluoride also affected mitochondrial function and activated neutrophils (marked with Ly6g) and macrophages (marked with CD68) in the spleen. Interestingly, IL-17A partly mediated fluoride-induced riboflavin metabolism disorder and immunotoxicity in the spleen. This work not only reveals a novel toxic mechanism for fluoride but also provides new clues for exploring the physiological function of riboflavin and for diagnosing and treating the toxic effects of fluoride in the environment.