The widespread application of detonation nanodiamonds (DNDs) is limited by surface-coated non-diamond sp2 carbon impurities. In this work, an efficient salt-assisted catalytic purification strategy is developed to achieve selective oxidation removal of sp2 carbon. DND black powder was mixed with various chloride, carbonate, and bicarbonate salts and thermally treated in air to systematically investigate the effects of anions and cations on purification efficiency. Thermogravimetric analysis reveals that all tested salts significantly reduce the oxidation onset temperature of sp2 carbon and exhibit distinct catalytic trends: for anions, bicarbonates > carbonates > chlorides; for cations, Cs+ ≈ K+ > Na+. Among them, KHCO3 introduced via a wet-wrapping method shows the optimal performance, lowering the oxidation temperature by approximately 160 °C. Moreover, the wet-wrapping process effectively suppresses particle sintering and agglomeration during purification, resulting in purified DNDs with reduced average particle size and markedly improved dispersibility. Mechanistic investigations demonstrate that free alkali metal cations act as active sites, preferentially catalyzing sp2 carbon oxidation through a synergistic oxygen spillover-electron transfer mechanism. This study provides an effective and highly selective approach for DND purification. The proposed salt-assisted strategy, integrating catalytic oxidation and dispersion control, also offers valuable insights for the preparation of high-performance nanomaterials.
Previous studies have demonstrated that C. camphora exhibits diverse bioactivities, but its roots, particularly their phenolic constituents, remain largely unexplored. In this study, we optimized the extraction and purification of total phenolics from C. camphora roots, isolated their chemical constituents, and evaluated their bioactivities. Response surface methodology yielded optimal conditions of 71% ethanol, 78 °C, and a liquid-to-solid ratio of 24:1 (mL/g), giving a total phenolic content of 3.60 mg/g. Purification with HPD-600 resin followed pseudo-second-order kinetics (R2 = 0.9987). From the enriched phenolic fraction, twelve phenolic compounds were isolated, seven of which are reported from C. camphora roots for the first time. The enriched fraction exhibited strong antioxidant activities (DPPH IC50 = 107.21 μg/mL; •OH IC50 = 130.7 μg/mL; O2- IC50 = 141.70 μg/mL) and significant pancreatic lipase inhibitory activity (IC50 = 0.80 mg/mL). This integrated approach expands the chemical diversity of C. camphora roots and highlights their potential as a natural source of antioxidants and lipid-lowering agents.
Fibrous hydrophilic nanocarbon (HNC) materials were synthesized via the catalytic reaction of a gas mixture of carbon monoxide (CO) and hydrogen (H2), i.e., syngas. The synthesized HNC features high specific surface areas and strong adsorption capabilities, rendering it effective for water purification. The HNC surface consists of graphite edge planes functionalized with phenolic hydroxyl groups through interactions with a synthetic byproduct, H2O vapor. The expanded interlayer spacing of the functionalized edge planes offers an open framework rich in adsorption sites for water-soluble chemical species. Adsorption tests with aqueous dye solutions and actual wastewater demonstrated that HNC outperforms conventional activated carbon, owing to the ability to form strong hydrogen bonds between surface phenolic groups and contaminants.
The development of advanced porous functional materials with enhanced photocatalytic performance for the degradation of hazardous pollutants such as dyes has led to the emergence of graphene-/graphitic carbon nitride-based metal-organic frameworks (GBMOFs) as promising photocatalysts. This study aims to comprehensively review advances in GBMOFs for the photocatalytic degradation of dye pollutants. As part of the study objective, the photocatalytic performance, recyclability, and stability dynamics of GBMOFs were critically reviewed to identify patterns and key trends. Emerging trends and technology readiness levels were succinctly discussed to assess the industrial applicability of GBMOF photocatalytic systems for dye remediation. Interestingly, the findings revealed that most GBMOF systems can deliver >80% degradation efficiency at pH 3-13 and doses of 2-1000 mg in 3-390 minutes. A predominant operational trend involving ˙OH and ˙O2 - was also noticed. Furthermore, the study shows that GBMOFs can be recycled 3-30 times while maintaining >70% of their original degradation performance and morphology in most of the cases. Recent advances and emerging trends include the application of p-n-p heterojunctions, ternary GBMOFs (which outperformed binary counterparts), piezo and sono-assisted photocatalysis, the use of natural sunlight, oxygen vacancies, and defect engineering. Finally, future perspectives are presented to establish a clear framework for guiding the future design of highly efficient, stable, practical, and enhanced GBMOF photocatalytic systems for the remediation of various dyes and pollutants.
L-theanine is a characteristic non-proteinogenic amino acid found in tea leaves and has attracted considerable attention because of its diverse physiological activity and broad application prospects. γ-glutamyl transpeptidase (GGT) can catalyze the synthesis of L-theanine from L-glutamine and ethylamine without ATP consumption, highlighting its advantages for enzymatic production. In this study, a complete process was established for L-theanine production. Through screening of single and dual promoters, the optimal expression combination, PyxiE-PspoVG, was identified. Furthermore, by integrating the dal selection marker, an antibiotic-free engineered strain was developed. After flask-level optimization, the GGT activity reached 27.32 U/mL and further increased to 127.37 U/mL in 3 L fed-batch fermentation. Using the fermentation broth as the biocatalyst, fed-batch conversion of 0.6 M L-glutamine and 2 M ethylamine yielded 0.52 M (91.44 g/L) L-theanine within 24 h. Further integration of ceramic membrane filtration, ultrafiltration, nanofiltration, electrodialysis, activated-carbon decolorization and ethanol crystallization afforded a final product purity of 95.6%. This study offers a useful reference for large-scale L-theanine production.
Metal-organic framework (MOF)-fiber composites integrate MOF's tunable functionality with flexible fibrous matrices for applications in various areas, including water purification, filtration, gas adsorption, and sensing to catalysis. Conventional techniques for the fabrication of MOF-fiber composites often involve the use of harsh organic solvents and complex time-intensive processing steps and can result in composites with poor MOF adhesion. In our work, we address these challenges by developing a simple and solvent-free air-spray coating method for the fabrication of MOF-fiber web composites. We employ bicomponent nonwovens as our fibrous substrate, consisting of a shell/core structure with polypropylene in the core and poly(ether-block-amide) in the shell. MOF particles, zeolitic imidazolate framework-8 (ZIF-8) and UiO-66-NH2, are spray-coated onto a nonwoven fabric using compressed air in <1 min, followed by thermal treatment. This results in strong adhesion of MOF particles in the nonwoven matrix as well as fiber fusion, which enhances the mechanical strength of the resulting composites. Fabrication time (<25 min) is significantly lower than conventional methods. By optimizing the shell/core ratio in the fibers and thermal treatment conditions, we achieve surface areas of 66 m2/g for ZIF-8 and 162 m2/g for UiO-66-NH2 composites, respectively. Finally, ZIF-8 fiber web composite demonstrates >99% removal efficiency of copper ions (Cu(II)) from aqueous solutions, underscoring the applicability for water purification. Beyond ZIF-8 and UiO-66-NH2, the air-spray coating method is adaptable for other mechanically and thermally stable MOFs and various fibrous and nanofibrous structures. Overall, this work presents a scalable, simple, and solvent-free approach for MOF-fiber composite synthesis with broad applicability.
Glucagon-like peptide-1 (GLP-1), a 31-amino acid incretin hormone, is widely used in the treatment of type 2 diabetes mellitus due to its glucose-dependent insulinotropic activity. However, its small size makes it highly prone to proteolytic degradation in microbial expression systems such as Escherichia coli, leading to reduced manufacturing yield. While fusion to cleavable protein tags can improve peptide stability during purification, excessively large tags often compromise the overall yield, especially when the target peptide is smaller than the fusion partner. To overcome this limitation, we have engineered 11 cleavable fusion tag constructs (LP1-LP11) for recombinant expression of Arg34-GLP-1(7-37) (liraglutide precursor) in E. coli. The 11 constructs differed only in the tags. The expression vector contained a T7 leader sequence, affinity tags (6×His/6×Arg), inclusion body tags (11-125 amino acids), and TEV protease cleavage sites. Among the 11 tags, LP8 with a compact 4.0 kDa tag achieved the highest expression, yielding 133 mg/L of fusion protein and a calculated liraglutide precursor yield of 60 mg/L based on mass fraction (45% of fusion mass), with an actual recovered yield of 14.6 mg/L after RP-HPLC purification, largely due to efficient inclusion body formation (>95% insolubility) and enhanced translational initiation driven by the T7 leader sequence. The purified peptide's identity and sequence integrity were confirmed by LC/MS analysis. The primary advantage of this approach is mass fraction optimization which focuses on minimizing fusion-tag mass to maximize yield relative to the tag size without compromising inclusion-body formation thereby providing a scalable and economical approach for GLP-1 analogs and potentially other peptide-based biopharmaceuticals.
Recent clinical trials have highlighted the potential of P63+ lung progenitor cell (LPC) transplantation for lung repair and regeneration. Currently, ex vivo P63+ LPC expansion depends on coculture with growth-arrested fibroblast feeder cells (GAFs), necessitating repeated purification during passaging. While differential enzymatic digestion (DED) and fluorescence- or magnetic-activated cell sorting techniques (FACS/MACS) offer partial solutions, a scalable, efficient, and consistent separation technique remains unmet, particularly for cell therapy manufacturing. Here, we present a multidimensional double spiral (MDDS) inertial microfluidic device designed for high-throughput label-free enrichment of P63+ LPCs. The MDDS device achieves cell size-based separation of P63+ LPCs and growth-arrested feeder cells, processing at a rate of 106 to 107 cells per minute. MDDS-sorted P63+ LPC purity correlates with the LPC-to-GAF ratio in culture. With an initial ratio >1:1, it yields P63+ LPC purity exceeding 80%. Moreover, the device consistently recovers >80% of P63+ LPCs, with the unrecovered fraction enriched in senescent cells exhibiting compromised clonogenicity and differentiation capacity. In direct benchmarking against DED and FACS, the MDDS device delivered a balanced performance in terms of purity and recovery, while offering advantages in throughput, consistency, and scalability. We propose that this technology could enable more consistent and efficient enrichment of feeder-cultured P63+ LPCs, thereby supporting more robust clinical manufacturing processes.
Sulfide solid electrolytes (SSEs) hold great promise for all-solid-state batteries (ASSBs), owing to their high ionic conductivity and excellent deformability. However, their practical application is severely hindered by high cost, primarily originating from lithium sulfide (Li2S), which accounts for ∼90% of the total SSE cost. Here, we report a novel strategy to produce low-cost Li2S from lithium carbonate (Li2CO3) via its reaction with ammonium thiocyanate (NH4SCN). This reaction generates only gaseous by-products, eliminating purification procedures and enabling scalable production of high-quality Li2S. The resulting Li2S enables the synthesis of representative SSEs, Li5.4PS4.6Cl0.8Br0.8 (LPSCB) and Li5.4PS4.6Cl1.6 (LPSC), with room-temperature ionic conductivities of 11.33 and 7.94 mS cm-1, respectively. When coupled with LiNbO3-coated LiNi0.895Co0.077Mn0.028O2 cathode, ASSBs deliver discharge capacities of 192.2 and 198.8 mAh g-1 at 0.1C, and retain 94.85% and 94.89% of initial capacities after 800 cycles at 1C, respectively. Cost analysis reveal that the total cost of SSEs synthesized from this Li2CO3-derived Li2S is reduced by 86.6% and 88.5%, highlighting its significant techno-economic advantages for commercializing SSEs toward ASSBs.
Structurally colored materials that change color upon mechanical stimuli have broad potential in stress sensors, wearable devices, and artificial skin. Such materials are conventionally prepared through the self-assembly of colloidal nanoparticles or block copolymers, a process that often involves time-consuming synthesis and purification as well as color inhomogeneity. To enable a simple and scalable route to mechanochromic materials, we employ holographic photopolymerization to fabricate monolithic, structurally colored elastomers. High reflectance and elasticity are achieved by adjusting the matrix formulation, the concentrations of recording monomers and acrylate cross-linker, the film thickness, and the exposure dose. The resulting materials exhibit a relative reflectance above 90% and a reversible mechanochromic shift of approximately 70 nm under 20% tensile strain. This work offers a practical approach to developing mechanically responsive photonic materials for sensing and anticounterfeiting applications.
As a finite strategic resource, helium is extracted from natural gas (NG). The concentration of helium in NG is very low, which makes helium hard to separate. The hydrate-based gas separation (HBGS) was proposed as a promising method for the separation of the NG with low helium content in this work. This work systematically investigated the HBGS of helium from simulated NG. The thermodynamic analysis reveals that the existence of 5.00 mol% tetrahydrofuran (THF) in the liquid phase decreased the gas-liquid-hydrate equilibrium pressure by 92.11%, compared to the deionized water system. The single-stage HBGS experimental results show that high THF concentration, low temperature, and high pressure benefited the gas processing capacity and helium purification, but they led to a low helium recovery rate. The best HBGS performance was limited by the "hydrate shell effect". The decrease in gas-liquid ratio led to an increase in helium concentration without losing the gas processing capacity, but it caused a decrease in the helium recovery rate. Through three-stage HBGS optimization, the helium concentration was increased from 0.54 mol% to 13.54 mol% (a 25.07-fold enrichment), and a total helium recovery of 87.34% was achieved. The mathematical model proposed in this work accurately predicts the performance of HGBS with 2.09% average relative error compared to the experimental data.
The assessment of developability and physical stability of antibody therapeutics is a cornerstone of modern biopharmaceutical discovery, increasingly relying on high-throughput automation to screen candidates to identify the most suitable lead candidate. In this study, we report a severe, anomalous fragmentation event observed in therapeutic antibodies during routine automated buffer exchange into phosphate-buffered saline (PBS) at physiological pH. Through a systematic investigation, we demonstrate that this instability is not intrinsic to the proteins, but is driven by the specific physicochemical environment of the purification process. We identify that at pH 7.4, the dominant phosphate species may act as a ligand capable of mobilizing trace catalytic copper from disposable chromatography columns. Our data suggests the leached copper coordinates to the antibody hinge region, driving a highly localized, Fenton-like oxidative cleavage. Comparative analysis reveals that this vulnerability is influenced by molecular architecture. Complex formats, such as 2 + 1 CrossMabs, exhibited significantly higher susceptibility than standard IgGs, suggesting that steric crowding may enhance the accessibility of the metal-binding site. Finally, we present a robust mitigation strategy using an ethylenediaminetetraacetic acid-based column conditioning protocol that effectively eliminates the leachable catalyst. These findings highlight a critical, often-overlooked source of chemical instability in automated workflows and underscore the necessity of controlling process-related impurities, particularly when developing complex antibody modalities.
Multicomponent reactions (MCRs) play a significant role in organic synthesis, medicinal chemistry and in material science, as they permit the formation of a single product from the strategic combination of more than two starting materials. Multicomponent reactions (MCRs) represent an attractive area of research for the efficient synthesis of highly functionalized, biological active and structurally diverse heterocyclic organic compounds. Synthesis of naphthoxazine-3-one derivatives is an important multicomponent reaction. Such derivatives are typically prepared in a one-pot from β-naphthol, aromatic aldehydes and urea in the presence of suitable catalyst thereby providing most efficient route for the synthesis of structurally diverse naphthoxazinone derivatives. Aromatic condensed naphthoxazinone derivatives represent an important class of functionalized building blocks. Naphthoxazinone derivatives are well-known by their significant biological activities such as anti-inflammatory, anti-ulcer, antibacterial, antipyretic, antifungal, antihypertensive, etc. These compounds also exhibit a wide range of industrial, clinical and pharmacological applications. Several researcher groups have made notable contributions in the field of synthesis of naphthoxazine-3-one derivatives using various catalytic systems. This review highlights the advances made over the past two decades in catalytic approaches, including nanocatalysis, for the synthesis of biologically active naphthoxazine-3-one derivatives. Especially, it emphases on methodologies reported by various researchers involving multicomponent cyclocondensation reactions of β-naphthol, aromatic aldehydes, and urea under specific reaction conditions. Most of the reported protocols have significant advantages like use of non-toxic and inexpensive catalyst, shorter reaction time, solvent-free conditions, easy work-up and purification of compounds, good to excellent yields of the desired naphthoxazine-3-one derivatives, recyclability of the catalysts, making green and environmentally benign protocols. This review will be definitely beneficial for researchers engaged in designing more efficient, novel and environmentally friendly synthetic protocols for the synthesis of naphthoxazine-3-one derivatives as well as for developing new biologically active heterocyclic molecules.
The growing threat of antibiotic resistance underscores the urgent need to investigate alternative antimicrobial agents, including bacteriocins or bacteriocins like peptides. The purpose of the present work was to isolate and characterize bacteria from sheep milk that produce antibacterial substances. A strain identified as Mammaliicoccus sciuri 1SH was isolated and its extracellular peptides were extracted and partially purified by ethyl acetate. Sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS-PAGE) analysis revealed a substance with a molecular weight of about 70 kDa. Activity was characterized across varying pH (2.5-8.5) and temperature (25-80 °C) ranges, demonstrating optimal activity between 25-37 °C and pH 6.5-7.4. Enzyme treatment with pepsin and pancreatin significantly reduced the activity, confirming its proteinaceous nature. The antibacterial substance demonstrated antibacterial activity against 26 clinical isolates of Escherichia coli, with minimum inhibitory concentration (MIC) values ranging from 1.125-70 µg/mL and minimum bactericidal concentration (MBC) 6.5-75 µg/mL. Growth kinetic tests demonstrated that growth of Escherichia. coli was suppressed in a dose-dependent way. Mechanistic studies revealed that the peptide induces DNA and protein leakage, indicating membrane disruption as a potential mode of action. Furthermore, the peptide demonstrated significant anti-oxidant activity (IC50: 13.5 μg/mL) and anti-inflammatory properties (IC50: 11 μg/mL). Fourier Transform Infra-Red (FT-IR) analysis identified key functional groups associated with the peptide. This study marks the first characterization and partial purification of a peptide derived from M. sciuri isolated from sheep milk, underscoring its potential as a promising antimicrobial agent with additional functional attributes.
In this study, we developed an optimized process targeting the key challenges in extracting astaxanthin from fresh H. pluvialis biomass. First, we compared the effectiveness of several commonly used cell disruption methods (acidic and alkaline hydrolysis, ultrasound, enzymatic treatment, and freeze-thaw cycles) on astaxanthin yield and quality. We then evaluated the effects of polar (alcohols: methanol, ethanol, and glycerol) and non-polar solvents (argan, cannabis, and olive oils) on astaxanthin extraction under different conditions (temperature and extraction time). Alkaline treatment with 1% KOH at 65 °C for 15 min provided the highest astaxanthin yield (65 mg/g), and astaxanthin recovery in methanol at 25 ± 1 °C reached 35.36 mg/g after 3 h. A biphasic methanol/olive oil solvent (0.6:1 v/v) enabled recovery of astaxanthin-enriched olive oil with a remarkable concentration of 125 mg/g in the oil phase. This protocol enables efficient astaxanthin extraction from wet biomass, eliminating the need for drying, which often leads to pigment loss, and is readily scalable for industrial applications. The optimized process represents a promising approach for astaxanthin recovery, with potential applications in the cosmetics industry.
Early and accurate detection of Pseudomonas aeruginosa (P. aeruginosa) is critically important in perioperative care to prevent severe healthcare-associated infections and guide timely antimicrobial intervention. Herein, we report a novel biosensing strategy for sensitive and label-free detection of P. aeruginosa by integrating F23 aptamer-mediated target recognition, garland rolling circle amplification (RCA)-triggered self-priming extension, and SYBR Green I (SG-I)-based fluorescence readout. The capture probe, comprising the F23 aptamer and a primer strand immobilized on magnetic nanoparticles, specifically recognizes P. aeruginosa and releases the primer to initiate dumbbell probe circularization and subsequent RCA. The resulting RCA products are cleaved by a nicking endonuclease to generate fragmented DNA, which then hybridizes with a hairpin probe to prime cyclic self-extension reactions, producing abundant double-stranded DNA for SG-I intercalation and fluorescence enhancement. Under optimized conditions, the proposed method achieves a detection limit as low as 2.3 CFU/mL with a wide linear range from 10 to 106 CFU/mL. The assay exhibits excellent specificity against non-target bacteria, robust stability during storage, and satisfactory anti-interference capability in complex clinical matrices. Validation using clinical samples demonstrates excellent agreement with the gold-standard colony counting method while reducing the assay time to less than 2.5 h without requiring nucleic acid extraction or thermal cycling. With its label-free design, isothermal amplification, and operational simplicity, this strategy holds great promise for point-of-care testing in perioperative settings and can be readily adapted for detecting other pathogens by substituting the corresponding aptamer.
Unmapped sequencing reads in livestock often contain valuable information about pathogens but are typically discarded. We analyzed blood-derived DNA and RNA from chickens and pigs kept under high- and low-biosecurity conditions, focusing on unmapped reads. In chickens, low-biosecurity farms harbored substantially more viral sequences, primarily plant viruses, indicating environmental contamination. In pigs, Mycoplasmoides pneumoniae and several pig-specific viruses were detected. Our bioinformatics pipeline, involving host read removal, assembly, BLAST, and taxonomic filtering, efficiently identified candidate pathogens and contaminants. This approach demonstrates the potential of sequencing-based environmental DNA monitoring to track microbial and viral presence, assess farm biosecurity, and support animal health surveillance.
Baichuan Baile (BCBL) is a novel herbal formula composed of three herbs and a food additive with proven antidepressant-like effects. However, its unclear bioactive constituents hinder quality control. This study evaluated BCBL-CP, a fraction extracted via ethanol precipitation and macroporous resin chromatography, mainly comprising coumarins (5-hydroxyxanthotoxin, oxypeucedanin hydrate, byakangelicin) and phthalides (senkyunolides H and I). We rapidly assessed BCBL-CP's efficacy using behavioral despair and 5-hydroxytryptophan-induced head-twitch tests. Subsequently, therapeutic effects on a reserpine-induced depression-like mouse model were evaluated through behavioral assessments. Its effects on the monoaminergic system, cerebral pathology, brain-derived neurotrophic factor (BDNF), and gut microbiota were detected through enzyme-linked immunosorbent assay, hematoxylin and eosin staining, immunofluorescence, and 16S rDNA sequencing. Results showed BCBL-CP exhibited significant antidepressant-like activity in both acute behavioral screening and chronic reserpine-induced depression studies. Mechanistically, BCBL-CP selectively increased prefrontal norepinephrine (p < 0.05), attenuated neuronal damage (p < 0.001), and showed a trend toward BDNF upregulation (p = 0.083). Uniquely, it restored gut microbiota α-diversity (p < 0.01), normalized β-diversity, and modulated eight key bacterial taxa (p < 0.01 or p < 0.05). These findings confirm that these five compounds constitute the major characteristic bioactive constituents of BCBL, providing quality-control markers for developing this prospective antidepressant.
An extreme disturbance, i.e. a 1-day chloroform fumigation, had a strong impact on the bacterial community structure in an extreme saline alkaline soil, but how Archaea respond to such an event is largely unknown. Three alkaline saline soils with electrolytic conductivity (EC) between 139 and 157 dS m- 1 and pH 10.0-10.3 were chloroform fumigated for one day and the recovery of the archaeal community determined after 1, 5 and 10 days, while an unfumigated soil served as control. Six archaeal phyla dominated by Candidatus Halobacterota (relative abundance 96.9%) were detected, while Natronorubrum dominated in the unfumigated (9.7%) and fumigated extreme alkaline saline soil (8.9%). Fumigation and time had a significant effect on the archaeal community structure. Some archaeal groups, e.g. Halovarius sp., were strongly affected by fumigation most accentuated on day 1 and 5. It was found that chloroform fumigation had only a limited and mostly a short time effect on the archaeal groups which demonstrated their resistance to a severe disturbance in extreme adverse conditions.
Despite significant advancements in medical and technical aspects in the field of dialytic renal replacement therapies, morbidity and mortality are extremely high among patients with end-stage renal disease (ESRD), and most interventional studies yielded unsatisfactory outcomes. Hemodiafiltration with endogenous reinfusion (HFR) emerges as a distinctive blood purification technique characterized by a dual-chamber dialyzer and a resin adsorption filter core, integrating the mechanisms of diffusion, convection, and adsorption within a single therapeutic framework. Current evidence suggests that, in addition to its capacity for the removal of uremic toxins, HFR could also be effective in ameliorating the inflammatory states and oxidative stress among the dialysis population; however, the existing evidence is limited by small sample sizes and short follow-up periods. Nevertheless, in clinical practice, several challenges associated with HFR in clinical practice necessitate immediate consideration. These challenges include the technical specifications of dialyzers, the impact on micronutrient levels, and the requirement for a broader scope of clinical indications. The present review elucidates the technical aspects of HFR and summarizes our perspectives on the contentious issues pertaining to its clinical application. Finally, we offer insights into future clinical trials exploring more specific indications for the utilization of HFR. Future studies should be adequately powered, prospective, multi-center, randomized controlled trials with robust clinical endpoints to address the identified challenges associated with HFR, thereby optimizing its therapeutic role in the management of ESRD. What is the impact of hemodiafiltration with endogenous reinfusion on nutrients retention?Are hemodiafiltration with endogenous reinfusion provide greater clearance efficiency on uremic toxins compared with hemoperfusion?Should hemodiafiltration with endogenous reinfusion be prescribed in patients with specific needs and more indications?