Near-infrared (NIR) transparent optical filters show great promise in night vision and receiving windows. However, NIR optical filters are generally prepared by laborious, environmentally unfriendly processes that involve metal oxides or petroleum-based polymers. We propose a lignin capturing–fusing approach to manufacturing optical biofilters based on molecular collaboration between lignin and cellulose from waste agricultural biomass. In this process, lignin is captured via self-assembly in a cellulose network; then, the lignin is fused to fill gaps and hold the cellulose fibers tightly. The resulting optical biofilter featured a dense structure and smooth surface with NIR transmittance of ~90%, ultralow haze of close to 0%, strong ultraviolet-visible light blocking (~100% at 400 nm and 57.58% to 98.59% at 550 nm). Further, the optical biofilter has comprehensive stability, including water stability, solvent stability, thermal stability, and environmental stability. Because of its unique properties, the optical biofilter demonstrates potential applications in the NIR region, such as an NIR-transmitting window, NIR night vision, and privacy protection. These applications represent a promising route to produce NIR transparent optical filters starting from lignocellulose biomass waste. Near-infrared (NIR) transparent optical filters with low haze and dark color have attracted interest in applications such as night vision, privacy protection, and medicolegal identification ,2. TheseLignocellulose-based biosourced materials are abundant, renewable, nontoxic, and mechanically strong candidates for optical materials –13. HoweTo address these essential needs, we propose a facile lignin capturing–fusing strategy to construct an NIR transparent lignocellulose-based optical biofilter directly from waste corncob Fig. A and B. In our lignin capturing–fusing approach, a key requirement is enabling a viscous flow state of the lignin in the cellulose network. Therefore, we used acetic acid pulping to process waste corncob into cellulose and partially acetylated lignin Notes  and S2. Tg) and lower than the viscous flow temperature (Tf) of lignin . FTIR spectroscopy  decisively confirmed the transformation of lignin and fused lignin and the interaction between fused lignin and cellulose , acetylated Aα (δC/δH of 72.6/4.86), and acetylated Aβ (δC/δH of 80.1/3.95) disappeared in the fused lignin, which demonstrated the cleavage of the acetyl groups and β−O−4 ether bonds. The lignin acetyl groups were detached because of the harsh pressure and temperature. This detachment led to acidolysis of lignin that eventually produced ketones and phenols, resulting in newly available hydroxyl groups that strengthened hydrogen bonding between the lignin and cellulose in the optical biofilter  spectroscopy and opy Fig.  showed td lignin . In addid lignin . The NMRose Fig. K to M. ASmooth surface and dense interior are key factors to realize the transparency of the optical biofilter. A smooth surface enabled the biofilter to reduce light reversal because of a uniform surface height that was attributed to the surface rearrangement during lignin fusion Fig. A and B. To further evaluate the unique optical properties of the biofilter, we measured the transmittance 200 to 1,400 nm) and haze (550 nm) of the films. UV-vis-NIR spectrophotometry demonstrated that lignin capturing process was the major contributor to the UV-blocking property of both lignin–cellulose film and optical biofilter, enabling resistance of almost all UV light , dioxane, acetone, ethyl ether, nol Fig. E. The binol Fig. . Comparenol Fig. F. We alsnol Fig. , which dnol Fig.  and S32.nol Fig. . ThroughGiven the excellent properties that met the demands of practical NIR transparent materials, we tested applications of the optical biofilter. To prevent blue light damage to the eyes, we fashioned the biofilter into eyeglass lenses Fig. . In addiWe demonstrated a novel lignin capturing–fusing approach to manufacturing an optical biofilter for use as NIR transparent material. The captured lignin was fused to fill the gaps in a cellulose network, which then held the fibers tightly and created a homogeneous dense structure. Both the lignin and the dense structure provided the biofilter with unique optical properties, including strong UV-vis light blocking (~100% at 400 nm and 57.58% to 98.59% at 550 nm), low haze (close to 0%), and high NIR transmittance (~90%). In addition, the biofilter had excellent, comprehensive stability, including water stability, solvent stability, thermal stability, and environmental stability. Thus, the multifunctional optical biofilter had many applications in optics, especially in the NIR region, such as reducing blue light, forming an NIR transmitting window for fruit detection, preventing an NIR night vision monitor from glare blindness, and protecting sensitive information. This proposed lignin capturing–fusing approach for high-performance optical biofilter fabrication demonstrates a pathbreaking opportunity for lignocellulose-based biomass to broaden applications in optics.3NOH; 40 wt% of aqueous solution) was purchased from TCI Shanghai Co. Ltd. . Ethylene glycol (98 wt%), tert-butanol (≥99.5%), and anhydrous calcium chloride  were supplied by Shanghai Macklin Biochemical Co. Ltd. . Glacial acetic acid, sodium acetate, sulfuric acid, hydrochloric acid, dioxane, acetone, ethyl ether, N,N-dimethylformamide, ethanol, sodium hydroxide, and urea were purchased from Sinopharm Chemical Reagent Co. Ltd. . Cellulose and lignin were extracted from corncob waste (Notes Benzyl-trimethyl ammonium hydroxide (BzMete Notes  and S2.3NOH (22.5 g) at 8 °C. The mixture was further incubated at 8 °C for 24 h, centrifuged at 6,000 rpm for 5 min, cast with a height of 1 mm, and regenerated in ethylene glycol at 8 °C. The resulting cellulose hydrogel was washed with deionized water to remove chemicals. Then, the hydrogel was immersed in aqueous 5 wt% of lignin/acetic acid (90 vol%) for 5 h (hydrogel: lignin solution ≈ 1 g: 20 ml) to exchange the solution, followed by immersion in deionized water to remove acetic acid (lignin capturing process). The subsequent lignin–cellulose hydrogel was dried at 97 °C using a rapid Köthen sheet former  to manufacture a lignin–cellulose film. Finally, the lignin–cellulose film was hot-pressed between 2 polyimide films at 180 °C and 5 MPa for 1 h to produce optical biofilter (lignin fusing process). Biofilters with different lignin loading contents were produced by using acetic acid solutions with lignin concentrations of 1, 5, and 10 wt% and named optical biofilter_1%, optical biofilter_5%, and optical biofilter_10%  extracted from corncob was swelled in deionized water (7.5 g) and dissolved with 40 wt% of aqueous BzMe10% Note . All tes10% Note  to S7.−1 and 2-dimensional heteronuclear single quantum coherence (2D-HSQC) NMR . Fused lignin was hot-pressed for 1 h  for the 2D-HSQC NMR measurement. X-ray diffraction  with a Cu Kα monochromatic radiation source was used to characterize crystallization conditions with the 2θ angle from 5° to 40° and a scanning speed of 5°·min−1. Differential scanning calorimetry  was performed with a heating rate of 10 °C·min−1 in a nitrogen atmosphere; all samples were first heated at 105 °C for 10 min before testing from 30 to 200 °C. NIR diffuse reflectance spectra in the range of 4,000 to 10,000 cm−1 were obtained with an Fourier transform near-infrared analyzer .The surface and cross-profile morphology of samples were observed by scanning electron microscopy  (5 kV) and confocal Raman spectroscopy . Molecular structures were determined by FTIR  in the range of 650 to 4,000 cmTransmittance and haze were measured by UV-Vis-NIR  with a wavelength range of 200 to 1,400 nm and a transmittance/haze tester  with a wavelength of 550 nm.2 in a sealed bottle (120 h) in air atmosphere of saturated salt water (25 °C). The weight change of CaCl2 was used to calculate water vapor transmittance rate, equivalent to the ratio of the mass change of CaCl2 to the effective area for 1 d (in grams per square meter per day). Water absorption was measured by the weight before and after immersing cellulose film, lignin–cellulose film, and optical biofilter in deionized water for 30 min. Then, the shape stability was illustrated by recording the state of drying, wetting, and redrying. Water contact angle was measured using a contact angle measuring system  in 180 s at ambient temperature. Solvent stability was evaluated by immersing the optical biofilter for 21 d in deionized water, 90 vol% of aqueous acetic acid, dioxane, acetone, ethyl ether, N,N-dimethylformamide, ethanol, 1 wt% of aqueous NaOH, and tert-butanol. Thermal stability was measured by a thermogravimetric analyzer  from 100 to 700 °C with a heating rate of 20 °C·min−1 and a nitrogen atmosphere. Environmental stability of the optical biofilter with size of 20 mm × 20 mm was investigated in artificial soil environment and simulated environmental changes with deionized water and sun exposure for 150 d.Water vapor barrier properties were measured by the color change of silica gel in a sealed bottle (48 h) and calculated by the weight change of CaCl−1.The tensile properties of cellulose film, lignin–cellulose film, and optical biofilter, 30 mm × 10 mm (length × width), were measured using an electromechanical universal testing machine  equipped with a 250-N load cell and a constant speed of 5 mm·min


A previous trial of dapagliflozin led to no improvement in weight or glycemic control. Semaglutide was introduced (0.25 mg weekly increased to 0.5 mg weekly) and was well tolerated. After 6 months, weight had decreased by 16 kg and insulin dose by 36%. Despite less insulin, hemoglobin A1c improved, with reduced glycemic variability and no increase in hypoglycemia. Semaglutide may exert significant metabolic benefits in patients with established T1DM, even where c-peptide is no longer detectable. This case supports the need for a dedicated trial examining potential benefits of semaglutide in T1DM.The efficacy of glucagon-like peptide-1 receptor agonists in type 2 diabetes is well established, but their role in type 1 diabetes (T1DM) is less clear. A 36-year-old woman with a 27-year history of T1DM and undetectable c-peptide presented for review of weight management, with body mass index 29.3 kg/m The efficacy and safety of the glucagon-like peptide-1 (GLP-1) receptor agonist semaglutide in type 2 diabetes (T2DM) is well established. In patients with T2DM, semaglutide leads to weight loss, improvement in systolic blood pressure, and a reduction in composite cardiovascular outcomes , 4. The A 36-year-old woman presented for review of T1DM. The patient described extreme difficulty in losing weight and persistent glycemic variability. She had been diagnosed 27 years earlier (aged 9 years). There were no known macrovascular complications; microvascular complications included stable, inactive proliferative diabetic retinopathy previously treated with bevacizumab and laser photocoagulation. Diabetes management included multiple daily injections of insulin, insulin glargine 300 units/mL 21 U daily and insulin lispro 16 U daily. Additional background history was significant for polycystic ovary syndrome (PCOS) treated with metformin 1000 mg twice daily and an oral contraceptive pill. Glucose monitoring was performed using the Freestyle Libre flash glucose monitoring system.2. Blood pressure was 137/86 mmHg. Hemoglobin A1c (HbA1c) was 7.2% (55 mmol/mol) with fasting c-peptide <0.05 nmol/L . Urine albumin creatinine ratio was 3.3 mg/mmol . Creatinine was 70 µmol/L  with estimated glomerular filtration rate >90 mL/min/1.73 m2. Random total cholesterol was 4.8 mmol/L  with triglyceride 1.4 mmol/L .At the time of presentation, her weight was 78.8 kg and body mass index 29.3 kg/mWith informed consent, there had been previous trial of a sodium glucose cotransporter-2 inhibitor, dapagliflozin 10 mg per oral daily. However, after 3 months, dapagliflozin had been ceased as there was minimal effect on glucose variability, no change in insulin doses, and no appreciable weight loss.At the patient’s request, and with informed consent, treatment with semaglutide was initiated with a starting dose of 0.25 mg weekly subcutaneously. The dose was uptitrated to 0.5 mg weekly, which was associated with reduced appetite without significant nausea or gastrointestinal disturbance. Metformin 1000 mg twice daily was continued as management for PCOS. Retinal screening was performed before initiation of semaglutide to ensure retinopathy was inactive and stable compared with previous assessments.After 6 months of treatment, the patient’s weight had decreased to 63 kg (−16 kg) and insulin doses were reduced by 43% . Systolic blood pressure reduced by 18 mmHg. Despite less insulin, HbA1c improved to 6.3% (−0.9%) with no increase in hypoglycemia. Changes in clinical and biochemical parameters before and after treatment with semaglutide are summarized in After 6 months of use, because of supply chain issues and inability to source semaglutide, her treatment was interrupted for 4 weeks, during which there was a deterioration in glycemia and increased glucose variability .Insulin remains the mainstay of treatment for T1DM. However, adjunctive treatments such as the GLP-1 receptor agonists may provide additional therapeutic options through direct and indirect effects on glucagon release and insulin resistance.T1DM is characterized by pancreatic α-cell dysfunction in addition to β-cell dysfunction . AccordiInsulin resistance can also occur in T1DM by exogenous subcutaneous insulin injection bypassing first-pass hepatic insulin clearance, possibly by insulin receptor downregulation and/or promotion of abdominal adiposity through stimulation of lipogenesis . In addiThere is conflicting evidence regarding residual pancreatic function on the effect of GLP-1 in T1DM. When ultrasensitive assays are used, most patients with more than 5 years of T1DM have detectable fasting c-peptide . FurtherPatients with T1DM should be counselled about the potential adverse effects of GLP-1 receptor agonists. Gastrointestinal side effects of GLP-1 receptor agonists are well recognized, with dose escalation used to minimize the risk. Our patient tolerated semaglutide, in addition to metformin, without gastrointestinal side effects, which suggests that, as in patients with T2DM, patients with T1DM likely will experience a spectrum of gastrointestinal effects related to GLP-1 receptor agonists. Another consideration is the effect of GLP-1 receptor agonists, particularly semaglutide, on diabetic retinopathy. In T2DM, semaglutide 1.0 mg weekly was associated with deterioration in diabetic retinopathy, although this is deemed to be secondary to rapid glucose improvements rather than a direct effect of GLP-1 receptor agonists per se [Our patient was overweight and had comorbid PCOS before initiation of semaglutide, a condition associated with increased body mass index and insulin resistance. GLP-1 receptor agonists, either as monotherapy or in combination with metformin, are associated with weight loss and improved metabolic markers in women with PCOS and overweight or obesity . This isA higher dose of semaglutide, 2.4 mg weekly, has been associated with weight loss in people with obesity without diabetes [Currently, there is only 1 report of semaglutide use in a patient with T1DM that demonstrated reduction in weight, insulin requirement, and HbA1c similar to reductions seen in our patient; that study did not report the c-peptide status of the patient . Our casAuthor Contributions: L.R.: involved in manuscript writing and submission. C.M.: involved in the management of this patient and manuscript writing. J.G.: involved in manuscript writing. All authors reviewed and approved the final draft.Insulin is the mainstay of treatment for type 1 diabetes; however, adjunctive therapies should be considered and further studiedThere may be metabolic benefits to the addition of glucagon-like peptide-1 receptor agonists in patients with type 1 diabetesThe benefits of glucagon-like peptide-1 receptor agonists may not be limited to patients with detectable c-peptide levelsThe safety of glucose-like peptide 1 receptor agonists needs to be assessed further


HIV continues to affect millions of men and women worldwide. The development of long-acting injectables for HIV prevention can overcome adherence challenges with daily oral prevention regimens by reducing dosing frequency and stigma. We previously developed an ultra-long-acting injectable, biodegradable, and removeable in situ forming implant (ISFI) with cabotegravir (CAB) that demonstrated protection after multiple rectal SHIV challenges in female macaques. Here, we sought to further characterize CAB ISFI pharmacokinetics (PK) in mice by assessing the effect of dose and number of injections on CAB PK, time to completion of CAB release and polymer degradation, long-term genital tissue PK, and CAB PK tail after implant removal. CAB concentrations in plasma were above the benchmark for protection for 11–12 months with proportionality between dose and drug exposure. CAB ISFI exhibited high concentrations in vaginal, cervical, and rectal tissues for up to 180 days. Furthermore, depots were easily retrievable up to 180 days post-administration with up to 34% residual CAB and near complete (85%) polymer degradation quantified in depots ex vivo. After depot removal, results demonstrated a median 11-fold decline in CAB plasma concentrations across all doses. Ultimately, this study provided critical PK information for the CAB ISFI formulation that could aid in its future translation to clinical studies. Globally, there are over 38 million people living with HIV with approximately 1.5 million new infections reported in 2021 ,5. In December 2021, the FDA approved the first long-acting injectable containing integrase inhibitor cabotegravir (CAB LA) for HIV PrEP given as bimonthly intramuscular injections . Although CAB LA represents a major progression in the HIV prevention landscape, there are a few limitations with the technology, such as large injection volumes (3 mL), injection site reactions ,7 and neco-glycolic acid) (PLGA)), water-miscible organic solvents  or dimethyl sulfoxide (DMSO)), and active pharmaceutical ingredients (APIs). When co-formulated, these components generate a syringeable and homogenous liquid suspension for injection into the subcutaneous or intramuscular space. Upon injection, a phase inversion occurs resulting in a solid depot with the API dispersed within the precipitated polymer matrix [50) and the no-observable-effect-level (NOEL) when administered to macaques [In situ forming implants (ISFIs) may overcome limitations associated with CAB LA due to their ability to achieve ultra-long-acting drug release, compatibility with many drugs including ARVs, biodegradable nature, and ability to be safely removed via a small incision to terminate treatment, if necessary ,12,13. Ir matrix ,15,16. Dr matrix . In our r matrix ,13,17,18r matrix , which cmacaques ,22,23,24Specifically, we previously developed a subcutaneously administered ultra-long-acting (>6 months) and well-tolerated PLGA-based ISFI with CAB (500 mg/mL) that demonstrated complete protection in female rhesus macaques after multiple rectal simian–human immunodeficiency virus (SHIV) challenges . FurtherThus, we sought to perform a comprehensive dose-ranging PK study in mice with CAB ISFI (500 mg/mL) to investigate (1) CAB ISFI time to completion, (2) long-term (180 days) vaginal and rectal tissue PK, (3) CAB PK tail post-depot removal, (4) implant biodegradation, and (5) the effect of dose (50 µL (1215 mg/kg) vs. 100 µL (2430 mg/kg)) and number of injections (100 µL vs. 2× 50 µL) on CAB in vivo drug release . This stPoly containing 50:50 lactic acid:glycolic acid was purchased from LACTEL . N-methyl-2-pyrrolidone  was received from ASHLAND . Dimethyl sulfoxide  was purchased from Sigma-Aldrich . HPLC grade acetonitrile (ACN) and water were purchased from Sigma Aldrich . Tetrahydrofuran (THF) was purchased from Sigma Aldrich . High purity (≥99%) CAB was purchased from Selleckchem .A reverse-phase HPLC analysis was performed on an Agilent 1260 HPLC system  equipped with a Diode Array Detector and an LC pump with autosampler, as previously described . In briew/w) NMP:DMSO) at a ratio of 1:4 w/w. Solvents and placebo ISFI formulations were sterile filtered  prior to incorporating the drug in the formulation. Cabotegravir (500 mg/mL) was added to the sterile placebo solution producing a stable ISFI suspension and mixed at 37 °C for 48 h. To ensure homogeneity of the drug suspension, sample aliquots were collected from the drug formulation and dissolved in ACN. Drug concentration was quantified by HPLC analysis.The preparation of CAB ISFIs has been previously described . In brieAll in vivo studies were conducted with female BALB/c mice . All experiments involving mice were carried out with an approved protocol by the University of North Carolina Animal Care and Use Committee. Housing conditions consisted of a 12 h/12 h light/dark cycle. The ambient temperature was 68–72 degrees Fahrenheit with 30–70% humidity. All injections were administered subcutaneously with a 19 G needle. All samples were stored at −80 °C until PK analysis. A 367-day in vivo study was conducted to assess long-term and time-to-completion PK of CAB ISFIs in female BALC/c mice. Mice were subcutaneously injected with 50 µL, 100 µL, or 2× 50 µL of CAB ISFI formulation (n = 6/dose). For this study, 2.5 mL of CAB ISFI formulation was prepared to ensure sufficient volume during injections, and an analytical CAB concentration of 499.1 ± 12.7 mg/mL was determined via HPLC analysis. At 1 h, 1 day, 3 days, 7 days, 30 days, and monthly thereafter, plasma samples were collected longitudinally from mice. It is important to note that CAB plasma concentrations up to 90 days post-injection for mice injected with 50 µL of CAB ISFI have been previously reported . The datArea-under-the-curve (AUC) for all injection volumes were calculated using non-compartmental analysis (NCA) in Phoenix WinNonlin (version 8.3) . The log-linear trapezoidal method was used to calculate AUCs over the 367-day time course. PK data are represented as median with interquartile range. ® hard tissue reinforced metal bead kit tubes  containing 1 mL of 70:30 acetonitrile:water, homogenized, and then centrifuged. Following protein precipitation extraction with the isotopically labeled internal standard, 13C,2H2,15N-CAB, CAB was separated using reverse-phase chromatography via a Waters Atlantis T3  analytical column. An AB Sciex API-5000 triple quadruple mass spectrometer was used to detect the analyte and internal standard under positive ion electrospray conditions. Precision and accuracy of the calibration standards and quality control samples were within 15% for the dynamic range of 1.00–4000 ng/mL. Final concentrations were normalized to tissue mass and reported in ng/g.A 180-day study was conducted to assess the distribution of CAB in vaginal, cervical, and rectal tissue in female BALC/c mice. Mice were subcutaneously injected with 50 µL, 100 µL, or 2× 50 µL of CAB ISFI formulation (n = 3/dose per timepoint). For this study, 6.25 mL of CAB ISFI formulation was prepared to ensure sufficient volume during injections, and an analytical CAB concentration of 499.3 ± 8.6 mg/mL was determined via HPLC analysis. At 7, 30, 60, 90, and 180 days post-injection, vaginal, cervical, and rectal tissues were collected from each mouse, weighed, and stored at −80 °C until PK analysis. Weighed tissues were transferred into PrecellysA 180-day in vivo study was conducted to assess plasma concentrations after CAB ISFI removal, residual CAB, and biodegradation of PLGA after ISFI removal in female BALB/c mice. Mice were subcutaneously injected with 50 µL, 100 µL, or 2× 50 µL of CAB ISFI formulation . For this study, 9 mL of CAB ISFI formulation was prepared to ensure sufficient volume during injections and an analytical CAB concentration of 497.2 ± 12.6 mg/mL was determined via HPLC analysis. At 1 h, 1 day, 3 days, 7 days, 30 days, 60 days, 90 days, and 180 days (n = 6 mice/timepoint per dose), plasma samples were collected longitudinally from mice. At days 30, 60, 90, and 180 (n = 6 mice/timepoint per dose), CAB ISFI was carefully removed via a small skin incision at the injection site, flash frozen, and stored at −;80 °C until further processing . Plasma CAB ISFIs were extracted to quantify the amount of CAB remaining in the implant after removal from mice at days 30, 60, 90, and 180 post-administration (n = 3/timepoint per dose). Each implant was massed to assess mass loss over time. To quantify residual drug concentration, implants were dissolved in ACN to extract residual CAB. Residual CAB from the implant was quantified by HPLC analysis. It is important to note that CAB residual drug measurements up to 90 days post-injection for mice injected with 50 µL of CAB ISFI have been previously reported . The datGel permeation chromatography (GPC) was conducted as previously described . In briep value of <0.05 (95% confidence level) was deemed significant.Statistical analyses were performed in GraphPad Prism version 9.4 . A Kruskal–Wallis one-way ANOVA followed by a Dunn’s multiple comparisons test was performed to analyze differences in drug exposure (AUC) in plasma across doses and number of injections. To assess differences in CAB concentration in vaginal, cervical, and rectal tissues across doses and number of injections, a two-way ANOVA and Tukey’s multiple comparisons test were conducted with respect to dose and timepoint. A 90, 664 ng/mL) for 11 months when injected with 50 µL of CAB ISFI and 12 months when injected with 100 µL or 2× 50 µL of CAB ISFI. Due to the longer than expected duration of the CAB ISFI, time to completion was not determined and this study was terminated at 367 days. However, mice that were injected with 50 µL of CAB ISFI elicited a 19-fold decline in CAB plasma concentrations between day 330 and 367, exhibiting average concentrations near the 1× PA-IC90 (166 ng/mL). We could expect the 100 µL and 2× 50 µL of CAB ISFI groups to follow a similar trend if the study continued for an additional couple of months. Dose-ranging ultra-long-acting PK studies in female BALB/c mice were conducted over 367 days to assess the duration of complete CAB release across different injection volumes and doses (50 µL (1215 mg/kg), 100 µL (2430 mg/kg), and 2× 50 µL (2430 mg/kg)). Mice received a subcutaneous injection of 50 µL, 100 µL, or 2× 50 µL (n = 6 mice/dose) of CAB ISFI (500 mg/mL). CAB plasma concentrations for each dose were analyzed and plotted over 367 days . Mice elp = 0.3372) (90) for 11–12 months reducing dosing frequency compared with marketed products and (2) CAB ISFI dose is approximately proportional to overall drug exposure and there is no difference in drug exposure when administering the same dose with multiple injections. Knowing these characteristics of the formulation, if CAB ISFI were to fall below the protective benchmark in large animal or human studies, an increase in injection volume could provide the ability to reach protective plasma levels due to proportionality with drug exposure. Furthermore, the results demonstrated proportionality between dose (1215 mg/kg (50 µL) vs. 2430 mg/kg (100 µL)) and overall CAB exposure (AUC) . Additio 0.3372) . Ultimatp > 0.05). CAB concentrations were measured in vaginal, cervical, and rectal tissues in female BALB/c mice up to 180 days post-CAB ISFI injection. These genital tissues represent main entry points for HIV infection; thus, it is crucial to ensure CAB concentrations are high in these target tissues to provide protection against HIV infection. Currently available CAB LA for HIV prevention demonstrated a PK tail for at least 15 months due to CAB’s long half-life (40 days) and inability to be removed to terminate treatment . InvestiResults showed that plasma CAB levels exhibited a significant decrease after implant removal across doses A–D. MeanWhen CAB ISFIs were removed 30 and 60 days post-administration, mice exhibited a rapid decline in CAB plasma concentrations within 7 days post-removal, but CAB concentrations reached a plateau between day 7 and day 30 post-removal A,B. InteTo assess CAB ISFI retrievability over time, we removed CAB ISFIs from mice via a small skin incision at the injection site at days 30, 60, 90, and 180 post-administration for each dose. Six mice per timepoint per dose were evaluated. However, 1 mouse that received a 100 µL injection for day 60 depot removal and 1 mouse that received a 2× 50 µL injection for day 180 depot removal were removed from the study prior to implant retrieval due to health concerns. ISFIs across doses were successfully removed from mice with no fibrotic tissue surrounding the depot A. For 1 Depots were further processed to evaluate residual CAB and polymer degradation. After 180 days in mice, there was up to 34% CAB remaining  and apprSince only 23–34% of CAB remained after 180 days, it would be unlikely for CAB ISFI to produce protective concentrations for an additional 6 months. However, as previously seen in 90 for 11–12 months, owing to its ultra-long-acting capability. Additionally, proportionality between CAB ISFI dose and overall drug exposure was observed. Furthermore, we demonstrated high CAB concentrations in vaginal, cervical, and rectal tissues for 180 days with no significant differences between doses or number of injections. When assessing the CAB PK tail, a significant decline in CAB plasma concentrations was observed within 30 days after depot removal, but complete CAB elimination was not achieved, which may be due to the presence of a CAB reservoir in the subcutaneous tissue. After 180 days post-injection, CAB ISFIs were easily retrievable with up to 34% CAB remaining and an 15% polymer remaining. These results confirm CAB ISFI’s ultra-long-acting ability with high concentrations in plasma and target genital tissues, which are common sites of HIV infection. Ultimately, this study further characterized the PK of the CAB ISFI formulation and can assist with determining optimal administration parameters for clinical translation. In future, we plan to perform a complete biodistribution study in mice and similar studies should be performed in macaques to confirm results with clinically relevant injection volumes and doses. We performed a comprehensive dose-ranging PK study of CAB ISFIs in mice and assessed the effect of dose and number of injections on CAB ISFI PK, time to completion of CAB release and polymer degradation, long-term genital tissue PK, and CAB PK tail post-depot removal. After a single administration of CAB ISFI, plasma concentrations were maintained above the 4× PA-ICSRB, ICY, and RS are inventors on a patent describing the Injectable Polymer-Based Drug Formulation for Ultra-Long-Acting Drug Delivery.


GBA1 variants are the strongest genetic risk factor for Parkinson’s disease (PD). However, the pathogenicity of GBA1 variants concerning PD is still not fully understood. Additionally, the frequency of GBA1 variants varies widely across populations.GBA1 variants in Norwegian PD patients and controls, and to review the current literature on newly identified variants that add to pathogenicity determination.To evaluate Oxford Nanopore sequencing as a strategy, to determine the frequency of GBA1 gene on the Oxford Nanopore GridION as an 8.9 kb amplicon. Six analysis pipelines were compared using two aligners  and three variant callers . Confirmation of GBA1 variants was performed by Sanger sequencing and the pathogenicity of variants was evaluated.We included 462 Norwegian PD patients and 367 healthy controls. We sequenced the full-length GBA1 variant calls, while 4.2% (5/120) variant calls were false-positive, with the NGMLR/Minimap2-BCFtools pipeline performing best. In total, 13 rare GBA1 variants were detected: two were predicted to be (likely) pathogenic and eleven were of uncertain significance. The odds of carrying one of the two common GBA1 variants, p.L483P or p.N409S, in PD patients were estimated to be 4.11 times the odds of carrying one of these variants in controls .We found 95.8% (115/120) true-positive GBA1 variants. Further studies on the pathogenicity of GBA1 variants are needed to assess their effect on PD.In conclusion, we have demonstrated that Oxford long-read Nanopore sequencing, along with the NGMLR/Minimap2-BCFtools pipeline is an effective tool to investigate The online version contains supplementary material available at 10.1186/s12864-023-09417-y. GBA1 (Glucocerebrosidase A) are considered a common genetic risk factor for PD [GBA1 classically cause Gaucher’s disease (GD) and an increased PD risk has been observed in patients with GD and asymptomatic carriers of heterozygous variants [GBA1 heterozygous variant compared to non-carriers, and it is even lower in GBA1 homozygotes/compound heterozygotes [GBA1 variants in PD include p.E365K , p.T408M , p.N409S , and p.L483P . However, the classification of the pathogenicity of GBA1 variants and their effect on PD is still ongoing. For this reason, the reported frequencies of GBA1 variants across studies are rather inconsistent, with frequencies ranging from 1.8% up to 47% depending on the ethnicity of the samples and the GBA1 variants investigated [GBA1 variants  [GBA1 variant were found: four of the patients had a p.N409S (1.3%) and three had a p.L483P (1.0%) substitution.Parkinson’s disease (PD) is a multifaceted and highly complex neurodegenerative disorder. Multiple genes have been implicated in PD. Variants in r for PD . Biallelvariants –10. Glucstigated . In a prGBA1 gene is the nearby pseudogene GBAP1. GBAP1 shares 96% exonic sequence homology with the GBA1 coding region with the highest homology between exons 8 and 11. In this region, most pathogenic variants have been reported, usually resulting from recombination events, e.g. gene conversion, fusion, or duplication [GBA1 is by long-read sequencing [GBA1 sequencing to detect exonic and intronic variants and recombinant alleles in combination with phase information, at high multiplex capacity [Another challenge that arises when sequencing the lication . This coquencing . This tecapacity , 16.GBA1 in a sample from the Norwegian population by (1) employing and evaluating Oxford Nanopore sequencing as a strategy, (2) determining the frequency of variants within the GBA1 gene in patients with PD and healthy controls by providing an update on previous reports [Herein, we have comprehensively characterized  reports , 17, andAfter Nanopore sequencing of the long-range PCR products, we obtained a mean read length of 5.2 kb SD = ± 2.1 kb) and a mean read quality Phred score of 14.2 (SD = ± 0.4) across all raw sequencing data. After length and quality filtering and read trimming, we obtained a mean read length of 9.0 kb (SD = ± 0.2 kb) and a mean read quality Phred score of 15.7 (SD = ± 0.5). For the filtered samples the mean coverage was 193.1X (SD = ± 187.5X), ranging between 20.6X and 1820.8X across all samples. Nevertheless, the coverage per sample was consistent over all positions  were detected in the Nanopore sequencing analysis after filtering. Out of the 79 rare GBA1 variants, 18 variants were categorized as “pathogenic”, “likely pathogenic”, or of “uncertain significance” . Of these, 15 rare variants, categorized as “pathogenic”, “likely pathogenic”, or of “uncertain significance”, were found in 111 samples , again independent of the aligner. After Sanger sequencing, 13 variants in 110 samples  were validated, indicating five false-positive calls but no false-negative calls.With BCFtools, 64 rare annotated GBA1 variants were detected in 308 samples, resulting in 426 calls. Of these calls, 117 calls referred to 14 rare variants, categorized as “pathogenic”, “likely pathogenic”, or of “uncertain significance”, and were detected in 108 samples. After Sanger sequencing, 112 calls in 107 samples were validated, implying that five calls were false-positive, with an additional three calls being false-negative. When Minimap2 was used as an aligner, again 65 rare annotated GBA1 variants were detected, however, here they were detected in 311 samples . With this pipeline, 15 rare annotated GBA1 variants, categorized as “pathogenic”, “likely pathogenic”, or of “uncertain significance”, were found in 110 samples . Of these, 113 calls in 108 samples could be validated with Sanger sequencing, indicating six false-positive calls and two more false-negative calls.For the pipeline using Clair3 as a variant caller after a preceding alignment with NGMLR, 65 rare annotated GBA1 variants were detected in 318 samples, including 123 calls of 17 rare variants, categorized as “pathogenic”, “likely pathogenic”, or of “uncertain significance”, in 114 samples. Only 108 calls in 103 samples were validated with Sanger sequencing, leading to 15 false-positive and additional seven false-negative calls. Similarly, when Minimap2 was used for the alignment, 76 rare annotated GBA1 variants were detected in 331 samples, resulting in 481 calls. Of these, 18 rare variants, categorized as “pathogenic”, “likely pathogenic”, or of “uncertain significance”, were detected in 120 samples . Here, 109 calls in 104 samples were Sanger validated, indicating 21 false-positive and six false-negative calls.Lastly, the Pepper-Margin-Deepvariant pipeline was used for variant calling. After preceding alignment with NGMLR, 453 calls of 72 rare annotated GBA1 were validated in 462 Norwegians with PD and 367 healthy controls  and eleven GBA1 variants were of “uncertain significance” . The total carrier frequency of rare GBA1 variants predicted as “pathogenic”, “likely pathogenic”, or of “uncertain significance” was 17.1% (79/462) in the PD cases and 8.4% (31/367) in the controls    . However, all samples tested negative for SVs.In addition, samples with a rare “pathogenic”, “likely pathogenic”, or “uncertain” GBA1 variants detected in PD and their frequencies across different populations. In total, 100 articles on GBA1 variant frequencies across populations were included in the overview  and in 6.6% of controls (29/419) , and p.T408M, found in 1.7% of patients with PD (7/442) and in 3.6% of controls (16/419) .We performed a systematic literature review to summarize GBA1 gene are known to affect PD risk, however, the frequency and pathogenicity of these variants are still under debate. The latter is further complicated as the frequencies of GBA1 variants vary by population. In our study, we used the pathogenicity scoring of ACMG, Varsome, ClinVar, SIFT, Polyphen2, CADD, and GERP + + and categorized the variants found into “pathogenic”, “likely pathogenic”, and variants of “uncertain significance”. Here we report 13 rare variants within GBA1 in Norwegian PD cases, with two of them predicted to be “pathogenic” or “likely pathogenic”, and eleven GBA1 variants of “uncertain significance”. In total, we found a “pathogenic” or “likely pathogenic” variant  in 1.5% of PD cases and in 0% of healthy controls. However, pathogenicity scoring with ACMG does not take the association with PD into account and consequently underestimates the frequency of risk variants in GBA1. Therefore, we further investigated known GBA1 risk variants associated with PD. In our study, 15.2% of patients with PD carried a common GBA1 risk variant , compared to 7.9% of controls.Variants in the GBA1 gene in patients with PD and in the general population in 100 studies , as well as three different variant callers . BCFtools performed best with regard to the number of true-positive, false-positive, and false-negative hits, independently from the aligner used. However, one limitation is that we could not fully evaluate pipeline sensitivity. As we only confirmed variants called by our Nanopore analysis pipelines by Sanger sequencing, we underestimate false-negative variants that were not initially called. With Oxford Nanopore technology we assessed the precision of long-read sequencing and consensus data analysis and detected 115 real GBA1 variant calls, while five variant calls were false-positives. Thus, > 95% of called variants were true-positive. Nanopore long-read sequencing is an accurate tool to detect genetic variations and with further development in flow cells and sequencing kits, the accuracy of variant detection is likely to increase. Another advantage of this technology is the capacity to multiplex samples, which decreases analysis costs of the full 8.9 kb GBA1 gene to $13 USD per sample, which is lower than other DNA sequencing methods [GBA1 gene without sequencing the pseudogene GBAP1, and to detect all disease-causing variants including information on phase [In addition to variable inclusion criteria, the sequencing method and data analysis, and the chronology when it was performed, has a major influence on the detection rate and  methods . A strenon phase .GBA1 variants. We thoroughly evaluated six different data analysis pipelines and found the pipeline consisting of an alignment with either NGMLR or Minimap2 and variant calling with BCFtools to perform best with regard to detecting variants in GBA1. With our established and validated workflow, we demonstrated that the frequency of the two common GBA1 risk variants for PD  in Norwegian patients with PD is 4.3% and higher than in the general population (1.1%). Furthermore, we reviewed current literature on GBA1 variant frequencies in PD across populations, thereby adding to pathogenicity determination. Given the importance of this gene, further functional studies on the pathogenicity of GBA1 are needed to assess their effect on PD.In conclusion, we have demonstrated that Oxford Nanopore sequencing is an efficient and scalable tool for investigating A sample of 462 Norwegian patients with PD was included in this study , described previously [We used blood-derived genomic DNA samples from all PD patients and controls. Informed consent was obtained from all participating individuals. We enriched for eviously  using thhttps://github.com/rrwick/Filtlong) to only include the best 50% of the reads, based on Phred quality scores (q-score) in the FASTQ files, with a minimum read length of 8 kb. Afterwards, the reads were trimmed with NanoFilt [GBA1 were annotated using ANNOVAR [Data acquisition and run monitoring was carried out with MinKNOW (version v21.05.25 and later). The integrated Guppy algorithm (version v5.0.16 and later) was used for base-calling with the super-accurate base-calling model, de-multiplexing, and FAST5 and FASTQ file generation. The base-called reads were filtered with Filtlong (v.0.2.0) (NanoFilt  (v2.8.0)NanoFilt  (v0.2.7)NanoFilt  (v2.22).NanoFilt  using v1NanoFilt  (v.1.15)NanoFilt  (v1.9), NanoFilt   due to the high exonic sequence homology between GBA1 and GBAP1. We used two sets of primers, as previously described [Sanger sequencing was performed for all individuals with a rare “pathogenic”/“likely pathogenic”/“uncertain” escribed . Individescribed , to deteGBA1 variants into true-positive, false-positive, and false-negative calls for each data analysis pipeline . True-positive variants were defined as GBA1 variants detected with Nanopore sequencing that were validated with Sanger sequencing. False-positive variants are those identified with Nanopore sequencing but not confirmed with Sanger sequencing. False-negative variants were determined as those not called with a specified data analysis pipeline, but later validated with Sanger sequencing. Evaluation of each data analysis pipeline was based on the ratio of false-positive to false-negative calls.To assess the performance of variant calling, we stratified the detected GBA1 single nucleotide variants were first filtered based on GnomAD frequency < 2%. Pathogenicity classification and scoring were assessed with American College of Medical Genetics and Genomics (ACMG) [GBA1 variants were categorized as “pathogenic”, “likely pathogenic”, or of “uncertain significance” and further validated by Sanger sequencing. The p.E365K variant, which is a known risk factor for PD, was additionally categorized as of “uncertain significance”, despite the “likely benign” score from ACMG. Variants categorized as of “uncertain significance” without information from mutation predictors were not Sanger sequenced.s (ACMG)  criterias (ACMG) , ClinVars (ACMG) , SIFT [4s (ACMG) , Polyphes (ACMG) , CADD [4s (ACMG) , and GERs (ACMG) . GBA1 vaGBA1 variants to the frequencies of GBA1 variants in controls, odds ratios were calculated. For this, the number of patients with PD carrying a GBA1 variant was multiplied by the number of controls without a GBA1 variant and subsequently divided by the number of patients with PD without a GBA1 variant that was multiplied by the number of controls carrying a GBA1 variant.To compare the number of Norwegian PD patients with and without GBA1 variants detected in PD and the frequency across different populations , articles that did not perform GBA1 variant screening or examine GBA1 variant frequency in their study population (n = 22), and articles that did not include patients with PD (n = 10). Multiple reasons for exclusion were possible. In addition to the articles found via the search term, suitable articles that were referenced in this literature were also included in the overview.We performed a systematic literature review to summarize Below is the link to the electronic supplementary material.Supplementary Material 1


Acinetobacter baumannii is recognised as one of the most significant antibiotic-resistant bacteria, capable of carrying up to 45 different resistance genes, and new drug discovery targets for this organism is an urgent priority. Short-chain dehydrogenase/reductase enzymes are a large protein family with >60,000 members involved in numerous biosynthesis pathways. Here, we determined the structure of an SDR protein from A. baumannii and assessed the putative co-factor comparisons with previously co-crystalised enzymes and cofactors. This study provides a basis for future studies to examine these potential co-factors in vitro.Bacterial antibiotic resistance remains an ever-increasing worldwide problem, requiring new approaches and enzyme targets.  Acinetobacter baumannii is an important drug-resistant pathogen, with infection rates of ~2% of all healthcare-associated infections in the United States and Europe and ~4% in the Middle East and Asia in 2017 [A. baumannii exhibits high rates of multidrug resistance (MDR), that is ~45% globally and up to 70% in Latin America and the Middle East [A. baumannii was listed as a priority one, critical pathogen [Antimicrobial resistance (AMR) is arguably one of the most critical challenges worldwide, with predictions estimating AMR infections to be a leading cause of mortality by 2050 without new treatment alternatives . Acinetodle East . The WorAcinetobacter baumanni, a gram-negative, non-motile, pleomorphic bacillus, is an opportunist and robust species with a remarkable ability to survive, adapt, and accumulate AMR. The capacity for AMR in A. baumanni stems from both natural and acquired features. A. baumannii has a minimal number of small porins in the outer membrane, resulting in a low permeability to antibiotics [A. baumanni the ability to utilise different AMR enzymes such as AmpC cephalosporinases, OXA carbapenemases, KPC serine carbapenemases, metallo-carbapenemases, and aminoglycosides, and form the basis for an extremely drug resistant pathogen [ibiotics . The bacibiotics , and haribiotics ,7. Moreoibiotics . These gpathogen .A. baumannii has the capacity for both nosocomial and community-acquired infections leading to meningitis, septicaemia, pneumonia, wound and urinary infections, with increased risk through the use of catheters and ventilators in immunocompromised and elderly patients [Acinetobacter in hospitals due to the ability of this bacteria to adhere to plastic and form biofilms [A. baumannii [A. baumannii [A. baumannii community-acquired infections including acute pneumonia [Within a clinical setting, patients . Endotrabiofilms . Additioaumannii . Postsuraumannii –14. Commaumannii ,16. Undeneumonia .A. baumannii has displayed resistance and reduced sensitivity to antimicrobials including last resort carbapenem, polymyxins, and tigecyclines [A. baumannii (Uniprot ID: A0A0M3FP68) was cloned, expressed, purified, and crystallised. Crystals diffracted to 2.4 Å, permitting structure determination. Based on the structure and bioinformatics analysis, potential cofactors were investigated.cyclines . Due to cyclines ,19,20. Tcyclines  carry oucyclines . SDR famcyclines . Here, aA. baumannii was codon-optimized, cloned into a pMCSG21 expression vector, expressed using auto-induction method, and purified as previously described [E. coli BL21(DE3) cells were transfomed with plasmid using the heat shock method. Protein expression was carried out for 36 h at room temperature, and cells were harvested by centrifugation. Following two freeze-thaw cycles, cells were treated with DNase (∼0.025mg/mL) and lysozyme (∼2mg/mL). After centrifugation, the soluble extract was injected onto a pre-equilibrated GE HisTrap 5mL column, washed with 10 column volumes of His A buffer  and eluted using His B buffer . Fractions containing protein were pooled and treated with tobacco etch virus protease at 4°C for ~14h. Size exclusion chromatography was performed using Superdex 200pg 26/600 column using Tris-Buffered Saline . Purified protein was concentrated and stored at -80°C for future assays [The putative SDR sequence (Uniprot ID: A0A0M3FP68) from escribed . In summNase ∼0.0mg/mL ande assays , merged e assays , phased e assays , refinede assays . To compe assays , and thee assays . The PDBe assays .A. baumannii has been identified as a critical pathogen in desperate need of new treatment options due to the alarming rate of increasing resistance to all classes of antibiotics, including last resort carbapenem [A. baumannii SDR enzyme with no known structure, and for which the closest sequence identity to any known protein structure in the Protein Data Bank is ~37% to expand the repertoire of previously determined SDR proteins from this important bacterium [1 with iMosflm and scaled and merged in Aimless to 2.4 Å . Four molecules were placed in the asymmetric unit, and rebuilding/refinement was performed to an R/Rfree of 19.0 and 22.6% respectively. Due to poor density, residues 49–53 and 189–196 of chains A and C, and residues 50–51 and 189–197 of chains B and D were unable to be modelled, and density for the N-terminal methionine was not of sufficient quality to model this residue. After structural elucidation it was discovered that Gln69 was missing, and investigation revealed this error was introduced in the original design of the plasmid. Fortunately, overlay of the experimentally determined structure with the Gln69 deletion and the alpha fold prediction without the Gln69 deletion showed that the overall structure was not affected, and it overlayed perfectly with previously determined SDR structures. Each monomer contains a central β -sheet (β1- β7), sandwiched between two groups of α- helices (α1-α2-α12-α13 and α3-α5-α6-α7-α8-α9-α10), with an overall topology of β1-α1-β2-α2-β3-α3-β4-α4-α5-α6-α7-β5-α8-α9-α10-β6-α11-α12-α13-β7-α14 /1407.7(B) Å2. The A/D interface was mediated by 11 hydrogen bonds and 2 salt bridges and buried an interface of 914.4(A)/906.5(B) Å2.To examine key interactions that mediate binding at the tetramer interface, the crystal structure was analysed using Proteins, Interfaces, Structures and Assemblies (PISA) ,39. Threin vitro due to crystallographic packing or other forms of regulation [T. thermophilis species (green) and a hydroxysteroid dehydrogenase from Homo sapiens (red) . In the Homo sapiens SDR structure Arg19, Ile21, Asp40, Asp62, Val63, Asn89, Ile187, Thr189 hydrogen bonded with NAD, of which three were conserved in the SDR of this study . All four catalytic residues, previously characterised to have catalytic function in SDRs, are conserved in the SDR structure determined in this study   . The res Lys153) .The second comparison was performed between the SDR of this study (dark blue) and SDR enzymes containing the NADPH cofactor; PDB 4BMS in yellow; PDB 4I5E in orange; PDB 6IHI in cyan and PDB 4JRO in purple Click here for additional data file.


However, steatosis is a growing pathology caused by the accumulation of lipids in hepatic cells due to increased lipogenesis, dysregulated lipid metabolism, and/or reduced lipolysis. Accordingly, this investigation hypothesizes a selective in vitro accumulation of palmitic and linoleic fatty acids on hepatocytes. After assessing the metabolic inhibition, apoptotic effect, and reactive oxygen species (ROS) generation by linoleic (LA) and palmitic (PA) fatty acids, HepG2 cells were exposed to different ratios of LA and PA to study the lipid accumulation using the lipophilic dye Oil Red O. Lipidomic studies were also carried out after lipid isolation. Results revealed that LA was highly accumulated and induced ROS production when compared to PA. Lipid profile modifications were observed after LA:PA 1:1 (v/v) exposure, which led to a four-fold increase in triglycerides (TGs) (mainly in linoleic acid-containing species), as well as a increase in cholesterol and polyunsaturated fatty acids (PUFA) content when compared to the control cells. The present work highlights the importance of balancing both PA and LA fatty acids concentrations in HepG2 cells to maintain normal levels of free fatty acids (FFAs), cholesterol, and TGs and to minimize some of the observed in vitro effects  caused by these fatty acids.Lipid metabolism pathways such as  In 2019, the worldwide incidence of non-alcoholic fatty liver disease (NAFLD) was one hundred and seventy million, corresponding to twice the number of cases registered in 1990 . The afoHigher free fatty acid (FFA) uptake, increased hepatic lipid accumulation, de novo lipogenesis, and endoplasmic reticulum (ER) stress are hallmarks of NAFLD development and progression ,8,9. OncLipid droplets are cellular organelles that originate from the ER and consist of a core of neutral lipids (cholesterol esters and triglycerides) enveloped by phospholipids and proteins . These sAs the accumulation of lipids in hepatocytes is considered a pathologic hallmark, in vitro models of steatosis have been used to study the hepatocellular consequences of lipid accumulation in human hepatic cells. These models include hepatocyte cell lines and primary hepatocytes treated in culture with monounsaturated and saturated fatty acids in order to mimic and explore novel features of NAFLD .Considering that Western diets are characterized by high intake of animal fats containing palmitic acid (PA) and seed oils with high linoleic acid (LA), this study aims to determine the effect and accumulation of PA and LA on hepatocytes ,18,19. Tv/v) of these FFA.To understand the interference of each fatty acid, saturated  and polyunsaturated (linoleic acid—LA), in the hepatocyte metabolic inhibition, the PrestoBlue assay was performed. v/v) ratio (approx. 100% vs. 20% in Control Cells). On the other hand, lower concentrations or the absence of LA in the 1 mM FFA mixture  were associated with lower metabolic inhibition values, namely, 1.54%, −50.01%, and −15.41%, respectively. These results indicate that LA induces higher metabolic inhibition than PA. However, such behavior may also be related to the proportion of dead, apoptotic, and live cells.As shown in v/v) are shown in v/v), 68.46 ± 0.62% for 1:1 (v/v), and 64.98 ± 6.12% for 0:1 (v/v).To evaluate if the metabolic inhibition results were in accordance with cell death, the apoptosis stages detection kit was used. The obtained percentage of live, early apoptotic, necrotic, and dead cells after the exposure to 1 mM LA:PA at different ratios , 0.88 ± 0.19% for 1:1 (v/v), and 2.59 ± 0.30% for 0:1 (v/v) (p > 0.05). Moreover, LA (1 mM) exposure resulted in a higher level of necrotic cells (26.89 ± 3.92%) compared to the other assayed conditions (p < 0.05). Interestingly, the percentage of dead cells was higher when PA was in the mixture. The 0:1 (v/v) ratio, containing 1 mM of PA, significantly led to the highest percentage of cell death (22.85 ± 3.63%) when compared to control.For early apoptosis, the percentages of live cells were: 1.40 ± 0.68% for Control, 0.73 ± 0.27% for LA:PA at 1:0  was evaluated to confirm if LA accumulation result in higher oxidative stress than PA in HepG2 cells. The results of relative fluorescence units (RFU) after exposure of hepatocytes to 2′,7′-dichlorofluorescein diacetate (DCFDA) for 5 h are depicted in v/v) showed an increase (p < 0.05) in ROS production after 2 h of DCFDA probe incubation when compared to the other ratios. Moreover, after 3 to 5 h of probe incubation, the presence of PA (LA:PA 1:1 and LA:PA 0:1 v/v) showed a higher (p < 0.05) RFU value when compared to the control. Hence, the mixture of 1 mM LA:PA 1:1 (v/v) resulted in a similar RFU value as the presence of LA:PA at 0:1 (v/v) (p < 0.05). After 5 h, superior ROS production and, therefore, higher RFU values, were achieved (p < 0.05) with the presence of LA , which was approximately the double of the RFU value obtained for LA:PA 1:1 and LA:PA 0:1 (v/v), 8653 and 7180 RFU respectively. The reported results are in accordance with the hypothesis that, due to the presence of double bonds, LA is more prone to oxidation than PA on HepG2 cells.Depending on the nature of the FFA, different levels of ROS were observed. Cells exposed only to LA  were of great relevance to discriminate some cellular effects of LA, PA, as well as the mixture of both FFA at equal concentrations. Besides these findings, and because PA is mostly associated with higher risk of cardiovascular diseases and as LA is an essential fatty acid ,21, theiv/v) at 1 and 2 mM are not statistically different (p > 0.05), which indicates a low accumulation of PA independently of the assayed concentration (1 and 2 mM).Changes in cellular metabolism and lipid accumulation are increased by NAFLD and in vitro studies using ORO lipophilic dyes are usually used to predict lipid presence . Hence, v/v) ratio was selected to study the lipid profile of HepG2 cells, as this ratio increased lipid accumulation, as well as reactive oxygen species without metabolic inhibition impairment.Based on the previously discussed results, the LA:PA 1:1 (The fatty acids (FA) content (mg FA/g cell pellet) obtained by GC with flame-ionization detector (GC-FID) is shown in v/v) indicates, in general, a significant accumulation in terms of total FAs . Moreover, the FA profile showed that the LA was at lower concentrations (p < 0.01) in the control when compared to the exposed cells . Overall, PA and LA were the main fatty acids present but, after exposure, the PA concentration (60.77 ± 0.12 mg FA/g cell pellet) was approximately half of the LA concentration value .The analysis of the total FA (free and esterified) profile, observed in the cell pellet after exposure to LA:PA 1:1 , cis-11,14-eicosadienoic (C20:2c11c14), cis-8,11,14-eicosatrienoic (C20:3c8c11c14), and arachidonic acid (C20:4c5c8c11c14) were other detected FAs with significantly increased concentrations when cells were cultured in the presence of the FFA mixture.The results confirm that LA and PA are not equally accumulated and correlate with the previous observations of this research work concerning ORO staining results .v/v)], data suggest that LA was highly accumulated. Thus, the following GC-MS analyses were performed to understand if this accumulation was as FFA or TG.The performed FA analysis . allowedv/v)/control cells] of the free PA, LA, oleic and stearic acids, as well as cholesterol and triglyceride content, can be seen in The fold-change variation [LA:PA 1:1 (v/v)] ] II were cv/v)] .v/v) exposure increased intracellular TG accumulation. The analysis of the TG species was performed by LC-ESI-qTOF and the isotopic pattern of the molecular ion, as well as its MS2 fragmentation, which allowed TG identification by comparison with the reference spectra (used database: MS-Finder v3.52 and Lipid Maps®).Previous results revealed that 1 mM LA:PA at 1:1 (p < 0.05) were detected, and data suggested that there was a predominance of LA in the structure of the identified TGs. In general, PA was also present, but only in two of the identified species and in one of the sn positions . Thus, results revealed that the presence of TG C18:2, C18:2, C18:2; TG C16:0, C18:2, C18:2 and TG C16:0, C18:1, C18:2 was an effect from the LA and PA exposure and further accumulation.As can be seen in Fatty acids such as linoleic acid (LA) and palmitic acid (PA) have different physical and biological properties. Thus, LA is an essential fatty acid, and PA is the precursor of long chain fatty acids, such as oleic acid ,20. The Additionally, a two-fold increase in cholesterol was observed. This increase can contribute to cell damage since it is established that free cholesterol is hepatotoxic, inducing inflammation and fibrosis ,18.The hepatocyte protection mechanism linked to lipo-toxicity of FFA  is generAkazawa et al. 2018) described that the predominance of saturated FFAs in hepatocytes can decrease cellular membrane fluidity and induce ER stress, causing cell death . On the  describeFor culture assays, Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), and penicillin–streptomycin antibiotic were purchased from Thermofischer, Waltham, MA, USA. HepG2 (HB-8065™) cells were purchased from American Type Culture Collection (ATCC). PrestoBlue . Fatty Acid Free BSA was purchased from Merck . Dimethyl sulfoxide was purchased (DMSO) ). For total cellular protein quantification, Pierce™ BCA Protein Assay Kit was purchased from Thermofischer .Palmitic Acid/Hexadecanoic Acid (>99%) was purchased from Larodan , and linoleic acid (>99%) was purchased from Merck .For GC-MS analysis, all the samples were derivatized with N, O-Bis(trimethylsilyl) trifluoroacetamide with 1% trimethylchlorosilane (BSTFA), purchased from Merck . Regarding LC-ESI-qTOF analysis, all reagents were LC-MS grade, and isopropanol (IPA), acetonitrile (ACN), formic acid, and ammonium formate were purchased from VWR .For HPLC-ELSD analysis, chloroform  was purchased from Thermofischer , tetrahydrofuran (THF)  was purchased from Merck , ultra-pure water was obtained through a Milli-Q system , and acetic acid (HPLC grade) was obtained from Carlo Erba Reagents .The reagents used for lipid extraction were obtained as follows: dichloromethane (DCM)  and n-Hexane (≥97%) from VWR Chemicals , Methanol  from Honeywell , sodium methoxide  from Thermofischer , sulfuric acid (95.0–97.0%) from Honeywell , and dimethylformamide (DMF)  from Thermofischer .v/v) and then filtered using 0.20 µm-pore size membrane.Stock solutions of each fatty acid were prepared at 20 mM in DMEM with 1% of fatty acid free BSA. For PA, the stock solution was heated at 75 °C for 10 min and a further 15 min on a Bandelin Sonorex Super RK 106 Ultrasonic Bath  to improve dissolution. Fatty acids mixture was diluted to 1 mM final concentration using different ratios of LA:PA  supplemented with 10% of FBS and 1% of Penicillin-streptomycin Antibiotic at 37 °C, with 5% of CO4 cells/well in 96-well plates and then exposed to FFA diluted in DMEM with 0.1% fatty acid free BSA for 24 h, in quadruplicates. Wells with media supplemented with LA and PA (without cells) were used to subtract a possible influence of the samples in the PrestoBlue fluorescence signal. Cells treated with 10% DMSO were used as negative control. After incubation, PrestoBlue reagent was added to the media and incubated for 2 h. The fluorescence signal was read in a Synergy H1 microplate reader . Results were expressed in percentage of metabolic inhibition in comparison to cells without treatment. At least two independent experiments were performed.Metabolic inhibition of lipidic extracts on HepG2 were evaluated using PrestoBlue assay according to manufacturer’s instructions. Cells were seeded at 1 × 105 cells/well in 24-well plates and then exposed to FFA for 24 h, in duplicates. After 24 h, cells were washed twice using phosphate-buffered saline (PBS), trypsinized, and washed again with PBS twice by centrifugation . To perform the staining procedure, fluorochrome-labeled Annexin V was used to precisely target apoptotic cells. The apoptotic and necrotic events of cells treated with the different fatty acids (LA and PA) were determined using the Biolegend FITC Annexin V Apoptosis Detection Kit with 7-AAD Cat. 640922 , according to the manufacturer’s instructions and analyzed by flow cytometry using a BD Accuri™ C6 Plus Flow Cytometer .Cells were seeded at 3 × 10− 7AAD−), early apoptotic (Av+ 7AAD−), necrotic (Av− 7AAD+), and dead (Av+ 7AAD+) cells.Results are expressed in percentage of live (Av5 cells/mL and added to each well 100 μL of the appropriate sample (FFA mixture) solutions and incubated during 24 h. Afterwards, the DCFDA probe was prepared and added to the final concentration of 25 μM to each well. The fluorescence of the dye was measured immediately (0 h) using Synergy H1 microplate reader  and for 1, 2, 3, 4 and 5 h of probe incubation at 495/529 nm (Ex./Em.) .The intracellular generation of ROS in HepG2 cells was determined using the 2′,7′-dichlorofluorescein diacetate (DCFDA)  probe. Before the FFA exposure, cells were seeded in black, Thermofisher 96 clear-bottom well plate  at 1 × 105 cells/mL (1 mL) in 24-well plates for 24 h and then treated with the FFAs mixtures (500 µL of mixture per well). After 24 h of exposition, cells were washed twice using PBS (500 µL per well) and fixed in 4% Paraformaldehyde (PFA) diluted in PBS for 30 min to 1 h.The neutral lipid content accumulation was evaluated according to the method described by Forbes-Hernández et al. (2017) , with soOil Red O (ORO) stock solution was prepared in 2-propanol at 3.5 mg/mL and filtered through a 0.45 μm pore size hydrophobic membrane. Then, a working solution was set up using three parts of the ORO stock solution and two parts of water. Fatty acid treated and control cells were stained with Oil Red O working solution for 15 min at room temperature (250 µL per well), and PBS was later used to remove the non-stained ORO by washing wells three consecutively times.Afterwards, cell images were taken using ZEISS  Optical Microscope Axiocam 208 Color Camera. For semi-quantitative analysis of the stained lipids, 250 µL per well of 2-propanol was added to the 24-well plate containing cells, and then the solution was transferred to a 96-well microplate, and the absorbance was measured at 510 nm in Synergy H1 microplate reader . The absorbance was normalized by cellular protein quantification at 562 nm using Pierce™ BCA Protein Assay Kit.After 24 h of fatty-acid exposure, cells were observed, and then the supplemented fatty acid medium was aspired, and cells were once washed with PBS. Cells were trypsinized using TrypLE for 10 min and transferred to a falcon tube to be centrifuged . The obtained pellet was collected, and PBS was added and centrifuged , and the supernatant discarded afterwards.The total lipid content of each pellet was acquired using methyl-tert-butyl ether (MTBE) and methanol according to Matyash et al. (2008) extraction procedure .2O  (Phase A) and IPA:ACN  (Phase B), both containing 0.1% (v/v) of formic acid and 10 mM ammonium formate modifiers. Gradient of B phase flow was as follows: 0.0 min—40%; 2.0 min—43%; 2.1 min—50%; 12.0 min—54%; 12.1 min—70%; 18.0 min—99%; 20.0 min—99%; 20.1 min—40%; 22 min—40%. The flow rate was set at 0.4 mL/min, and column temperature was set at 55 °C. Injection volume was 3 µL in positive and 5 µL in negative ionization mode. For MS analysis, the following parameters were applied: end plate offset voltage 500 V, capillary voltage 4500 V (positive ionization) or 3000 V (negative ionization), nebulizing gas pressure of 35 psi, drying gas flow 8 L/min, drying gas temperature 325 °C, quadrupole ion energy 3 eV (positive ionization) or 5 eV (negative ionization), collision energy 10 eV (positive ionization) or 5 eV (negative ionization). Acquisition was performed in an Auto MS/MS scan mode over a mass range of m/z 50–1500. For both ionization modes, an external mass calibration was performed with a solution of IPA:upH2O  added with 0.2% (v/v) formic acid and 0.6% (v/v) NaOH 1 M, continuously injected at 180 µL/h. The acquired data were treated using MS-DIAL (v 4.90) and compound identification by MS-FINDER (v 3.52).Matyash cell extracts were prepared at a stock solution of 1 mg/mL and dissolved in IPA:ACN [9:1] at 0.20 mg/mL to be further analysed. Additionally, for quality requirements after data acquisition, a quality control (QC) sample was prepared using a pool of the samples and injected in positive mode on an UHPLC instrument  equipped with an Acquity UPLC BEH C18 (17 µm) pre-column , an Intensity Solo 2 C18 (100 × 2.1 mm) column , and coupled with an UHR–QTOF detector . Injection method was based on conditions reported by Sarafian et al. (2014)  and CaldThe Matyash extracts were analysed after derivatization into their trimethylsilyl derivatives. For that, to 1 mg of sample 70 µL of DCM and 60 µL of BSTFA were added. After 30 min incubation at 70 °C, DCM was added to a final volume of 400 µL. Afterwards, the samples were analysed on a GC-MS (triple quadrupole) model EVOQ  mass spectrometer, coupled with a Rxi-5Sil MS column (30 m × 250 µm × 0.25 µm) at constant flow of 1 mL/min. Helium was used as carrier gas, as described by Teixeira et al., (2022) . In summg; 18 °C; 5 min). The organic phase containing fatty acid methyl esters (FAME) was collected to a tared vial and then the hexane (approx. 1 mL) was evaporated and re-suspended in 300 uL of hexane for further analysis in a GC-FID apparatus equipped with a BPX70 column . The analysis conditions were set as follows: injector temperature 250 °C, split 25:1, injection volume 1 μL; detector (FID) temperature 275 °C; hydrogen was carrier gas at 20.5 psi; oven temperature program: started at 60 °C (held 5 min), then raised at 15 °C/min to 165 °C (held 1 min) and finally at 2 °C/min to 225 °C (held 2 min). The identification of fatty acids was carried out by comparison with a Supelco 37 FAME mixture sample (CRM47885).To determine the fatty acids profile, the performed method was based on Fontes at al. . The obtv/v) was injected in a high-performance liquid chromatograph (HPLC)  equipped with an evaporative light scattering detector (ELSD) and a Zorbax Eclipse Plus C8 column . The detector operation conditions were set with the evaporator at 30 °C, the nebulizer at 40 °C, and nitrogen as carrier gas with a flow rate set at 1.5 SLM. The mobile phases were prepared and filtered by a 0.20 µm-pore size hydrophobic membrane: A ) and B ) in a gradient mode were set between 0–45 min . The determination was carried out by injecting 10 µL of sample with a flow rate of 0.5 mL/min with the oven set at 40 °C.For the triglyceride analysis, the Plante et al. (2011)  methodolThe reported results showed that PA led to lower metabolic inhibition and, interestingly, when compared to LA, PA was poorly accumulated. Thus, LA was associated with higher accumulation in both FFA and TG forms. Triglyceride variation was analyzed and potential species identified, such as TG C18:2, C18:2, C18:2; TG C16:0, C18:2, C18:2 and TG C16:0, C18:1, C18:2, probably as a protective mechanism to limit FFA impact on cell damage.In Western diets, LA and PA sources are highly abundant ,18,19. TFuture work should study the binding and transmembrane transport of these FFA in HepG2 cells. The identification of other lipid species  involved in steatotic hepatocytes may also help to better understand and describe other underlying processes, as well as the study of the inflammatory and ER stress derived from the intracellular presence of LA and PA.


Subsequently, we successfully synthesized a cost-effective, magnetic CQDs/FeOx photocatalytic composite for the first time by combining the resulting CQDs and FeOx. Our findings demonstrated that calcining IOS in an air atmosphere enhanced the content of photocatalytically active α-Fe2O3, while incorporating WRN-based CQDs into FeOx improved the electron-hole pair separation, resulting in increased O2 reduction and H2O oxidation. Under optimized conditions , the CQDs/FeOx composite, utilizing WRN and IOS as its foundation, exhibited exceptional and reusable capabilities in photodegrading methylene blue and tetracycline. Remarkably, for methylene blue, it achieved an impressive degradation rate of 99.30% within 480 min, accompanied by a high degradation rate constant of 5.26 × 10−3 min−1. This composite demonstrated reusability potential for up to ten photocatalytic cycles without a significant reduction in the degradation efficiency, surpassing the performance of IOS and FeOx without CQDs. Notably, the composite exhibited strong magnetism with a saturation magnetization strength of 34.7 emu/g, which enables efficient and convenient recovery in photocatalytic applications. This characteristic is highly advantageous for the large-scale industrial utilization of photocatalytic water purification.To provide an economical magnetic photocatalyst and introduce an innovative approach for efficiently utilizing discarded waste rice noodle (WRN) and iron oxide scale (IOS), we initially converted WRN into carbon quantum dots (CQDs) using a hydrothermal method, simultaneously calcining IOS to obtain iron oxide (FeO Consequently, it has become essential to explore certain degrees of doping or composite modification to enhance their performance [3O4 nanoparticles to create CQDs/Fe3O4 composite material. This composite achieved an 83% degradation rate of alkaline methylene blue solution within 30 min [3N4 composite photocatalytic material, using FeCl3 as the iron source and citric acid and ethylenediamine as carbon sources. This composite achieved a photocatalytic rate of 99.3% for methylene blue within 120 min [In recent years, there has been growing interest in using semiconductor materials ,2,3 as p2 , ZnO [132 , CdS [152 ,17,18,192 [3O4 ,20, α-Fe3O4 [2O3 , β-Fe2O32O3 [2O3 , γ-Fe2O32O3 [2O3 , FeO [242O3 [2O4 )—have gaformance ,27,28. Aformance ,32,33,34n 30 min . Additio2 or ZnO to produce CQDs/TiO2 [2 or ZnO, particularly under visible light illumination, for a range of water-soluble dyes. This conversion strategy of WRN to CQDs/TiO2 [The improper disposal of cooking waste has the potential to introduce pollutants into the soil and water ecosystems, posing a considerable urban governance dilemma . The conQDs/TiO2  or CQDs/QDs/TiO2  photocatQDs/TiO2  or CQDs/QDs/TiO2  compositOn the other hand, iron oxide scale (IOS ) is formx). Subsequently, we combined the CQDs from WRN with the FeOx from IOS, achieving the pioneering synthesis of a cost-effective magnetic CQDs/FeOx photocatalytic composite material without the need for additional synthetic agents  and iron oxide scale (IOS), as the raw materials. Firstly, the WRN underwent a hydrothermal process to convert it into CQDs, while the IOS was calcined to obtain iron oxides (FeOents see . This ma2 (99% purity), 1,4-benzoquinone , 2-propanol , ethylenediaminetetraacetic acid disodium salt , as well as dimethyl pyridine N-oxide , were procured from Macklin Reagent  and were used as received, without undergoing additional purification steps.The waste rice noodle (WRN) was obtained from the canteen located at the Guilin University of Technology in Guilin, China, with its primary organic components including starch (21.36 g/100 g), protein (1.91 g/100 g), and fat (0.4 g/100 g). The waste iron oxide scale  was obtained from Valin Lianyuan Iron and Steel Co. Ltd. in Hunan, China. The tetracycline hydrochloride (98% purity), methylene blue (98.5% purity), nano ZnO (99% purity), nano TiOx photocatalytic composites, while the solid portion was separated to yield hydrothermal carbon (HTC) powder, which could be further treated to produce activated carbon [x as a brown powder.To initiate the process, the formulation of the CQDs solution commenced via a hydrothermal treatment applied to the WRN, adhering to the methodology outlined in our earlier research ,45. In td carbon . Simultax, choosing from three different amounts , in precisely measured proportions. Through meticulous magnetic stirring at room temperature for a duration of 0.5 h, a consistent and homogenous suspension was created. Subsequently, this well-mixed composition was placed into a 50 mL Teflon-lined autoclave, undergoing heating at a temperature of 85 °C for a span of 3 h to facilitate the formation of the composite structure. Following this process, the composite material was retrieved and subjected to centrifugation, followed by thorough triple washing with distilled water. Upon being subjected to vacuum drying at 60 °C, the resulting product emerged as the CQDs/FeOx photocatalytic composite material, taking the form of a deep brown powder. The samples were labeled as CQDs/FeOx–1 to CQDs/FeOx–7 based on the calcination temperature of the IOS and the amount of FeOx used. The relevant formulation design and elemental composition are presented in Next, a volume of 40 mL from the CQDs solution was skillfully combined with an appropriate quantity of FeOx composite material were conducted following the methods outlined in our previous reports [The characterizations and photocatalytic degradation experiments of the CQDs/FeO reports ,45. For x composite (CQDs/FeOx–3 sample), FeOx powder , and IOS  is depicted in 3O4, JCPDS card no. 19-0629). Additionally, the characteristic peaks at 33.1° and 35.6° could be attributed to the (104) and (110) crystal planes of hematite . This indicated that the IOS mainly contained Fe3O4 and α-Fe2O3, with Fe3O4 being dominant. Combining this with the results of the elemental composition analysis  was too small to exhibit an effective photocatalytic ability. As a result, the iron oxide photocatalytic component was mainly α-Fe2O3. Therefore, it was foreseeable that the waste IOS could not be directly used as a photocatalytic material or directly combined with CQDs due to the low content of α-Fe2O3, making calcination treatment necessary to enhance its photocatalytic potential.The PXRD analysis of the CQDs/FeOysis See , the Fe/x powders obtained through the calcination of IOS at 300 ℃ with that of IOS, it was evident that the diffraction peaks of the α-Fe2O3 were significantly enhanced after calcination. The characteristic peaks at 24.1°, 33.1°, 35.6°, 40.9°, 49.5°, and 54.1° corresponded to the (012), (104), (110), (113), (024), and (116) crystal planes of α-Fe2O3, respectively. Additionally, based on compositional analysis, the Fe/O mass ratio decreased to 2.51, indicating that the FeOx powder contained 43.3% of α-Fe2O3. This transformation implied that calcination in air could convert a portion of the initially non-photocatalytic Fe3O4 into photocatalytically active α-Fe2O3, which laid the foundation for obtaining practical photocatalytic materials after the composite with CQDs. The PXRD patterns of the CQDs/FeOx composite and FeOx exhibited resemblances; however, the diffraction peaks corresponding to the CQDs within the composite were not prominently discernible due to their relatively modest concentration.Comparing the PXRD pattern of the FeOx composite (CQDs/FeOx–3 sample) exhibited a Z-average particle size value of 127.8 ± 63.26 nm (refer to x composite (CQDs/FeOx–3 sample) are visually depicted in x composites exhibited an irregular lamellar structure, with uniformly dispersed spherical CQDs particles on the FeOx surface. In x and CQDs. The lattice stripes, exhibiting spacings of 0.185 nm, 0.198 nm, and 0.271 nm, corresponded, respectively, to the (104), (110), and (024) crystal planes of α-Fe2O3. Furthermore, discernible lattice stripes with spacings of 0.103 nm and 0.168 nm aligned with the (111) and (220) crystal planes of Fe3O4. These observations indicated that the obtained FeOx was primarily a mixture of magnetite (Fe3O4) and hematite (α-Fe2O3), which aligned with the PXRD results. Moreover, crystalline planes with a lattice spacing of about 0.283 nm, corresponding to the (020) crystalline planes of CQDs [x in the resulting composite.The results of the particle size distribution testing indicated that the obtained CQDs/FeOrefer to  in ESI.  of CQDs ,51, werex composite and FeOx powders are displayed in x composite. Notably, 1/2) and Fe(2p3/2) signals, respectively. A noteworthy separation of divalent and trivalent iron signals was discernible within the Fe(2p3/2) signal, indicative of the presence of both Fe2+ and Fe3+ ions in the resulting CQDs/FeOx composite. In the high-resolution spectrum of O 1s, the CQDs/FeOx complex displayed distinct Fe–O and C–O bonds, hosting characteristic signals positioned at 530.71 and 528.29 eV, respectively. In contrast, the O 1s spectrum of the FeOx calcined at 300 °C displayed two distinct peaks at 530.5 and 529.38 eV, attributing them to the Fe–O bond and the hydroxyl group present on the FeOx surface, respectively. This led to the inference that the interaction between CQDs and FeOx involved the carboxyl group within the CQDs and the Fe–OH group on the surface of the FeOx, leading to the vanishing of the surface hydroxyl signal and the emergence of the C–O bond signal, as indicated in x composite, the peak at 283.69 eV could be attributed to the C–C bond within the CQDs. Simultaneously, the signals centered at 285.03 and 287.51 eV corresponded to the C–O and C=C bonds of CQDs, respectively, confirming the successful integration of the CQDs and FeOx within the resultant composites.The XPS spectra of both the CQDs/FeOx exhibited an obvious Fe–OH stretching vibration peak at around 880 cm−1 in their IR spectra. However, this peak disappeared after the composite of FeOx with CQDs. Paired with the XPS findings, it is conceivable that the intricate interplay between the CQDs and FeOx constituted a response involving the carboxyl group within the CQDs and the hydroxyl group located on the FeOx surface. Furthermore, characteristic absorption peaks of CQDs were observed in the IR spectra of the CQDs/FeOx composites, including the stretching vibration (3422 cm−1) and bending vibration (1620 cm−1) of the O–H bond on the CQDs, and the stretching vibration (1063 cm−1) of the C–O bond, etc. In addition, due to the presence of organic groups such as hydroxyl and carboxyl groups in the CQDs, the PZC value of the CQDs/FeOx composites (8.46) was higher than that of the non-composite FeOx powder (7.98) (See x composite exhibited a relatively low specific surface area (2.28 m2 g−1) compared with the CQDs/TiO2 [x powder (1.57 m2 g−1) (See As shown in .98) See b. In addQDs/TiO2  and CQDsQDs/TiO2  photocatg−1) See c.x composite exhibited excellent photocatalytic degradation efficiency towards various organic pollutants. 3O4 component, the IOS itself lacked the ability for photocatalytic degradation to methylene blue, as its degradation rate was only 4.9% after 480 min of light irradiation, which was almost indistinguishable from the degradation without the catalyst. However, after calcination at 300 °C, a portion of the Fe3O4 in the IOS was converted into α-Fe2O3, resulting in the FeOx with a certain photocatalytic degradation ability. The degradation rate of the FeOx towards methylene blue reached 41.8% after 480 min of light irradiation, with an apparent degradation rate constant of 1.03 × 10−3 min−1. However, complete degradation could not be achieved under these conditions. In contrast, the catalytic effect of the CQDs/FeOx composite (CQDs/FeOx–3 sample) on methylene blue significantly improved after incorporating the WRN-based CQDs into the FeOx. The composite demonstrated a good photocatalytic degradation rate (up to 99.30% within 480 min) and a relatively high degradation rate constant (5.26 × 10−3 min−1), enabling the complete degradation of methylene blue  are shown in 2O3, even with the incorporation of CQDs, the catalytic effect could hardly be improved. For instance, the CQDs/FeOx–1 sample obtained by heating the IOS at 100 °C had a Fe/O mass ratio of 2.59 due to the relatively low calcination temperature. It was found that only about 10.9% of α-Fe2O3 was present in the mixed FeOx phase, similar to the IOS as a raw material. Consequently, despite containing CQDs, the photocatalytic degradation of CQDs/FeOx–1 remained poor. After 480 min of light exposure, its degradation rate to methylene blue was only 35.79%, and the degradation rate constant was merely 7.04 × 10−4 min−1, which is even worse than the FeOx sample without CQDs but calcined at 300 °C. There are reports indicating that, when heated in air at an appropriate temperature (around 300 °C), Fe3O4 exhibits a tendency to transform into α-Fe2O3 [x material showed a trend of initially decreasing and then increasing, indicating that the content of α-Fe2O3 first increased and then decreased. Among them, the CQDs/FeOx–3 sample prepared with IOS calcined at 300 °C had the smallest Fe/O mass ratio (2.51), which corresponded to the highest α-Fe2O3 content (44%). Therefore, it exhibited the best photocatalytic performance and achieved the complete degradation of methylene blue within 480 min, with a degradation rate constant of 5.26 × 10−3 min−1. Furthermore, the material demonstrated excellent recyclability, and its degradation rate remained above 98% even after ten cycles of photocatalysis  exhibited the best photocatalytic performance, with a degradation rate constant of 5.26 × 10−3 min−1 for methylene blue. However, it was found that further increasing the loading amount of CQDs may have an adverse effect on the photocatalytic performance. For example, in the case of the CQDs/FeOx–7 sample featuring a carbon content of 20.2 wt%, a comparatively reduced degradation rate constant of 3.73 × 10−3 min−1 was observed for methylene blue degradation. This phenomenon could be attributed to the potential shielding influence stemming from the presence of carbon-based constituents [Additionally, the loading amount of CQDs also affects the photocatalytic degradation capability of the CQDs/FeOtituents .x composite material could also be utilized for controlling antibiotic residues, as shown in x composite (CQDs/FeOx–3 sample) could degrade 98.21% of tetracycline within 320 min, with an apparent degradation rate constant of 3.73 × 10−3 min−1. In comparison, under the same conditions, the FeOx powder without CQDs could only degrade 40.45% of tetracycline, with an apparent degradation rate constant of 1.29 × 10−3 min−1.The obtained CQDs/FeOx composite reported in this paper demonstrated a comparably good photocatalytic performance. Moreover, due to the complete utilization of waste materials (IOS and WRN) as raw sources, the synthesis cost of this material was significantly reduced compared to other CQDs/iron oxide composites [2 and CQDs/ZnO composites based on WRN, the CQDs/FeOx composite exhibited a weaker photocatalytic performance  and residual magnetization intensity (5.8 emu/g). Therefore, the CQDs/FeOx composite still exhibits exceptional soft magnetic characteristics, making it a promising candidate for various magnetic applications.As mentioned earlier, after the process of calcination and compounding, the magnetic properties of the CQDs/FeOx composite material enable efficient and convenient recyclability in photocatalytic applications. Following the photocatalytic process, the CQDs/FeOx composite can be effortlessly separated using a magnetic field, achieving a mass recovery rate of 99.62% after the initial photocatalytic cycle and 98.45% after ten cycles of photocatalysis. Additionally, iron oxide photocatalysts have demonstrated excellent environmental compatibility, with minimal toxicity to fish and algae [x photocatalyst holds significant promise for large-scale industrial water purification.As depicted in nd algae ,57,58. Cx composite, the UV-VIS diffuse reflectance spectra of both the FeOx and the CQDs/FeOx composite are illustrated in x inorganic phase was in the ultraviolet region, with almost no absorption in the visible wavelength range above 380 nm. In contrast, the CQDs/FeOx composite material exhibited strong absorption throughout the entire visible light region, indicating that the introduction of CQDs enabled the composite material to utilize the energy in the visible light region more effectively, thereby generating more electron-hole pairs. Moreover, the determination of band gaps for the FeOx and CQDs/FeOx samples was performed through the application of the Tauc plot method [x was 2.17 eV, but after introducing CQDs, the band gap of the CQDs/FeOx composite material was further reduced to 1.40 eV. This narrower band gap allowed for more the effective utilization of energy in the visible light region, significantly promoting electron transitions and enhancing the photocatalytic degradation performance.To delve deeper into the photocatalytic mechanism of the resulting CQDs/FeOt method ,60, as dx composite and FeOx powder are determined using XPS valence spectra. As shown in 2O3, the VB potential of the FeOx powder was 2.59 eV, which was more positive than E0 (2.38 eV vs. NHE). This suggested that the FeOx powder could oxidize water to generate hydroxyl radical (·OH). However, the conduction band (CB) potential (ECB = EVB − Eg) of the FeOx was determined to be 0.42 eV, which was more positive than the standard electrode potential E0 for superoxide radicals (−0.046 eV vs. NHE). This indicated that the FeOx powder could not reduce oxygen in water to produce superoxide radical (O2·–). On the other hand, the introduction of CQDs raised the VB potential of the CQDs/FeOx composite material to 2.83 eV, enabling better oxidation of water to form photocatalytically active hydroxyl radical. Combining its own band gap (1.40 eV) and the band gap of CQDs (2.14 eV), the CB potential was calculated to be −0.08 eV, which was lower than E0 for superoxide radicals (−0.046 eV vs. NHE). This indicated that the CQDs/FeOx composite material could reduce oxygen in water to generate photocatalytically active superoxide radical.The valence band (VB) potentials of the CQDs/FeOx composite material and FeOx are presented in x composite material was notably subdued in comparison to that of the FeOx. This observation implies that the incorporation of CQDs could proficiently curtail the recombination of photogenerated electron-hole pairs, thereby significantly contributing to the enhancement of the photocatalytic degradation efficacy. The enhanced mechanism of the CQDs/FeOx composite material on the catalytic performance of the FeOx was also demonstrated through transient photocurrent response (PCR) under visible light irradiation and electrochemical impedance spectra (EIS). As shown in x composite material was approximately 11 times that of the FeOx without CQDs, indicating that the CQDs/FeOx composite could achieve a more efficient interface charge transfer and more effective electron-hole pair separation. The lower probability of photogenerated electron-hole recombination resulted in a significant improvement in the photocatalytic degradation performance. Additionally, the Nyquist plot of the CQDs/FeOx composite displayed a smaller semicircle diameter than that of the FeOx powder, indicating that the resulting CQDs/FeOx composite exhibited lower charge transfer resistance than the FeOx powder. This ensured a more efficient interface charge transfer and more effective electron-hole pair separation, consistent with the photocurrent analysis results  played a minor role and were not the primary drivers of the photocatalytic activity. In contrast, the incorporation of BQ or IPA substantially impeded the degradation efficiency, yielding photocatalytic efficiencies of 40.75% and 60.15%, respectively, as opposed to the scenario without quenching agents. This phenomenon implies that both superoxide radicals (O2·–) and hydroxyl radicals (·OH) stood as the predominant active species in the photocatalytic degradation mechanism, with O2·– playing a more pronounced role. Furthermore, using DMPO as a radical trapping agent, electron spin resonance spectroscopy (ESR) was carried out to study the active oxygen species generated by the CQDs/FeOx composite and FeOx. As shown in x composite resulted in strong characteristic peaks of both superoxide radical (O2·–) and hydroxyl radical (·OH). This indicates that the CQDs/FeOx composite material could reduce adsorbed O2 to form superoxide radical (O2·–) and oxidize adsorbed H2O to form hydroxyl radical (·OH) under light irradiation. In contrast, under the same test conditions, the FeOx powder could not generate superoxide radical (O2·–) effectively, while its signal of hydroxyl radicals was weak.The impacts of distinct quenching agents  on the photodegradation process of methylene blue are elucidated in x composite material was similar to that of the CQDs/TiO2 [x composite undergoes a dynamic process: the CQDs become readily excited by photogenerated electrons situated in the conduction band (CB), leaving behind holes in the valence band (VB). This excitation triggers rapid spatial electron transfer between the CQDs and FeOx particles, effectively suppressing recombination and yielding the enhanced separation of electron-hole pairs. As a result, photogenerated electrons amass in the CB of the CQDs, while the holes populate the VB of FeOx, with each entity primed for their distinct roles in photocatalytic reactions. Photogenerated holes engage with H2O to yield a profusion of ·OH radicals, while photogenerated electrons react with O2, leading to an abundant production of O2·– radicals. These generated O2·– and ·OH radicals collectively orchestrate the degradation of diverse organic pollutants, thereby showcasing exceptional prowess in the realm of photocatalytic degradation activity.As shown in QDs/TiO2  and CQDsQDs/TiO2  compositx composites by combining waste rice noodle (WRN) and iron oxide scale (IOS) and assessed their photocatalytic performance. The key steps in our synthesis strategy were identified: first, calcining IOS in an air atmosphere to enhance the photocatalytically active α-Fe2O3 content; second, incorporating WRN-based CQDs into FeOx to enhance the electron-hole pair separation, leading to increased O2 reduction and H2O oxidation. This significantly improved the photocatalytic performance. The resulting CQDs/FeOx composites efficiently degraded various organic pollutants under purple light irradiation and had superior magnetic properties, allowing for easy separation and reuse.In this study, we successfully synthesized low-cost CQDs/FeOx composite achieved the 100% conversion of WRN and IOS waste into high-value functional materials without additional synthetic additives. Compared to other reported photocatalytic composites, such as the CQDs/TiO2 and CQDs/ZnO from our previous work and other CQDs/iron oxide composites using synthetic reagents, our CQDs/FeOx composite is more cost-effective, environmentally friendly, and sustainable. Its magnetic properties enable rapid separation and efficient reuse, making it a promising candidate for large-scale, low-cost photocatalytic water purification with commercial potential. Furthermore, our approach to transform WRN and IOS into a CQDs/FeOx composite opens new possibilities for the combined utilization of waste.Our CQDs/FeO