Subclinical atherosclerosis and accelerated epigenetic age mediated by inflammation: a multi-omics study.

AIMS: Epigenetic age is emerging as a personalized and accurate predictor of biological age. The aim of this article is to assess the association of subclinical atherosclerosis with accelerated epigenetic age and to investigate the underlying mechanisms mediating this association. METHODS AND RESULTS: Whole blood methylomics, transcriptomics, and plasma proteomics were obtained for 391 participants of the Progression of Early Subclinical Atherosclerosis study. Epigenetic age was calculated from methylomics data for each participant. Its divergence from chronological age is termed epigenetic age acceleration. Subclinical atherosclerosis burden was estimated by multi-territory 2D/3D vascular ultrasound and by coronary artery calcification. In healthy individuals, the presence, extension, and progression of subclinical atherosclerosis were associated with a significant acceleration of the Grim epigenetic age, a predictor of health and lifespan, regardless of traditional cardiovascular risk factors. Individuals with an accelerated Grim epigenetic age were characterized by an increased systemic inflammation and associated with a score of low-grade, chronic inflammation. Mediation analysis using transcriptomics and proteomics data revealed key pro-inflammatory pathways (IL6, Inflammasome, and IL10) and genes (IL1B, OSM, TLR5, and CD14) mediating the association between subclinical atherosclerosis and epigenetic age acceleration. CONCLUSION: The presence, extension, and progression of subclinical atherosclerosis in middle-aged asymptomatic individuals are associated with an acceleration in the Grim epigenetic age. Mediation analysis using transcriptomics and proteomics data suggests a key role of systemic inflammation in this association, reinforcing the relevance of interventions on inflammation to prevent cardiovascular disease.

Microcrystalline Luminescent (Eu1-xLnx)2bdc3·nH2O (Ln = La, Gd, Lu) Antenna MOFs: Effect of Dopant Content on Structure, Particle Morphology, and Luminescent Properties.

In this work, three series of micro-sized heterometallic europium-containing terephthalate MOFs, (Eu1-xLnx)2bdc3·nH2O (Ln = La, Gd, Lu), are synthesized via an ultrasound-assisted method in an aqueous medium. La3+ and Gd3+-doped terephthalates are isostructural to Eu2bdc3·4H2O. Lu3+-doped compounds are isostructural to Eu2bdc3·4H2O with Lu contents lower than 95 at.%. The compounds that are isostructural to Lu2bdc3·2.5H2O are formed at higher Lu3+ concentrations for the (Eu1-xLux)2bdc3·nH2O series. All materials consist of micrometer-sized particles. The particle shape is determined by the crystalline phase. All the synthesized samples demonstrate an "antenna" effect: a bright-red emission corresponding to the 5D0-7FJ transitions of Eu3+ ions is observed upon 310 nm excitation into the singlet electronic excited state of terephthalate ions. The fine structure of the emission spectra is determined by the crystalline phase due to the different local symmetries of the Eu3+ ions in the different kinds of crystalline structures. The photoluminescence quantum yield and 5D0 excited state lifetime of Eu3+ are equal to 11 ± 2% and 0.44 ± 0.01 ms, respectively, for the Ln2bdc3·4H2O structures. For the (Eu1-xLux)2bdc3·2.5H2O compounds, significant increases in the photoluminescence quantum yield and 5D0 excited state lifetime of Eu3+ are observed, reaching 23% and 1.62 ms, respectively.

Photochemical Reduction of Silver Nanoparticles on Diatoms.

In this work, the photochemical reduction method was used at 440 or 540 nm excitation wavelengths to optimize the deposition of silver nanoparticles on the diatom surface as a potential DNA biosensor. The as-synthesized nanocomposites were characterized by ultraviolet-visible spectroscopy (UV-Vis), Fourier transforms infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), scanning transmission electron microscopy (STEM), fluorescence microscopy, and Raman spectroscopy. Our results revealed a 5.5-fold enhancement in the fluorescence response of the nanocomposite irradiated at 440 nm with DNA. The enhanced sensitivity comes from the optical coupling of the guided-mode resonance of the diatoms and the localized surface plasmon of the silver nanoparticles interacting with the DNA. The advantage of this work involves the use of a low-cost green method to optimize the deposition of plasmonic nanoparticles on diatoms as an alternative fabrication method for fluorescent biosensors.

The parkinsonian LRRK2 R1441G mutation shows macroautophagy-mitophagy dysregulation concomitant with endoplasmic reticulum stress.

Autophagy is a mechanism responsible for the degradation of cellular components to maintain their homeostasis. However, autophagy is commonly altered and compromised in several diseases, including neurodegenerative disorders. Parkinson's disease (PD) can be considered a multifactorial disease because environmental factors, genetic factors, and aging are involved. Several genes are involved in PD pathology, among which the LRRK2 gene and its mutations, inherited in an autosomal dominant manner, are responsible for most genetic PD cases. The R1441G LRRK2 mutation is, after G2019S, the most important in PD pathogenesis. Our results demonstrate a relationship between the R1441G LRRK2 mutation and a mechanistic dysregulation of autophagy that compromises cell viability. This altered autophagy mechanism is associated with organellar stress including mitochondrial (which induces mitophagy) and endoplasmic reticulum (ER) stress, consistent with the fact that patients with this mutation are more vulnerable to toxins related to PD, such as MPP+.

Quantification of differential tissue biomarker responses to microplastic ingestion and plasticizer bioaccumulation in aquaculture reared sea bream Sparus aurata.

Marine aquaculture is considered a potential source of microplastics (MPs). MPs can induce oxidative stress and damage in marine species. In this study we evaluated the impact of MPs intake in the commercial fish, Sparus aurata, from aquaculture facilities and the antioxidant response associated to this MPs ingestion in caged specimens for 120 days. Sampling was carried out at the beginning of the study (T0), at 60 days (T60) and at 120 days (T120). At each sampling stage, gastrointestinal tract, blood, plasma, liver and muscle samples were obtained to analyse MPs intake (gastrointestinal tract), oxidative stress markers (blood, plasma and liver) and plasticizers bioaccumulation (muscle). Fish sampled at T60 presented the highest MPs intake and plasticizers accumulated in muscle over time, but with a different pattern according to type: bisphenols and phthalates. This indicates MPs ingestion induces a differential tissue response in S. aurata. Similarly, stress biomarkers presented a differential response throughout the study, depending on the analysed tissue. In the case of oxidative damage markers, for malondialdehyde (MDA) an increase throughout the study was observed both in liver and blood cells but with a progressive decrease in plasma. In the case of phase I detoxifying enzyme activities in liver, 7-ethoxyresorufin O-deethylase (EROD), 7-benzyloxy-4-[trifluoromethyl]-coumarin-O-debenzyloxylase (BFCOD) and carboxylesterases (CE), showed a comparable decrease at T60 with a slight recovery at T120. In contrast, glutathione-S-transferase (GST) activity was significantly enhanced at T60 compared to the other sampling stages. In conclusion, MPs ingestion occurs in aquaculture reared seabream where potentially associated plasticizers accumulate in the muscle and both could be responsible for plasma and liver oxidative stress damage and alterations on detoxifying biomarkers responses.

Assessing microplastic ingestion and occurrence of bisphenols and phthalates in bivalves, fish and holothurians from a Mediterranean marine protected area.

Microplastic (MP) ingestion, along with accumulated plasticizers such as bisphenol A (BPA), bisphenol F (BPF), and bisphenol S (BPS), and phthalates represented by diethyl phthalate (DEP), dibutyl phthalate (DBP) and bis (2-ethylhexyl) phthalate (DEHP), were quantified in bivalves, fish, and holothurians collected from a coastal pristine area at the western Mediterranean Sea. MP ingestion in sediment-feeders holothurians (mean value 12.67 ± 7.31 MPs/individual) was statistically higher than ingestion in bivalves and fish (mean 4.83 ± 5.35 and 3 ± 4.44 MPs/individual, respectively). The main ingested polymers were polyethylene, polypropylene, and polystyrene. The levels of BPS, BPF, and DEHP were highest in bivalves' soft tissue; BPA and DBP had the highest levels in the holothurians' muscle. In addition, the levels of all plasticizers assessed were lowest in fish muscle; only BPA levels in fish were higher than in bivalves, with intermediate values between those of bivalves and holothurians. This study provides data on exposure to MPs and plasticizers of different species inhabiting Cabrera Marine Protected Area (MPA) and highlights the differences in MP ingestion and levels of plasticizers between species with different ecological characteristics and feeding strategies.

Chitosan-collagen-hydroxyapatite membranes for tissue engineering.

Tissue engineering is growing in developing new technologies focused on providing effective solutions to degenerative pathologies that affect different types of connective tissues. The search for biocompatible, bioactive, biodegradable, and multifunctional materials has grown significantly in recent years. Chitosan, calcium phosphates collagen, and their combination as composite materials fulfill the required properties and could result in biostimulation for tissue regeneration. In the present work, the chitosan/collagen/hydroxyapatite membranes were prepared with different concentrations of collagen and hydroxyapatite. Cell adhesion was evaluated by MTS assay for two in vitro models. Additionally, cytotoxicity of the different membranes employing hemolysis of erythrocytes isolated from human blood was carried out. The structure of the membranes was analyzed by X-rays diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and thermal stability properties by thermogravimetric methods (TGA). The highest cell adhesion after 48 h was obtained for chitosan membranes with the highest hydroxyapatite and collagen content. All composite membranes showed good cell adhesion and low cytotoxicity, suggesting that these materials have a significant potential to be used as biomaterials for tissue engineering. Graphical abstract.

Assessing Matrix Solid-Phase Dispersion Extraction Strategies for Determining Bisphenols and Phthalates in Gilthead Sea Bream Samples.

Microplastics (MPs) and nanoplastics (NPs) are widely spread in the environment, generating significant concern due to their potential impact on environmental health. Marine species usually ingest plastic fragments, mistaking them for food. Many toxic compounds, such as plastic additives that are not chemically bound to the plastic matrix, can be released from MPs and NPs and reach humans via the food chain. This paper highlights the development and validation of a straightforward solid-liquid extraction clean-up procedure in combination with a matrix solid-phase dispersion method using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) detection, enabling facile, precise, and reliable identification and quantitation of a total of six bisphenols and phthalates in gilthead sea breams. Under the optimized conditions, the developed method showed good linearity (R2 > 0.993) for all target compounds. The recoveries obtained were between 70 and 92%. The relative standard deviations (RSDs) for reproducibility (inter-day) and repeatability (intra-day) were less than 9% and 10%, respectively. The limit of detection (LOD) and limit of quantification (LOQ) for the target compounds ranged from 0.11 to 0.68 µg/kg and from 0.37 to 2.28 µg/kg, respectively. A new, efficient extraction methodology for the determination of BPA, BPS, BPF, DBP, DEP, and DHEP in gilthead seabream has been optimized and validated.

Selective Extraction of Diazepam and Its Metabolites from Urine Samples by a Molecularly Imprinted Solid-Phase Extraction (MISPE) Method.

In this research, a molecularly imprinted polymer (MIP) was synthesized by precipitation polymerization using oxazepam (OZ) as a template molecule and was subsequently applied as a selective sorbent for the extraction of diazepam (DZP) and its metabolites in urine samples using an SPE cartridge. OZ, temazepam (TZ), nordiazepam (NZ) and DZP were analyzed in the final extracts by high-performance liquid chromatography with diode array detection (HPLC-DAD). The SPE extraction steps were optimized, and the evaluation of an imprinting factor was carried out. The selectivity of the method for OZ versus structurally related benzodiazepines (BZDs), such as bromazepam (BRZ), tetrazepam (TTZ) and halazepam (HZ), was investigated. Under the optimum conditions, the proposed methodology provided good linearity in the range of 10-1500 ng/mL, with limit of detection values between 13.5 and 21.1 ng/mL and recovery levels for DZP and its metabolites from 89.0 to 93.9% (RSD ≤ 8%) at a concentration level of 1000 ng/mL. The proposed method exhibited good selectivity, precision and accuracy and was applied to the analysis of urine samples from a real case of DZP intake.

Identification and morphological characterization of different types of plastic microparticles.

The knowledge of the polymeric composition of microplastics (MPs) is interesting because offers useful information on the resistance, durability, and degradability of these materials, also allowing progress in the control of this contamination. However, there is currently a lack of reliable standardized methods for the identification, and characterization of the plastic microparticles. This work uses different techniques in a complementary manner for the identification, and characterization of MPs that more frequently are found in the environment. A total of 10 types of plastics were collected (polystyrene (PS), polyethylene terephthalate (PETE), polyethylene (PE), high- and low-density polyethylene (HDPE and LDPE, respectively), polyvinyl chloride (PVC), polypropylene (PP), polytetrafluoroethylene (PTFE), Polyamide (PA, Nylon 6,6) and poly-carbonate (PC)) and their chemical identification were analyzed by reflectance-attenuated infrared (FTIR-ATR). Furthermore, the samples were observed using light microscopy, and scan-ning electron microscopy (SEM). Also, staining with 12 different dyes was performed to improve the identification of microplastics. The results of this study revealed that PETE, PE, HDPE and LDPE, whose SEM images exhibited smoothness and flat uniformity of their surface, were not (or less) susceptible to adsorb staining solutions while PP, PA, PVC, and PTFE, were capable of adsorbing the dye solutions.

Human post-mortem organotypic brain slice cultures: a tool to study pathomechanisms and test therapies.

Human brain experimental models recapitulating age- and disease-related characteristics are lacking. There is urgent need for human-specific tools that model the complex molecular and cellular interplay between different cell types to assess underlying disease mechanisms and test therapies. Here we present an adapted ex vivo organotypic slice culture method using human post-mortem brain tissue cultured at an air-liquid interface to also study brain white matter. We assessed whether these human post-mortem brain slices recapitulate the in vivo neuropathology and if they are suitable for pathophysiological, experimental and pre-clinical treatment development purposes, specifically regarding leukodystrophies. Human post-mortem brain tissue and cerebrospinal fluid were obtained from control, psychiatric and leukodystrophy donors. Slices were cultured up to six weeks, in culture medium with or without human cerebrospinal fluid. Human post-mortem organotypic brain slice cultures remained viable for at least six weeks ex vivo and maintained tissue structure and diversity of (neural) cell types. Supplementation with cerebrospinal fluid could improve slice recovery. Patient-derived organotypic slice cultures recapitulated and maintained known in vivo neuropathology. The cultures also showed physiologic multicellular responses to lysolecithin-induced demyelination ex vivo, indicating their suitability to study intrinsic repair mechanisms upon injury. The slice cultures were applicable for various experimental studies, as multi-electrode neuronal recordings. Finally, the cultures showed successful cell-type dependent transduction with gene therapy vectors. These human post-mortem organotypic brain slice cultures represent an adapted ex vivo model suitable for multifaceted studies of brain disease mechanisms, boosting translation from human ex vivo to in vivo. This model also allows for assessing potential treatment options, including gene therapy applications. Human post-mortem brain slice cultures are thus a valuable tool in preclinical research to study the pathomechanisms of a wide variety of brain diseases in living human tissue.

Micro-diffusely abnormal white matter: An early multiple sclerosis lesion phase with intensified myelin blistering.

OBJECTIVE: Multiple sclerosis (MS) is a chronic central nervous system disease whose white matter lesion origin remains debated. Recently, we reported subtle changes in the MS normal appearing white matter (NAWM), presenting with an increase in myelin blisters and myelin protein citrullination, which may recapitulate some of the prodromal degenerative processes involved in MS pathogenesis. Here, to clarify the relevance of these changes for subsequent MS myelin degeneration we explored their prevalence in WM regions characterized by subtly reduced myelination (dubbed as micro-diffusely abnormal white matter, mDAWM). METHODS: We used an in-depth (immuno)histochemistry approach in 27 MS donors with histological presence of mDAWM and 5 controls. An antibody panel against degenerative markers was combined and the presence of myelin/axonal aberrations was analyzed and compared with the NAWM from the same cases/slices/regions. RESULTS: mDAWM-defined areas exhibit ill-defined borders, no signs of Wallerian degeneration, and they associate with visible veins. Remarkably, such areas present with augmented myelin blister frequency, enhanced prevalence of polar myelin phospholipids, citrullination, and degradation of myelin basic protein (MBP) when compared with the NAWM. Furthermore, enhanced reactivity of microglia/macrophages against citrullinated MBP was also observed in this tissue. INTERPRETATION: We report a new histologically defined early phase in MS lesion formation, namely mDAWM, which lacks signs of Wallerian pathology. These results support the prelesional nature of the mDAWM. We conceptualize that evolution to pathologically evident lesions comprises the previously documented imbalance of axo-myelinic units (myelin blistering) leading to their degeneration and immune system activation by released myelin components.

Effect of the Electric Field on the Biomineralization of Collagen.

Collagen/hydroxyapatite hybrids are promising biomimetic materials that can replace or temporarily substitute bone tissues. The process of biomineralization was carried out through a double diffusion system. The methodological principle consisted in applying an electric field on the incubation medium to promote the opposite migration of ions into collagen membranes to form hydroxyapatite (HA) on the collagen membrane. Two physically separated solutions were used for the incubation medium, one rich in phosphate ions and the other in calcium ions, and their effects were evaluated against the traditional mineralization in Simulated Body Fluid (SBF). Pre-polarization of the organic membranes and the effect of incubation time on the biomineralization process were also assessed by FTIR and Raman spectroscopies.Our results demonstrated that the membrane pre-polarization significantly accelerated the mineralization process on collagen. On the other side, it was found that the application of the electric field influenced the collagen structure and its interactions with the mineral phase. The increment of the mineralization degree enhanced the photoluminescence properties of the collagen/HA materials, while the conductivity and the dielectric constant were reduced. These results might provide a useful approach for future applications in manufacturing biomimetic bone-like materials.

Enhanced Chitosan Photoluminescence by Incorporation of Lithium Perchlorate.

An improvement in chitosan film photoluminescence was observed after adding LiClO4. FTIR spectra, XPS, DFT calculations, and XRD measurements show an alteration of the H-bonds and an increase in the amorphous character of chitosan. PL spectra display a growth in intensity in the visible region along with the incorporation of lithium, signaling a possible rise in the population density of tail states and, consequently, better photon absorption, as observed from UV-vis measurements. A mechanism through aggregation-induced emission effect is proposed to explain the different results. Although this work establishes the relation between structural changes provoked by LiClO4 incorporation and luminescence in the case of chitosan, we expect that the same approach could be generalized to similar polymeric structures.

Characteristics of COPD smokers and effectiveness and safety of smoking cessation medications.

INTRODUCTION: Smoking is the main cause of chronic obstructive pulmonary disease (COPD), and smoking cessation is the only treatment shown to be effective in arresting the progression of COPD. Different epidemiological and population-based studies have shown smokers with COPD to have specific smoking characteristics that differentiate them from the rest of smokers and which complicate smoking cessation. The main objective of this study is to analyze the effectiveness and safety of drug treatments for smoking cessation in smokers with severe or very severe COPD. METHODS: Smokers with severe or very severe COPD (Global Initiative for Chronic Obstructive Lung Disease stages III and IV) received treatment for smoking cessation. The treatment program consisted of a combination of behavioral therapy and drug treatment. Patients were followed up at 1, 2, 4, 6, 8, 10, 12, 18, and 24 weeks after the quit date. RESULTS: Four hundred seventy-two patients were seen, 65% were male, and their mean age was 58.3 (9.8). They smoked an average of 29.7 (13.4) cigarettes/day, and their mean Fagerström test for nicotine dependence score was 7.4 (2.1). Continuous abstinence rate from 9 to 24 weeks (CAR 9-24) was 48.5%. According to type of treatment used, CAR 9-24 for nicotine replacement therapy (NRT), bupropion, and varenicline were 38.2%, 55.6%, and 58.3%, respectively. Varenicline was more effective than nicotine patches: 61% versus 44.1% (odds ratio: 1.98; 95% CI: 1.25-3.12; p = .003). NRT was the treatment producing the fewest adverse effects. The onset of psychiatric symptoms due to medication was rare and evenly distributed across groups. CONCLUSIONS: This study shows that smokers with severe or very severe COPD are predominantly males with a high degree of physical dependence upon nicotine. CAR 9-24 was 48.5%. Varenicline and bupropion yielded higher abstinence rates than NRT. Varenicline was more effective than nicotine patches: all types of treatments were safe.

Electrochemical Sensor for Hydrogen Peroxide Based on Prussian Blue Electrochemically Deposited at the TiO2-ZrO2-Doped Carbon Nanotube Glassy Carbon-Modified Electrode.

In this investigation, a hydrogen peroxide (H2O2) electrochemical sensor was evaluated. Prussian blue (PB) was electrodeposited at a glassy carbon (GC) electrode modified with titanium dioxide- and zirconia-doped functionalized carbon nanotubes (TiO2.ZrO2-fCNTs), obtaining the PB/TiO2.ZrO2-fCNTs/GC-modified electrode. The morphology and structure of the nanostructured material TiO2.ZrO2-fCNTs was characterized by transmission electron microscopy, the specific surface area was determined via Brunauer-Emmett-Teller, X-ray diffraction, thermogravimetric analysis, and Fourier transform infrared spectroscopy. The electrochemical properties were studied by cyclic voltammetry and chronoamperometry. Titania-zirconia nanoparticles (5.0 ± 2.0 nm) with an amorphous structure were directly synthesized on the fCNT walls, aged during periods of 20 days, obtaining a well-dispersed distribution with a high surface area. The results indicated that the TiO2.ZrO2-fCNT-nanostructured material exhibits good electrochemical properties and could be tunable by enhancing the modification conditions and method of synthesis. Covering of the nanotubes with TiO2-ZrO2 nanoparticles is one of the main factors that affected immobilization and sensitivity of the electrochemical biosensor. The electrode modified with TiO2-ZrO2 nanoparticles with the 20-day aging time was superior regarding its reversibility, electric communication, and high sensitivity and improves the immobilization of the PB at the electrode. The fabricated sensor was used in the detection of H2O2 in whey milk samples, presenting a linear relationship from 100 to 1,000 µmol L-1 between H2O2 concentration and the peak current, with a quantification limit (LQ) of 59.78 µmol L-1 and a detection limit (LD) of 17.93 µmol L-1.

Assessment of the impact of aquaculture facilities on transplanted mussels (Mytilus galloprovincialis): Integrating plasticizers and physiological analyses as a biomonitoring strategy.

The growing plastic production and its continuous use is a significant problem. In addition, aquaculture practices have experienced a considerable growth and plastic is widely used in these activities, hence plasticizers must be considered due to their potential ecotoxicological impacts on species. Mussels placed inside an Integrated Multi-Trophic Aquaculture (IMTA) system and at two control locations were employed to quantify the ingestion of anthropogenic particles and associated chemical plasticizers, such as bisphenol A (BPA) jointly to bisphenol F (BPF) and bisphenol S (BPS), and phthalates represented by diethyl phthalate (DEP), dibutyl phthalate (DBP) and bis(2-ethylhexyl) phthalate (DEHP). In addition, some metabolism and oxidative stress related parameters were measured in mussels' whole soft tissue. Anthropogenic particle ingestion of mussels increased over time at the three locations and the following order of abundance of pollutants was observed: BPA> BPF> DEHP> DBP> BPS> DEP. Even though no differences according to location were found for pollutants' occurrence, time trends were evidenced for BPA and DEHP. On the other hand, a location effect was observed for biomarkers with highest values detected in mussels located at the vicinities of the aquaculture facility. In addition, a reduced detoxification activity was observed over time parallel to BPA decrease.

Biological effects of olive oil phenolic compounds on mitochondria.

Phenolic compounds derived from olive oil have beneficial health properties against cancer, neurodegenerative, and metabolic diseases. Therefore, there are discrepancies in their impact on mitochondrial function that result in changes in oxidative capacity, mitochondrial respiration, and energetic demands. This review focuses on the versatile role of oleuropein, a potent antioxidant that regulates the AMPK/SIRT1/mTOR pathway to modulate autophagy/mitophagy and maintain metabolic homeostasis.

Loss of KEAP1 Causes an Accumulation of Nondegradative Organelles.

KEAP1 is a cytoplasmic protein that functions as an adaptor for the Cullin-3-based ubiquitin E3 ligase system, which regulates the degradation of many proteins, including NFE2L2/NRF2 and p62/SQSTM1. Loss of KEAP1 leads to an accumulation of protein ubiquitin aggregates and defective autophagy. To better understand the role of KEAP1 in the degradation machinery, we investigated whether Keap1 deficiency affects the endosome-lysosomal pathway. We used KEAP1-deficient mouse embryonic fibroblasts (MEFs) and combined Western blot analysis and fluorescence microscopy with fluorometric and pulse chase assays to analyze the levels of lysosomal-endosomal proteins, lysosomal function, and autophagy activity. We found that the loss of keap1 downregulated the protein levels and activity of the cathepsin D enzyme. Moreover, KEAP1 deficiency caused lysosomal alterations accompanied by an accumulation of autophagosomes. Our study demonstrates that KEAP1 deficiency increases nondegradative lysosomes and identifies a new role for KEAP1 in lysosomal function that may have therapeutic implications.