Amiodarone, a common antiarrhythmic drug, is known for its severe side effects, including pulmonary toxicity, which involves oxidative stress and apoptosis. Artemisinin, an antimalarial drug, has shown cytoprotective properties by inhibiting oxidative stress and apoptosis. This study investigated the protective effects of artemisinin against amiodarone-induced toxicity in human bronchial epithelial cells (BEAS-2B) and mouse models.

In vitro experiments revealed that amiodarone decreased cell viability, increased LDH release, ROS generation, caspase 3 activation, and apoptosis in BEAS-2B cells. Artemisinin counteracted these effects by upregulating p-AMPK, CaMKK2, Nrf2, and SOD1 protein levels, thereby protecting the cells from oxidative damage. The protective effect of artemisinin was diminished by the AMPK inhibitor Compound C or AMPKα knockdown. In vivo experiments demonstrated that artemisinin increased p-AMPK and Nrf2 protein levels in lung tissues, protecting against amiodarone-induced apoptosis and bronchial epithelial cell shedding in mice.

These findings suggest that artemisinin protects airway epithelial cells and lung tissue from amiodarone-induced oxidative stress and apoptosis through AMPK activation, offering potential new strategies for preventing and treating amiodarone-induced pulmonary toxicity.

Inflammatory cell infiltration and myofiber necrosis are pathological hallmarks of idiopathic inflammatory myopathies (IIM). Methyl 3,4-dihydroxybenzoate (MDHB) is a natural phenolic acid compound, renowned for its anti-inflammatory and antioxidant effects. In this study, we investigated its protective mechanisms targeting muscle fiber necrosis and macrophage pyroptosis by regulating oxidative stress in IIM. In the present study, we found that increased reactive oxygen species (ROS) level and decreased nuclear factor erythroid 2 related factor 2 (Nrf2) protein expression were shown on the muscle fibers of experimental autoimmune myositis (EAM). Receptor-interacting protein 1 (RIPK1) and receptor-interacting protein 3 (RIPK3) protein expression were elevated in EAM. In vitro, MDHB protected C2C12 cells and myotubes against H2O2-induced cell viability damage. MDHB decreased the levels of oxidative stress such as ROS, and mitochondrial superoxide (MitoSOX), and rescued mitochondrial membrane potential and ATP generation. MDHB inhibited necroptosis of the C2C12 cells and myotubes under H2O2 stimulation in a dose-dependent manner. Furthermore, MDHB suppressed lipopolysaccharide and nigericin-induced caspase-1 cleavage and interleukin (IL-1β) secretion, indicating suppression of macrophage pyroptosis in vitro. In vivo, treatment with MDHB suppressed EAM-induced muscle weakness and inflammation. MDHB decreased ROS accumulation, and increased Nrf2 and heme oxygenase-1 (HO-1) expression in EAM mice's muscles, thereby inhibiting necroptosis of inflamed muscle species and macrophage pyroptosis. In conclusion, we demonstrated that MDHB could be a novel therapy for IIM that alleviates inflammation, muscle fiber necroptosis, and macrophage pyroptosis by regulating the Nrf2/HO-1 pathway.

CD68 is a vital costimulatory molecule expressed on macrophages/microglia (M/Ms) and plays a critical role in their activation. By targeting this molecule, therapeutic interventions can potentially prevent the homing of M/Ms. to the lesion site. In this study, we developed a biomimetic nanoparticle-based system (siCD68/NPs) to deliver CD68 small interfering RNA (siCD68) more efficiently into M/Ms.Administration of siCD68/NPs was found to not only polarize M1 macrophages toward M2 phenotype, but also reduce the reactive oxygen species (ROS) levels in lipopolysaccharide (LPS) plus interferon-γ (IFN-γ) induced macrophages/microglia (M/Ms). Moreover, treatment with siCD68/NPs significantly extended the survival time in a mouse spinal cord injury (SCI) model.In summary, siCD68/NPs were found to activate an anti-neuroinflammatory immune response and reprogram the polarization of M/Ms., leading to a significant improvement in the recovery of spinal cord injury. This study contributes to the field of biomimetic nanoparticle-based therapies and offers novel insights into potential treatments for neuroinflammation-induced SCI.

Oxidative stress is a key factor in neurodegenerative diseases, particularly in Alzheimer's disease (AD) which is the leading cause of dementia. A hallmark of AD is the accumulation of amyloid β (Aβ) peptides, along with redox-active metal ions and heme cofactors, all of which significantly contribute to disease progression. When heme binds to Aβ, it can drive oxidative stress through two primary pathways: firstly, reduced high-spin ferrous heme-Aβ active sites may generate H2O2 through oxygen reduction, leading to the oxidation of biomolecules and lipid membranes; secondly, this H2O2 can react with the oxidised form of high-spin ferric heme-Aβ, initiating peroxidase-like activity that can catalyse the oxidation of neurotransmitters. These pathways converge at a crucial intermediate i.e. the heme-Aβ-peroxo complex, which serves as the final intermediate in the ROS cycle and the first in the peroxidase cycle. Although, we have previously characterized other intermediates in these pathways, compound 0 resulting from the reaction of a high-spin heme-Aβ species with peroxides has remained elusive due to its rapid hydrolysis in an aqueous environment. In this study, we report the oxidation of dopamine by peroxides catalyzed by ferric heme-Aβ species and successfully stabilised and characterized compound 0 of high-spin heme-Aβ in dimethylformamide, an organic aprotic solvent. This stabilization enables detection through stopped-flow, EPR and resonance Raman spectroscopy, thereby facilitating a deeper understanding of the oxidative stress caused by high-spin heme-Aβ.

Microglial intrinsic immune checkpoints are essential safeguards to maintain immune homeostasis by preventing microglial overactivation, a process that substantially influences neurological disorders such as autism spectrum disorder (ASD). MEF2C is a crucial immune checkpoint that regulates microglial activation, but the mechanism remains unclear. We found that MEF2C-deficient (MEF2C-/-) induced microglia-like cells (iMGLs) derived from human pluripotent stem cells (hPSCs) exhibited overactivation following lipopolysaccharide stimulation, mimicking patterns observed in various neuroinflammatory disorders. High-throughput screening identified BMS265246, a cyclin-dependent kinase 2 (CDK2) inhibitor, which suppressed overactivation of MEF2C-/- iMGLs and normalized their inflammatory responses. Mechanistically, MEF2C transcriptionally upregulated p21 to inhibit CDK2 activation-mediated retinoblastoma protein (RB) degradation, thereby preventing transcription factor nuclear factor κB (NFκB) nuclear translocation and consequent microglial overactivation. BMS265246 treatment substantially ameliorated microglial overactivation and ASD-like behaviors in Mef2c-deficient mice. Our findings identify the MEF2C-p21-CDK2-RB-NFκB axis as a critical pathway to maintain microglial homeostasis and highlight CDK2 as a potential therapeutic target for neuroinflammation.

Prevalent microcystin-LR (MC-LR), a cyanotoxin, in agricultural fields compromises produce safety and threatens human health. However, little is known about its uptake and accumulation in plant cells and its resultant toxicity mechanisms. This study revealed that the MC-LR uptake into protoplasts was controlled by an active transmembrane transport process mediated by the protein carrier. MC-LR in the plant cells can enlarge the specific mitochondrial permeability transition pores and probably bind with the electron transport chain complex I (especially, NADH oxidoreductase 1, -30.59 kcal/mol of binding energy) and complex III (especially, cytochrome b, -36.98 kcal/mol of binding energy) via hydrophobic force and hydrogen bond. The interactions between MC-LR and the mitochondrial complex proteins block the electron transfer, causing high levels of reactive oxygen species (ROS), especially for H2O2. The MC-LR-induced ROS destroys the mitochondrial inner membrane structure and decreases the cell viability by 13.6-30.6% in a significant dose-dependent manner at 1-5 mg/L MC-LR stress. The findings provided direct evidence of MC-LR entry into the cells via active plasma membrane transport for the first time and clarified the associations between MC accumulation and its toxicity at cellular and molecular levels, thereby providing crucial insights for ensuring food safety and safeguarding human health.

Neonatal hypoxia-ischemia (HI) is a major cause of mortality and neurological impairments in infants. Main HI-induced pathological mechanisms include mitochondrial dysfunction and oxidative stress due to insufficient oxygen and energetic substrates to the nervous cells. Bovine lactoferrin (Lf) has demonstrated neuroprotective effects in several experimental models of neonatal brain injury in rodents, however its mechanisms remain unclear. This study aimed to evaluate the early impact of maternal dietary supplementation with Lf on redox and hippocampal mitochondrial function following neonatal HI. From postnatal day 6 (PND6), pregnant Wistar rats were fed with a diet supplemented with Lf (1 g/kg) or with an isocaloric control diet until offspring euthanasia. At PND7, pups of both sexes were subjected to experimental HI through the occlusion of the right common carotid artery followed by 60 min of hypoxia (8 % oxygen). Lf prevented HI-induced increased levels of DCFH and lipoperoxidation in hippocampus. Furthermore, Lf enhanced antioxidant defenses including SOD, GPx, and GSH, counteracting HI-induced oxidative stress. HI injury altered the activities of enzymes in the mitochondrial respiratory chain and increased the mitochondrial membrane potential. Both effects were counteracted by Lf supplementation. Lactoferrin prevented oxidative stress and to restored mitochondrial function by upregulating Nrf2 and UCP2 expression following experimental HI. Our results show that even a shorter period of Lf delivery to rat pups is able to improve hippocampal response to neonatal hypoxia-ischemia, reversing initial mechanisms of damage in the cascade of HI injury.

Reactive oxygen species (ROS) are chemical moieties that in physiological concentrations serve as fast-acting signaling molecules important for cellular homeostasis. However, their excess either due to overproduction or inability of the antioxidant system to inactivate them results in oxidative stress, contributing to cellular dysfunction and tissue damage. In tendons, which are hypovascular, hypocellular, and composed predominantly of extracellular matrix (ECM), particularly collagen I, ROS likely play a dual role: regulating cellular processes such as inflammation, proliferation, and ECM remodeling under physiological conditions, while contributing to tendinopathy and impaired healing when dysregulated. This review explores the sources of ROS in tendons, including NADPH oxidases and mitochondria, and their role in key processes such as tissue adaptation to mechanical load and injury repair, also in systemic conditions such as diabetes. In addition, we integrate the emerging perspective that calcium signaling-mediated by mechanically activated ion channels-plays a central role in tendon mechanotransduction under daily mechanical loads. We propose that mechanical overuse (overload) may lead to hyperactivation of calcium channels, resulting in chronically elevated intracellular calcium levels that amplify ROS production and oxidative stress. Although direct evidence linking calcium channel hyperactivity, intracellular calcium dysregulation, and ROS generation under overload conditions is currently circumstantial, this review aims to highlight these connections and identify them as critical avenues for future research. By framing ROS within the context of both adaptive and maladaptive responses to mechanical load, this review provides a comprehensive synthesis of redox biology in tendon injury and repair, paving the way for future work, including development of therapeutic strategies targeting ROS and calcium signaling to enhance tendon recovery and resilience.

Mitochondria are key regulators of inflammatory responses and mitochondrial dysfunction is closely linked to various inflammatory diseases. Increasing genetic and experimental evidence suggests that mitochondria play a critical role in inflammatory bowel disease (IBD). In the complex environment of the intestinal tract, intestinal epithelial cells (IECs) and their mitochondria possess unique phenotypic features, shaping each other and regulating intestinal homeostasis and inflammation through diverse mechanisms. Here, we focus on intestinal inflammation in IBD induced by mitochondrial damage-associated molecular patterns (mtDAMPs), which comprise mitochondrial components and metabolic products. The pathogenic mechanisms of mtDAMP signaling pathways mediated by two major mtDAMPs, mitochondrial DNA (mtDNA) and mitochondrial reactive oxygen species (mtROS), are discussed.

Mitochondrial (mt)ROS, insufficient NAD + , and cellular senescence all contribute to the decrease in bone formation with aging. ROS can cause senescence and decrease NAD + , but it remains unknown whether these mechanisms mediate the effects of ROS in vivo . Here, we generated mice lacking the mitochondrial antioxidant enzyme Sod2 in osteoblast lineage cells targeted by Osx1-Cre and showed that Sod2 ΔOsx1 mice had low bone mass. Osteoblastic cells from these mice had impaired mitochondrial respiration and attenuated NAD + levels. Administration of an NAD + precursor improved mitochondrial function in vitro but failed to rescue the low bone mass of Sod2 ΔOsx1 mice. Single-cell RNA-sequencing of bone mesenchymal cells indicated that ROS had no significant effects on markers of senescence but disrupted parathyroid hormone signaling, iron metabolism, and proteostasis. Our data supports the rationale that treatment combinations aimed at decreasing mtROS and senescent cells and increasing NAD + should confer additive effects in delaying age-associated osteoporosis.

Parkinson's disease (PD) is a multi-system disorder characterized histopathologically by degeneration of dopaminergic neurons in the substantia nigra pars compacta. While the etiology of PD remains multifactorial and complex, growing evidence suggests that cellular metabolic dysfunction is a critical driver of neuronal death. Defects in cellular metabolism related to energy production, oxidative stress, metabolic organelle health, and protein homeostasis have been reported in both neurons and immune cells in PD. We propose that these factors act synergistically in immune cells to drive aberrant inflammation in both the CNS and the periphery in PD, contributing to a hostile inflammatory environment which renders certain subsets of neurons vulnerable to degeneration. This review highlights the overlap between established neuronal metabolic deficits in PD with emerging findings in central and peripheral immune cells. By discussing the rapidly expanding literature on immunometabolic dysfunction in PD, we aim to draw attention to potential biomarkers and facilitate future development of immunomodulatory strategies to prevent or delay the progression of PD.

Anesthetics such as ketamine and thiopental, commonly used for inducing unconsciousness, have distinct effects on neuronal activity, metabolism, and cardiovascular and respiratory systems. Ketamine increases heart rate and blood pressure while preserving respiratory function, whereas thiopental decreases both and can cause respiratory depression. This study investigates the impact of ketamine (100 mg/kg) and thiopental (45 mg/kg) on ultraweak photon emission (UPE), oxidative-nitrosative stress, and antioxidant capacity in isolated rat brains. To our knowledge, no previous study has investigated and compared UPE in the presence and absence of anesthesia. Here, we compare the effects of ketamine and thiopental anesthetics with each other and with a non-anesthetized control group. Ketamine increased UPE, lipid peroxidation, and antioxidant enzyme activity while reducing thiol levels. Conversely, thiopental decreased UPE, oxidative markers, and antioxidant enzyme activity, while increasing thiol levels. UPE was negatively correlated with thiol levels and positively correlated with oxidative stress markers. These findings suggest that the contrasting effects of ketamine and thiopental on UPE are linked to their differing impacts on brain oxidative stress and antioxidant capacity. This research suggests a potential method to monitor brain oxidative stress via UPE during anesthesia, and opens up new ways for understanding and managing anesthetic effects.

Growth-Associated Protein-43 (GAP-43) is a calmodulin-binding protein, originally found in neurons, that in skeletal muscle regulates the handling of intracellular Ca2+ dynamics. According to its role in Ca2+ regulation, myotubes from GAP-43 knockout (GAP-43-/-) mice display alterations in spontaneous Ca2+ oscillations and increased Ca2+ release. The emerging hypothesis is that GAP-43 regulates CaM interactions with RyR and DHPR Ca2+ channels. The loss of GAP-43 promotes cardiac hypertrophy in newborn GAP-43-/- mice, extending the physiological role of GAP-43 in cardiac muscle. We investigated the role of GAP-43 in cardiomyocytes derived from the hearts of GAP-43-/- mice, evaluating intracellular Ca2+ variations and the correlation with the levels of reactive oxygen species (ROS), considering their importance in cardiovascular physiology. In GAP-43-/- cardiomyocytes, we found the increased expression of markers of cardiac hypertrophy, Ca2+ alterations, and high mitochondria ROS levels (O2•-) together with increased oxidized functional proteins. Treatment with a CaM inhibitor (W7) restored Ca2+ and ROS alterations, possibly due to high mitochondrial Ca2+ entry by a mitochondrial Ca2+ uniporter. Indeed, Ru360 was able to abolish O2•- mitochondrial production. Our results suggest that GAP-43 has a key role in the regulation of Ca2+ and ROS homeostasis, alterations to which could trigger heart disease.

Epilepsy, affecting approximately 50 million individuals worldwide, is a neurological disorder characterized by recurrent seizures. Mitochondrial dysfunction and oxidative stress are critical factors in its pathophysiology, leading to neuronal hyperexcitability and cell death. Because of the multiple mitochondrial pathways that can be involved in epilepsy and mitochondrial dysfunction, it is optimal to treat epilepsy with multiple antioxidants in combination. Recent advancements highlight the potential of antioxidant therapy as a novel treatment strategy. This approach involves tailoring antioxidant interventions-such as melatonin, idebenone, and plant-derived compounds-based on individual mitochondrial health, including mitochondrial DNA mutations and haplogroups that influence oxidative stress susceptibility and treatment response. By combining antioxidants that target multiple pathways, reducing oxidative stress, modulating neurotransmitter systems, and attenuating neuroinflammation, synergistic effects can be achieved, enhancing therapeutic efficacy beyond that of a single antioxidant on its own. Future directions include conducting clinical trials to evaluate these combination therapies, and to translate preclinical successes into effective clinical interventions. Targeting oxidative stress and mitochondrial dysfunction through combination antioxidant therapy represents a promising adjunctive strategy to modify disease progression and improve outcomes for individuals living with epilepsy.

A liberal supplemental oxygen approach is recommended for all severely injured trauma patients despite limited evidence. Liberal oxygen administration may cause oxidative stress. The aim of this study was to investigate the effect of an early restrictive oxygen strategy versus a liberal oxygen strategy in adult trauma patients on biomarkers of oxidative stress within 48 h of hospital admission.

This was a single-centre, sub-study of an international, randomised controlled trial TRAUMOX2. In TRAUMOX2, patients were randomised shortly after trauma to a restrictive oxygen strategy (arterial oxygen saturation target of 94 %) or a liberal oxygen strategy (12-15 L of oxygen per minute or fraction of inspired oxygen of 0.6-1.0) for 8 h. Blood samplings were performed at four time points within 48 h after randomisation: upon arrival at the trauma centre, and at eight, 24, and 48 h post-randomisation. The primary outcome was the plasma level of malondialdehyde (MDA) 24 h post-randomisation. Secondary outcomes were numerous, and included the level of MDA at other time points, superoxide dismutase (SOD) at all time points, 30-day mortality, and major respiratory complications.

The sub-study included 90 adult trauma patients. The median MDA levels at 24 h post-randomisation was 60.9 μM (95 % CI 49.5 to 73.4) in the restrictive oxygen group and 56.7 μM (95 % CI 46.9 to 68.2) in the liberal oxygen group, corresponding to a difference of -4.2 μM (95 % CI -19.8 to 10.5; P = 0.35). No significant differences were found in MDA or SOD at the other time points either. Neither did we find a significant difference in 30-day mortality or major respiratory complications.

In this sub-study of the TRAUMOX2 trial, no significant differences were found in biomarkers of oxidative stress between a restrictive oxygen strategy and liberal oxygen strategy in adult trauma patients.

Free radicals, including reactive oxygen species (ROS) and reactive nitrogen species (RNS), induce oxidative stress. This stress plays crucial roles in cellular signaling, stress response, and disease progression, making the quantification of free radicals essential for understanding oxidative stress mechanisms. Here, we present a high-throughput fluorescence-based protocol for measuring the presence of total free radicals, including ROS and RNS, in the whole adult Drosophila melanogaster (fruit fly). The protocol involves homogenizing whole adult flies in PBS and treating only the supernatant of the lysate with dichlorodihydrofluorescein-DiOxyQ (DCFH-DiOxyQ), which then converts into a fluorescent molecule, dichlorofluorescein (DCF), upon reacting with free radicals. The level of fluorescence is directly proportional to the amount of free radicals present in the sample. This protocol offers simplicity, scalability, and adaptability, making it ideal for studying oxidative stress in the model organism Drosophila and its different tissues under different dietary regimes, environmental stresses, genetic mutations, or pharmacological treatments. It is to be noted that the protocol uses a kit from Abcam, which has been used to measure free radicals in mice, rats, human blood, and cell lines. It can also be applied to biofluids, culture supernatants, and cell lysates, making it suitable for a wide range of sample types beyond whole organisms or tissues. However, due to our research focus and expertise, here we describe a detailed protocol to measure free radicals responsible for inducing oxidative stress only in fruit flies. Key features • Quantifies total free radicals including ROS and RNS levels in adult Drosophila melanogaster using a fluorescence-based approach for oxidative stress studies. • Suitable for high-throughput analysis with a 96-well black plate format, simultaneously enabling efficient handling of multiple samples and standards. • Adaptable to different experimental conditions, including diverse ROS-inducing treatments and mutations in Drosophila. • Offers detailed instructions for reagent preparation, sample homogenization, fluorescence measurement, normalization, and statistical analysis of data to ensure reproducibility and accuracy across research settings. Graphical overview Schematic workflow of the assay. Whole adult fruit flies are homogenized in PBS buffer and centrifuged. The clear supernatant is carefully transferred into new tubes for further treatment with different reagents, loaded into a clear-bottom black 96-well plate, and treated with another set of reagents. The plate is then incubated, and fluorescence is measured using the Agilent BioTek Synergy H1 plate reader.

Mitochondria play a central role in nutrient metabolism, besides being responsible for the production of adenosine triphosphate (ATP), the main source of cellular energy. However, the ATP production process is associated with the generation of reactive oxygen species (ROS), which excessive accumulation can cause mitochondrial dysfunction. This dysfunction, in turn, causes the accumulation of fatty acids in the adipose tissue, triggering a local inflammatory process that can evolve into systemic inflammation. In women with obesity, an increase in lipid levels in the placental environment is observed. The high presence of fatty acids compromises the structural integrity and mitochondrial membrane, culminating in the release of ROS. This process damages the DNA of placental cells and causes an inflammatory state, affecting metabolic efficiency. This vicious cycle is characterized by defects in mitochondrial ATP production, which can lead to lipid accumulation and inflammation. In pregnant women with obesity, these mitochondrial changes play a determining role in pregnancy outcomes. Hence, the objective of this study was to search the literature to review the impact of mitochondrial dysfunction in the maternal obesity.

Enzymes are widely distributed in organelles of cells, which are capable of carrying out specific catalytic reactions. In general, several enzymes collaborate to facilitate complex reactions and engage in vital biochemical processes within cells, which are also called cascade systems. The cascade systems are highly efficient, and their dysfunction is associated with a multitude of endogenous diseases. The advent of nanotechnology makes it possible to mimic these cascade systems in nature and realize partial functions of natural biological processes both in vitro and in vivo. To emphasize the significance of artificial cascade systems, mimicomes is first proposed, a new concept that refers to the artificial cascade catalytic systems. Typically, mimicomes are able to mimic specific natural biochemical catalytic processes or facilitate the overall catalytic efficiency of cascade systems. Subsequently, the evolution and development of different types of mimicomes in recent decades are elucidated exhaustedly, from the natural enzyme-based mimicomes (immobilized enzyme and vesicle mimicomes) to the nanozyme-based mimicomes and enzyme-nanozyme hybrid mimicomes. In conclusion, the remaining challenges in the design of multifunctional mimicomes and their potential applications are summarized, offering insights into their future prospects.

Manganese superoxide dismutase (MnSOD/SOD2) is an essential mitochondrial enzyme that detoxifies superoxide radicals generated during oxidative respiration. MnSOD/SOD2 lysine 68 acetylation (K68-Ac) is an important post-translational modification (PTM) that regulates enzymatic activity, responding to nutrient status or oxidative stress, and elevated levels have been associated with human illness. To determine the in vivo role of MnSOD-K68 in the heart, we used a whole-body non-acetylation mimic mutant (MnSODK68R) knock-in mouse. These mice exhibited several cardiovascular phenotypes, including lower blood pressure, decreased ejection fraction, and importantly, dilated cardiomyopathy, as evidenced by echocardiography at four months of age. In addition, both mouse embryo fibroblasts (MEFs) and cardiovascular tissue from MnSODK68R/K68R mice exhibited an increase in cellular senescence. Finally, MnSODK68R/K68R mouse hearts also showed an increase in lipid peroxidation. We conclude that constitutively active MnSOD detoxification activity, lacking the normal switch between non-acetylated and acetylated forms, dysregulates mitochondrial physiology during development, leading to dilated cardiomyopathy.

Cellular heterogeneity plays a crucial role in biotechnological processes, significantly influencing metabolic activity, product yield, and process consistency. This review explores the different dimensions of cellular heterogeneity, focusing on its manifestation at both single-cell and population levels. The study examines how factors such as asymmetric cell division, age, and environmental conditions contribute to functional diversity within cell populations, with an emphasis on microorganisms like yeast. Age-related cellular heterogeneity, in particular, is highlighted for its impact on metabolic pathways, mitochondrial function, and secondary metabolite production, which directly affect bioprocess outcomes. Furthermore, the review discusses advanced techniques for detecting and managing heterogeneity, including surface marker-based approaches, which utilize proteins, polysaccharides, and lipids, and label-free methods that leverage cellular volume and physical properties for separation. Understanding and controlling cellular heterogeneity is essential for optimizing industrial bioprocesses, improving yield, and ensuring product quality. The review also underscores the potential of emerging biotechnological tools, such as real-time single-cell analysis and microfluidic devices, in enhancing separation techniques and managing cellular diversity for better process efficiency and robustness.

Cardiac function declines with age, impairing exercise tolerance and negatively impacting healthy aging. However, mechanisms driving age-related declines in cardiac function are not fully understood.

We examined mechanisms underlying age-related cardiac dysfunction using 3- and 24-month-old wild-type mice fed ad libitum or 24-month-old wild-type mice subjected to 70% calorie restriction (CR) starting at 2-month-old. In addition, cardiac aging phenotypes and mitochondrial biogenesis were also analyzed in 25-month-old cardiac-specific Hint1 knockout mice, 24-month-old CAG-Caren Tg mice, and 24-month-old wild-type mice injected with AAV6-Caren.

We observed inactivation of mitochondrial biogenesis in hearts of aged mice. We also showed that activity of the BAF chromatin remodeling complex is repressed by HINT1, whose expression in heart increases with age, leading to decreased transcription of Tfam, which promotes mitochondrial biogenesis. Interestingly, CR not only suppressed age-related declines in cardiac function and mitochondrial biogenesis but blocked concomitant increases in cardiac HINT1 protein levels and maintained Tfam transcription. Furthermore, expression of the lncRNA Caren, which inhibits Hint1 mRNA translation, decreased with age in heart, and CR suppressed this effect. Finally, decreased HINT1 expression due to Caren overexpression antagonized age-related declines in mitochondrial biogenesis, ameliorating age-related cardiac dysfunction, exercise intolerance, and exercise-induced cardiac damage and subsequent death of mice.

Our findings suggest that mitochondrial biogenesis in cardiomyocytes decreases with age and could underlie cardiac dysfunction, and that the Caren-HINT1-mitochondrial biogenesis axis may constitute a mechanism linking CR to resistance to cardiac aging. We also show that ameliorating declines in mitochondrial biogenesis in cardiomyocytes could counteract age-related declines in cardiac function, and that this strategy may improve exercise tolerance and extend so-called "healthy life span".

The essential oil from Lippia origanoides (EOLO) is employed in traditional medicine as it has both antimicrobial and anti-inflammatory properties. The current investigation first evaluated the EOLO's cytotoxic activity in tumour (SiHa and HT-29) and non-tumour (human lymphocyte) cells by MTT. The effect on ROS production was further evaluated in cancer cells by fluorimetry. The oil's mutagenic and antifungal activities were also evaluated using, respectively, the in vitro micronucleus test and the broth microdilution method. The EOLO displayed significant cytotoxicity in both cancer cell lines, with IC50 values of 20.2 μg/mL and 24.3 μg/mL for HT-29 and for SiHa cell lines, respectively. EOLO increased ROS production, was unable to raise the micronucleus frequencies and significantly reduced the cytokinesis block proliferation indices, revealing its anti-proliferative action. The results demonstrate that EOLO is devoid of mutagenic activity but possesses significant activity against tumour and non-tumour human cells, reinforcing its biological potential.

Acetylcholinesterase inhibition, the principal mechanism of acute organophosphorus compound toxicity, cannot explain neuropsychiatric symptoms occurring after exposure to low organophosphate concentrations causing no cholinergic symptoms. Organophosphate-triggered oxidative stress has increasingly come into focus, occurring when the action of reactive oxygen species, generated from free radicals, is not compensated by antioxidant free radical scavengers. Being nucleophilic, organophosphates can easily accept an electron, thereby generating free radicals. Organophosphates inhibit the antioxidant paraoxonase, and reactive oxygen species are produced during organophosphate metabolism. Organophosphates disrupt the function of mitochondria, the principal source of free radicals. Organophosphates also induce neuroinflammation, which generates reactive oxygen species, and reactive oxygen species in turn stimulate neuroinflammation. Markers of reactive oxygen species are elevated in vitro and in vivo after exposure to organophosphates and in individuals professionally exposed to organophosphates. This most probably contributes to the pathogenesis of the intermediate syndrome, chronic organophosphate-induced neuropsychiatric disorders and neurodegeneration occurring in patients after organophosphate exposure. Evidence for beneficial effects of antioxidants in organophosphate poisoning is discussed.

Reactive oxygen species (ROS) are redox-signaling molecules involved in aging and lifestyle-related diseases. In the brain, in addition to the production of ROS as byproducts of metabolism, expression of ROS synthases has recently been demonstrated, suggesting possible involvement of ROS in various brain functions. This review highlights current knowledge on the relationship between ROS and brain functions, including their contribution to age-related decline in synaptic plasticity and cognitive function. While most studies demonstrate either the positive or negative effects of ROS on synaptic plasticity, the dual effects of ROS at individual synapses have been demonstrated recently in the mouse cerebellum. Furthermore, the cooperative interaction between these two effects determines the direction of synaptic plasticity. It is anticipated that further elucidation of both the positive and negative effects of ROS on brain function will lead to the development of more effective therapeutic strategies with fewer side effects for ROS-related brain dysfunction.

Neuropathic pain, a debilitating condition arising from somatosensory system damage, significantly impacts quality of life, leading to anxiety, self-mutilation, and depression. Oxidative and nitrosative stress, an imbalance between reactive oxygen and nitrogen species (ROS/RNS) and antioxidant defenses, plays a crucial role in its pathophysiology. While reactive species are essential for physiological functions, excessive levels can cause cellular component damage, leading to neuronal dysfunction and pain. This review highlights the complex interactions between reactive species, antioxidant systems, cell signaling, and neuropathic pain. We discuss the physiological roles of ROS/RNS and the detrimental effects of oxidative and nitrosative stress. Furthermore, we explore the potential of manganese porphyrins, compounds with antioxidant properties, as promising therapeutic agents to mitigate oxidative stress and alleviate neuropathic pain by targeting key cellular pathways involved in pain. Further research is needed to fully understand their therapeutic potential in managing neuropathic pain in human and non-human animals.

One of the major causes of senescence is oxidative stress caused by ROS, which is mainly generated from dysfunctional mitochondria. Strategies to limit mitochondrial ROS production are considered important for reversing senescence, but effective approaches to reduce them have not yet been developed. In this study, we screened the secondary metabolites that plants produce under oxidative stress and discovered sauchinone as a potential candidate. Sauchinone induced mitochondrial function recovery, enabling efficient electron transport within the electron transport chain (ETC). This led to a decrease in ROS production, a byproduct of inefficient electron transport. The reduction in ROS by sauchinone rejuvenated senescence-associated phenotypes. To understand the underlying mechanism by which sauchinone rejuvenates senescence, we carried out RNA sequencing and found VAMP8 as a key gene. VAMP8 was downregulated by sauchinone. Knockdown of VAMP8 decreased mitochondrial ROS levels and subsequently rejuvenated mitochondrial function, which was similar to the effect of sauchinone. Taken together, these studies revealed a novel mechanism by which sauchinone reduces mitochondrial ROS production by regulating mitochondrial function and VAMP8 expression. Our results open a new avenue for aging research to control senescence by regulating mitochondrial ROS production.

Reactive oxygen species (ROS) contribute to aging by mainly damaging cellular organelles and DNA. Although strategies to reduce ROS production have been proposed as important components of anti-aging therapy, effective mechanisms to lower ROS levels have not yet been identified. Here, we screened natural compounds frequently used as cosmetic ingredients to find substances that reduce ROS levels. Magnolia officinalis (M. officinalis) extract significantly lowered the levels of ROS in senescent fibroblasts. A novel mechanism by which M. officinalis extract restores mitochondrial function to reduce ROS, a byproduct of inefficient electron transport, was discovered. The reduction of ROS by M. officinalis extracts reversed senescence-associated phenotypes and skin aging. Then, honokiol was demonstrated as a core ingredient of M. officinalis extract that exhibits antioxidant effects. Honokiol functions as an oxygen radical scavenger through redox processes, also significantly reduced ROS levels by restoring mitochondrial function. In summary, our study identified a novel mechanism by which M. officinalis extract reverses aging and skin aging by reducing ROS through restoring mitochondrial function. These new findings will not only expand our understanding of aging and associated diseases, but also provide new approaches to anti-aging treatments.

Drug resistance is a major barrier to cancer therapies and remains poorly understood. Recently, non-mutational mechanisms of drug resistance have been proposed where a more plastic metabolic response can play a major role. Here, we show that upon drug resistance, glioblastoma (GBM) cells have increased oxidative stress, mitochondria function, and protein aggregation. Gamma (γ)-glutamylcyclotranserase (GGCT), an enzyme in the γ-glutamyl cycle for glutathione production, located on chromosome 7 which is commonly amplified in GBM is also increased upon resistance. We further observe that the byproduct of GGCT-pyroglutamic acid-can bind aggregating proteins and that genetic and pharmacological inhibition of GGCT prevents protein aggregation. Finally, we found increased protein aggregation, GGCT expression, and pyroglutamic acid staining in recurrent GBM patient samples, adjacent non-tumor brain, and Alzheimer's brains. These findings suggest a new pathway for protein aggregation within drug resistant brain cancer that should be further studied in other brain disorders.

The KIT receptor is a transmembrane protein found on the surface of many different cell types. Mutant forms of KIT are drivers of myeloid neoplasms, including systemic mastocytosis. The KIT D816V mutation is the most common, leading to constitutive activation of the receptor and its downstream targets, and it is highly resistant to c-KIT inhibitors. Metabolic rewiring is a common trait in cancer. We analyzed the metabolic profile induced by the KIT D816 mutation, measuring mitochondrial parameters in two myeloid cell lines. We found that the KIT D816V mutation causes a significant increase in mitochondrial abundance and activity associated with superoxide production, which could promote DNA instability. Functional and morphologic changes in mitochondria were associated with reduced levels of BNIP3 protein expression. We also detected low BNIP3 levels in clinical acute myeloid leukemia samples harboring D816V mutations. In addition, we have found constitutive mTOR activation in mutated cells, a pathway that has been shown to regulate autophagy. Our data suggest that KIT D816V increases mitochondrial activity through downregulation of BNIP3 expression, which increases mitochondrial number through the autophagy pathway. Alterations in the cellular metabolism induced by the KIT D816V mutation could be therapeutically exploited.

Type 1 diabetes (T1D) is a multifactorial disease involving genetic and environmental factors, including viral infection. We investigated the impact of interferon alpha (IFN-α), a cytokine produced during the immune response to viral infection or the presence of un-edited endogenous double-stranded RNAs, on human β-cell physiology. Intravital microscopy on transplanted human islets using a β-cell-selective reactive oxygen species (ROS) biosensor (RIP1-GRX1-roGFP2), revealed a subset of human β-cells that acutely produce ROS in response to IFN-α. Comparison to Integrated Islet Distribution Program (IIDP) phenotypic data revealed that healthier donors had more ROS accumulating cells. I n vitro IFN-α treatment of human islets similarly elicited a heterogenous increase in superoxide production that originated in the mitochondria. To determine the unique molecular signature predisposing cells to IFN-α stimulated ROS production, we flow sorted human islets treated with IFN-α. RNA sequencing identified genes involved in inflammatory and immune response in the ROS-producing cells. Comparison with single cell RNA-Seq datasets available through the Human Pancreas Analysis Program (HPAP) showed that genes upregulated in ROS-producing cells are enriched in control β-cells rather than T1D donors. Combined, these data suggest that IFN-α stimulates mitochondrial ROS production in healthy human β-cells, potentially predicting a more efficient antiviral response.

Selective imaging of superoxide anion is important for understanding its role in cell membrane biology, but is often a challenging task because of the lack of an effective fluorescence probe. In this study, a new near-infrared fluorescent probe (SHX-O) that can target cell membrane was developed for imaging superoxide anion. SHX-O was designed by simultaneously incorporating a sulfonated bis-indole and a diphenylphosphinyl recognition group into the hemicyanine moiety. The probe itself showed a rather weak fluorescence due to the hemicyanine's hydroxyl substitution; however, its reaction with superoxide anion caused a large enhancement of near-infrared fluorescence at 790 nm. Moreover, SHX-O exhibited not only high selectivity for superoxide anion over other reactive oxygen species, but also specific cell membrane localization, which may be attributed to the probe's amphiphilic structure. Using the probe, fluorescence imaging of cell membrane superoxide anion produced in the presence of xanthine oxidase and xanthine has been achieved in living cells. We believe that SHX-O may serve as a potential tool for imaging and investigating superoxide anion of cell membrane.

Reactive oxygen species (ROS) have been implicated in behaviors induced by acute or repeated cocaine or methamphetamine administration in rodents. In the present study, we investigated the involvement of ROS in behavioral changes induced by nicotine administration and dopamine (DA) transmission changes in the nucleus accumbens (NAc) of rats. Rats were given repeated saline or nicotine (0.4 mg/kg) administration once daily for seven days, and the induction of hyperlocomotor activity, and oxidative stress marker expression induced by the increase in ROS production in the NAc were measured on day 7. We also tested the effect of ROS scavengers on repeated nicotine-induced hyperlocomotor activity and nicotine self-administration, and DA levels in the NAc. Repeated nicotine administration induced hyperlocomotor activity and decreased the expression of oxidative stress markers, such as superoxide dismutase-1 and glutathione peroxidase 1/2, by elevating ROS production in the NAc. Pretreatment with the nonspecific ROS scavenger PBN and the superoxide-selective scavenger TEMPOL significantly attenuated nicotine-induced hyperlocomotor activity without impairing motor function in nicotine-naïve rats on day 7. In addition, in intravenous nicotine self-administration study, TEMPOL significantly reduced nicotine-taking behavior without affecting food intake in nicotine-naïve rats. Furthermore, TEMPOL pretreatment prevented nicotine effects on stimulated DA release in the NAc, which was associated with nicotine-induced behavioral changes. Taken together, these findings suggest that increased ROS production in the NAc contributes to the neuropharmacological properties of nicotine.

Glioblastoma (GBM) is a prevalent and refractory type of brain tumor. Over the past two decades, there have been minimal advancements in GBM therapy. The current standard treatment involves surgical excision followed by radiation and chemotherapy. Compared to other tumors, GBM is more challenging to treat due to the presence of glioma stem-like cells (GSCs) and the blood-brain barrier, resulting in an extremely low survival rate. Mitochondria play a critical role in tumor respiration, metabolism, and multiple signaling pathways involved in tumor formation, progression, and cell apoptosis. Consequently, mitochondria represent promising targets for developing novel anticancer agents, including those targeting oxidative phosphorylation, reactive oxygen species (ROS), mitochondrial transfer, and mitophagy. This review outlines the mitochondrial-related therapeutic targets in GBM, highlighting the potential of mitochondria as a target for GBM treatment.

Myocardial ischemia/reperfusion (I/R) injury has high morbidity and mortality rates, posing a significant burden on society. There is an urgent need to understand its pathogenesis and develop effective treatments. Reactive oxygen species (ROS) are crucial for the development of myocardial I/R injury, and inhibiting ROS overproduction is one of the most critical ways to delay myocardial I/R injury. Sirtuins are a group of nicotinic adenine dinucleotide ( +)-dependent histone deacetylases whose members can regulate ROS by modulating various biological processes. Numerous studies have shown that Sirtuins play an essential role in the progression of myocardial I/R injury by regulating ROS. This study focuses on the relationship between myocardial I/R injury and ROS, Sirtuins and ROS, discusses the role of Sirtuins in regulating ROS in myocardial I/R, and summarizes the therapeutic modalities aimed at targeting Sirtuins to modulate ROS in myocardial I/R injury, thereby guiding future research endeavors.

Noise-induced hearing loss (NIHL) constitutes a significant global health issue for which there is no effective treatment. The loss of cochlear hair cells and associated synaptopathy are common causes of hearing impairment. One primary mechanism implicated in NIHL is the accumulation of reactive oxygen species (ROS), which ultimately overwhelms cochlear cells. ROS are detected in the cochlea immediately after noise exposure and persist for at least a week. Within cells, ROS are primarily generated in mitochondria as byproducts of cellular metabolism. Elamipretide is a synthetic tetrapeptide known to concentrate in mitochondria, improving mitochondrial function and reducing ROS production. To test the hypothesis that elamipretide treatment mitigates NIHL, 16-week-old male and female CBA/J mice were exposed to 8-16 kHz octave-band noise (OBN) at 98 dB SPL for 2 hours. Elamipretide was administered intraperitoneally immediately after noise exposure and continued for 2 weeks. Efficacy was evaluated based on auditory brainstem response (ABR) thresholds, wave amplitudes, and wave latencies in treated and control groups. Results showed that OBN-exposed mice exhibited an elevation in ABR thresholds at 16 and 32 kHz and a reduction in ABR wave-I amplitude at 32 kHz, although wave-I latencies were not affected at 16 or 32 kHz. Elamipretide treatment prevented the OBN-induced elevation of ABR thresholds and the attenuation of wave-I amplitude. These findings provide proof of concept that mitochondrial-targeted elamipretide can prevent NIHL in a mammalian model and highlight its potential to protect against NIHL in humans.

TGF-β-activated kinase 1 binding protein 2 (TAB2) is an intermediary protein that links Tumor necrosis factor receptor 1 (TNFR1) and other receptor signals to the TGF-β-activated kinase 1 (TAK1) signaling complex. TAB2 frameshift mutations have been linked to dilated cardiomyopathy (DCM), while the exact mechanism needs further investigation.

In this study, we generated a TAB2 compound heterozygous knockout cell line in induced pluripotent stem cells (iPSCs) derived from a healthy individual using CRISPR/Cas9 technology. IPSCs are not species-dependent, are readily accessible, and raise fewer ethical concerns.

TAB2 disruption had no impact on the cardiac differentiation of iPSCs and led to confirmed TAB2 deficiency in human iPSC-derived cardiomyocytes (hiPSC-CMs). TAB2-deficient hiPSC-CMs were found to develop phenotypic features of DCM, such as distorted sarcomeric ultrastructure, decreased contractility and energy production, and mitochondrial damage at day 30 post differentiation. Paradoxically, TAB2 knockout cell lines showed abnormal calcium handling after 40 days, later than reduced contractility, suggesting that the main cause of impaired contractility was abnormal energy production due to mitochondrial damage. As early as day 25, TAB2 knockout cardiomyocytes showed significant mitochondrial calcium overload, which can lead to mitochondrial damage. Furthermore, TAB2 knockout activated receptor-interacting protein kinase 1 (RIPK1), leading to an increase in mitochondrial calcium uniporter (MCU) expression, thereby augmenting the uptake of mitochondrial calcium ions. Finally, the application of the RIPK1 inhibitor Nec-1s prevents the progression of these phenotypes.

In summary, TAB2 abatement cardiomyocytes mimic dilated cardiomyopathy in vitro. This finding emphasizes the importance of using a human model to study the underlying mechanisms of this specific disease. More importantly, the discovery of a unique pathogenic pathway introduces a new notion for the future management of dilated cardiomyopathy.

Breathing hyperoxic gas is common in diving and accelerates fatigue after prolonged and repeated exposure. The mechanism(s) remain unknown but may be related to increased oxidants that interfere with skeletal muscle calcium trafficking or impaired aerobic ATP production. To determine these possibilities, C57BL/6J mice were exposed to hyperbaric oxygen (HBO2) for 4 h on three consecutive days or remained in room air. Postfinal exposure, fatigue was determined by grip strength and run-to-exhaustion tests. Other measurements included indices of oxidant stress and antioxidant defenses, mitochondrial bioenergetics, caffeine-induced sarcoplasmic reticulum-calcium release, and S-nitrosylation of ryanodine receptor 1 (RyR1). Despite grip strength being unaffected by repeated HBO2 exposure, mean running time was reduced by 50%. In skeletal muscle from HBO2 exposed mice, superoxide production was significantly increased, resulting in elevated lipid and DNA (nuclear and mitochondrial) oxidation. Accompanying increased oxidant stress was a reduction in glutathione content and increased Sod1 and Hmox1 gene expression; Ucp3 mRNA was reduced. Mitochondrial respiration, mitochondrial membrane potential, and NAD+/NADH were not influenced by HBO2. In contrast, caffeine-induced sarcoplasmic reticulum (SR)-calcium release was reduced by 66% and S-nitrosylation of RyR1 was increased by 45%. Exposing mice to repeated HBO2 increases oxidant stress that activates some antioxidant defenses. Mitochondrial function is not altered and could be related to decreased production of UCP3 that serves to maintain the electrochemical proton gradient. S-nitrosylation of RyR1 may promote SR-calcium leak and reduce content, a potential mechanism for repeated HBO2-induced fatigue.NEW & NOTEWORTHY Breathing hyperoxic gas during prolonged and repeated dives causes fatigue but the mechanisms are unknown. Here, we show in mice exposed to repeated hyperbaric oxygen that running fatigue is accelerated and accompanied by increased skeletal muscle oxidant stress and reduced caffeine-induced sarcoplasmic reticulum (SR)-calcium release. The latter may be due to increased S-nitrosylation of ryanodine receptor 1 (RyR1) and be a mechanism for impaired physical performance after repeated oxygen diving.

Tumor microenvironment (TME) is composed of diverse cell types whose interactions, both direct and indirect, significantly influence tumorigenesis and therapeutic outcomes. Within TME, reactive oxygen species (ROS) are produced by various cells and exhibit a dual role: moderate ROS levels promote tumor initiation and progression, whereas excessive levels induce cancer cell death, influencing the efficacy of anticancer therapies. Inflammasomes, cytosolic multiprotein complexes, are pivotal in multiple stages of tumorigenesis and play a crucial role in establishing the inflammatory TME. By releasing cytokines such as IL-1β and IL-18, inflammasomes contribute to immune cell recruitment and sustain a chronic inflammatory state that supports tumor growth. ROS are critical regulators of inflammasome activation, with the impact of ROS-mediated activation differing across cell types, leading to distinct influences on tumor progression and therapeutic responses. This review explores how ROS drive inflammasome activation in various TME-associated cells and the reciprocal ROS generation induced by inflammasomes, examining their multifaceted impact on tumorigenesis and therapeutic efficacy. By elucidating the complex interplay between ROS and inflammasomes in TME, we provide insights into potential therapeutic approaches that could modulate cancer progression and enhance treatment outcomes.

Organelles such as mitochondria, lysosomes, peroxisomes, and the endoplasmic reticulum form highly dynamic cellular networks and exchange information through sites of physical contact. While each organelle performs unique functions, this inter-organelle crosstalk helps maintain cell homeostasis. Age-related macular degeneration (AMD) is a devastating blinding disease strongly associated with mitochondrial dysfunction, oxidative stress, and decreased clearance of cellular debris in the retinal pigment epithelium (RPE). However, how these occur, and how they relate to organelle function both with the RPE and potentially the photoreceptors are fundamental, unresolved questions in AMD biology. Here, we report the discussions of the "Mitochondria, Lysosomes, and other Organelle Interactions" task group of the 2024 Ryan Initiative for Macular Research (RIMR). Our group focused on understanding the interplay between cellular organelles in maintaining homeostasis in the RPE and photoreceptors, how this could be derailed to promote AMD, and identifying where these pathways could potentially be targeted therapeutically.

Normal brain functioning relies on high aerobic energy production provided by mitochondria. Failure to supply a sufficient amount of energy, seen in different brain disorders, including autism spectrum disorder (ASD), may have a significant negative impact on brain development and support of different brain functions. Mitochondrial dysfunction, manifested in the abnormal activities of the electron transport chain and impaired energy metabolism, greatly contributes to ASD. The aberrant functioning of this organelle is of such high importance that ASD has been proposed as a mitochondrial disease. It should be noted that aerobic energy production is not the only function of the mitochondria. In particular, these organelles are involved in the regulation of Ca2+ homeostasis, different mechanisms of programmed cell death, autophagy, and reactive oxygen and nitrogen species (ROS and RNS) production. Several syndromes originated from mitochondria-related mutations display ASD phenotype. Abnormalities in Ca2+ handling and ATP production in the brain mitochondria affect synaptic transmission, plasticity, and synaptic development, contributing to ASD. ROS and Ca2+ regulate the activity of the mitochondrial permeability transition pore (mPTP). The prolonged opening of this pore affects the redox state of the mitochondria, impairs oxidative phosphorylation, and activates apoptosis, ultimately leading to cell death. A dysregulation between the enhanced mitochondria-related processes of apoptosis and the inhibited autophagy leads to the accumulation of toxic products in the brains of individuals with ASD. Although many mitochondria-related mechanisms still have to be investigated, and whether they are the cause or consequence of this disorder is still unknown, the accumulating data show that the breakdown of any of the mitochondrial functions may contribute to abnormal brain development leading to ASD. In this review, we discuss the multifaceted role of mitochondria in ASD from the various aspects of neuroscience.