Search Results (1,052)

As the direct energy source in organisms, accurate and simple detection of adenosine triphosphate (ATP) is of great significance. Herein, a colorimetric aptasensor for ATP determination was designed by integrating the CRISPR/Cas12a system with an aptamer, and with Prussian blue nanocube and gold nanoparticle co-functionalized MoS₂ (MoS₂-PBNCs-AuNPs) nanozymes. As expected, the introduced CRISPR/Cas12a system and aptamer could efficiently amplify the detection signal and improve the specific recognition ability, respectively. Meanwhile, the catalytic activity of the MoS₂-PBNCs-AuNPs nanozymes can be regulated with the concentration of ATP. The high-affinity binding of ATP to the aptamer competitively inhibited aptamer-crRNA hybridization, causing fewer Cas12 proteins to be activated. As a result, the uncleaved single-stranded DNA (ssDNA) adsorbed onto the surface of nanozymes to effectively enhance their catalytic oxidation capability toward 3,3′,5,5′-tetramethylbenzidine (TMB). According to this phenomenon, this CRISPR-enhanced colorimetric aptasensor can detect down to 0.14 μM ATP with high selectivity, reproducibility, and stability. In addition, acceptable recoveries and low relative standard deviations of the aptasensor for ATP determination suggest that it is promising for application in early detection of clinical-related diseases. Full article

Exposure to hypoxia at high altitudes is significantly associated with impairments in learning and memory functions, as well as abnormalities in neuronal function and synaptic plasticity. Recent research has indicated that mitochondrial reactive oxygen species (mtROS) play a role in regulating microglial activation and mediating neurotoxic damage in the hippocampal CA1 region. Nicotinamide riboside (NR), upon absorption, is rapidly converted into nicotinamide adenine dinucleotide (NAD+), which is involved in the production of mitochondrial adenosine triphosphate (ATP). The potential of NR to protect dendritic spine plasticity in hippocampal CA1 neurons following hypoxia exposure, potentially through the inhibition of microglial activation, warrants further investigation. To this end, a mouse model simulating hypoxia at an altitude of 6000 m over a two-week period, along with a BV2 cells and conditional co-culture of BV2 cells and HT22 cells 1%O₂ hypoxia model, was developed. Behavioral assessments indicated that, relative to the normoxia group, mice subjected to hypoxia exhibited a significant reduction in the time spent in the target quadrant, the distance traveled within the target quadrant, the number of platform crossings, and the novel object recognition index. Furthermore, Golgi staining revealed a marked decrease in the density of dendritic spines in the hippocampal CA1 region in the hypoxia-exposed mice compared to the normoxia group. Subsequently, A daily dosage of 400 mg/kg of NR was administered for two weeks and 0.5 mM NR was used in a conditional co-culture model. Results demonstrated that, in comparison to the hypoxia group, the group receiving combined hypoxia and NR treatment showed significant improvements in the time spent in the target quadrant, the distance traveled within the target quadrant, the number of platform crossings, the novel object recognition index, and the density of dendritic spines in the hippocampal CA1 region. Additionally, transmission electron microscopy indicated a significant increase in the synaptic density of hippocampal neurons in the combined hypoxia exposure and NR treatment group compared to the hypoxia exposure group. Simultaneously, when compared to the hypoxia group, the combination of hypoxia and NR treatment resulted in an increased concentration of mitochondrial ATP. This treatment also partially restored mitochondrial membrane integrity, reduced mtROS levels, decreased the percent of Iba1⁺CD68⁺Iba1⁺ microglia, and lowered the interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNFα), and inducible nitric oxide synthase (iNOS) mRNA levels. These findings indicate that NR treatment may mitigate neurotoxic damage in the hippocampal CA1 region induced by hypoxia exposure, primarily through the attenuation of microglial activation and the reduction in mtROS production. Full article

The increasing utilization of single-use plastics in the food sector poses serious environmental challenges. A circular economy approach, i.e., reusing packaging before recycling, offers a promising solution but raises concerns about cross-contamination between food products. This study investigates how repeated use and cleaning affect the surface topography of plastic food packaging and, in turn, how these changes influence cleaning efficiency and assessment. Recycled polyethylene terephthalate (rPET) trays were subjected to 20 industrial wash cycles with and without detergent concentration of 0.3% v/v at the following temperatures: 55 °C wash, 70 °C rinse. Surface roughness was measured using mechanical and optical techniques. Additionally, trays were roughened with sandpaper of varying grit sizes to simulate mechanical wear during consumer use. Cleanability was assessed using UV fluorescence imaging and adenosine triphosphate (ATP) assays. Results showed no significant increase in surface roughness after 20 wash cycles. However, artificially roughened surfaces retained more food residue, complicating cleaning. The application of UV fluorescence imaging proved more effective than ATP assays in detecting food residues on textured surfaces. These findings support the use of advanced imaging for evaluating the hygiene of reusable packaging and highlight key considerations for implementing circular reuse systems in food packaging. Full article

This study comprehensively evaluated the antimicrobial performance of copper ions against three bacterial species relevant to water systems: E. coli (ATCC 25922), P. aeruginosa (ATCC 27853), and S. epidermidis (ATCC 12228). Disinfection kinetics were determined at three copper concentrations (0.5, 1.5, and 3.3 mg/L) using the Gard model. E. coli exhibited the highest susceptibility, with inactivation rate constants of 0.63, 3.27, and 9.83, achieving complete inactivation at 3.3 mg/L. P. aeruginosa was the most resistant, showing values below 1.0 across all concentrations, while S. epidermidis displayed intermediate responses. Selected experiments further examined the influence of growth phase, temperature, and water chemistry. Exponential-phase cells were more sensitive than stationary-phase cultures, and higher temperatures (37 °C vs. 5 °C) significantly enhanced inactivation. Moderate bicarbonate (50 mg/L) improved bacterial removal by stabilizing soluble Cu²⁺ ions (2.60 lg reduction), whereas elevated calcium and magnesium (Ca²⁺ 100 mg/L, Mg²⁺ 50 mg/L) reduced effectiveness (≤2.10 lg reduction) through competitive interactions. In addition to culture-based methods, adenosine triphosphate (ATP) bioluminescence assays and flow cytometry (FCM) provided complementary insights, confirming early metabolic disruption and membrane damage prior to culturability loss in selected experiments. Full article

In malignant-type gastric cancer, peritoneal dissemination is the most frequent metastatic process and is an inoperable condition for which effective treatment is lacking. Our research has revealed that galectin-4 plays an important role in the peritoneal metastasis of gastric cancer cells. Based on this, we hypothesized that inhibiting galectin-4 could suppress peritoneal metastasis. The inhibitory activity towards galectin-4 binding was evaluated using an enzyme-linked immunosorbent assay, while the suppressive effect on gastric cancer cell proliferation was assessed using an adenosine triphosphate-based cell viability assay. Direct binding to galectin-4 was examined by surface plasmon resonance analysis. Chemically synthesized fucoidan analogs exhibited significant suppressive activity against the proliferation of gastric cancer cells, partly via a galectin-4-mediated pathway. Among the 13 fucoidan analogs tested, analog 10, whose sugar chains composed of repeating 2,3-O-sulfated α(1,4)-linked L-fucose, showed significant inhibitory activity against galectin-4 binding and cell proliferation. 14, the cholestanol-conjugated analog 10, exhibited a pronounced increase in inhibitory activity, consistent with potential multimerization. Molecular docking and site-directed mutagenesis studies revealed that Arginine-45 in galectin-4 is important for binding to fucoidan analogs. In conclusion, fucoidan analogs with a strong affinity for galectin-4 are promising candidates for inhibiting the peritoneal metastasis of galectin-4-positive gastric cancer cells. Full article

Adenosine triphosphate (ATP) is readily released into the extracellular space as an autocrine and paracrine purinergic signaling molecule. We originally reported a genetically encoded, fluorescent protein-based Förster Resonance Energy Transfer (FRET) biosensor that can detect micromolar levels of extracellular ATP. Through mutagenesis of the ATP binding site and optimization of cell-surface display, here we report the development of a second-generation biosensor called ECATS2 with greater than three-fold higher affinity for extracellular ATP. We found that the tether length between the FRET biosensor and the cell surface anchor is critical to optimization of its performance. Furthermore, we demonstrate that the improved sensor can detect extracellular ATP release upon hypoosmotic stress in cultured astrocytes. This new sensor contributes an improved tool for the ratiometric detection of extracellular ATP dynamics and purinergic signaling. Full article

Although orthodontic appliances are widely used in daily practice, they also have their downsides due to the large amount of metal ions released from their surface. In this study, the influence of such released metal ions on the yeast Saccharomyces cerevisiae W303 as a model organism was investigated. Experimental yeast media in which metal ions (iron, aluminum, nickel, chromium, copper, and manganese) were eluted for 3, 7, 14, and 28 days were prepared and then used for yeast cultivation (up to the early stationary growth phase). The growth, increase, and viability of the cells were tested. The mitochondria were isolated from the spheroplasts, and the mitochondrial proteins were obtained and analyzed by liquid chromatography/mass spectrometry. Fortythree significantly altered proteins were identified. They showed significantly reduced expression in all metal-treated groups compared to the control. The metabolic processes for energy supply (glycolysis, gluconeogenesis, tricarboxylic acid cycle, and adenosine triphosphate synthesis) dominated with 50% of the total amount of significantly altered proteins in all samples, but their proportions changed at different time points. The downregulation of mitochondrial proteins such as Atp1, Atp2, and Pet9 under conditions of metal overload suggests a broader impairment of mitochondrial function. Three levels of response to stress can be observed—at relatively low metal ion concentrations in the medium (3 days of elution, approx. 3 mg/L), at medium concentrations (7 days of elution, approx. 5.5 mg/L), and at high concentrations (>8 mg/L, 14 and 28 days of elution), each affecting a specific group of proteins. The results show that mixtures of metal ions in experimental media led to a specific response (in terms of the amount and type of proteins) in each sample type to combat the provoked stress. Full article

During the pathological process of spinal cord injury (SCI), ferroptosis is closely related to mitochondrial homeostasis. Following the occurrence of SCI, the interruption of local blood supply leads to mitochondrial damage within cells and a reduction in Adenosine triphosphate (ATP) production. This results in the loss of transmembrane ion gradients, causing an influx of Ca²⁺ into the cells, which in turn generates a significant amount of Reactive oxygen species (ROS) and reactive nitrogen species. This leads to severe mitochondrial dysfunction and an imbalance in mitochondrial homeostasis. Ferroptosis is a form of programmed cell death that differs from other types of apoptosis, as it is dependent on the accumulation of iron and lipid peroxides, along with their byproducts. The double bond structures in intracellular polyunsaturated fatty acids (PUFA) are particularly susceptible to attack by ROS, leading to the formation of lipid alkyl free radicals. This accumulation of lipid peroxides within the cells triggers ferroptosis. After SCI, the triggering of ferroptosis is closely associated with the “death triangle”—a core network that catalyzes cell death through the interaction of three factors: local iron overload, collapse of antioxidant defenses, and dysregulation of PUFA metabolism (where PUFA are susceptible to attack by reactive ROS leading to lipid peroxidation). These three elements interact to form a central network driving cell death. In the pathological cascade of SCI, mitochondria serve as both a major source of ROS and a primary target of their attack, playing a crucial role in the initiation and execution of cellular ferroptosis. Mitochondrial homeostasis imbalance is not only a key inducer of the “death triangle” (such as the intensification of lipid peroxidation by mitochondrial ROS), but is also reverse-regulated by the “death triangle” (such as the destruction of mitochondrial structure by lipid peroxidation products). Through the cascade reaction of this triangular network, mitochondrial homeostasis imbalance and the “death triangle” jointly drive the progression of secondary damage. This study aims to synthesize the mechanisms by which various therapeutic approaches mitigate SCI through targeted regulation of mitochondrial homeostasis and inhibition of ferroptosis. Unlike previous research, we integrate the bidirectional regulatory relationship between “mitochondrial homeostasis disruption” and “ferroptosis” in SCI, and emphasize their importance as a synergistic therapeutic target. We not only elaborate in detail how mitochondrial homeostasis—including biogenesis, dynamics, and mitophagy—modulates the initiation and execution of ferroptosis, but also summarize recent strategies that simultaneously target both processes to achieve neuroprotection and functional recovery. Furthermore, this review highlights the translational potential of various treatments in blocking the pathological cascade driven by oxidative stress and lipid peroxidation. These insights provide a novel theoretical framework and propose combinatory therapeutic approaches, thereby laying the groundwork for designing precise and effective comprehensive treatment strategies for SCI in clinical settings. Full article

Ametryn (AMT) and clomazone (CLZ) are commonly used herbicides frequently detected in food and water, raising concerns about potential health risks. This study investigated whether AMT and CLZ impair mitochondrial bioenergetics, a key mechanism linked to hepatotoxicity. Mitochondria were isolated from rat liver and incubated with AMT or CLZ (50–200 µM) to assess respiration, membrane potential (Δψ), ATP production, and the activities of respiratory chain complexes and ATP synthase. Both herbicides significantly inhibited state 3 (ADP-stimulated) respiration with glutamate plus malate, without altering state 4 (basal) respiration. Concentrations above 100 µM reduced Δψ and ATP synthesis in glutamate plus malate or succinate-energized mitochondria. Enzymatic assays revealed inhibition of complex I by both herbicides, complex II by CLZ, and ATP synthase by both. These results highlight mitochondrial oxidative phosphorylation disruption by AMT and CLZ; however, further in situ and in vivo studies are necessary to fully understand their hepatotoxic potential. Full article

Background: The ketogenic diet (KD), a high-fat and low-carbohydrate dietary approach, has been used therapeutically in drug-resistant epilepsy and other neurological and metabolic disorders. Recent interest has shifted toward understanding its broader metabolic effects through metabolomics. This review aims to summarize current knowledge on the biochemical mechanisms and therapeutic implications of the KD, with a particular focus on metabolomic profiling and neurological health. Methods: This narrative review synthesizes findings from the last five years of metabolomic studies investigating the biochemical consequences of the KD and its variants, including the classical KD, modified Atkins diet (MAD), medium-chain triglyceride diet (MCT), and low glycemic index treatment (LGIT). The review integrates data on analytical techniques, such as liquid chromatography–mass spectrometry (LC-MS) and gas chromatography–mass spectrometry (GC-MS), and evaluates alterations in key metabolic pathways. Results: The KD significantly modulates energy metabolism, shifting adenosine triphosphate (ATP) production from glycolysis to fatty acid oxidation and ketone body utilization. It affects mitochondrial function, one-carbon metabolism, redox balance, neurotransmitter regulation, and gut–brain axis signaling. Metabolomic profiling has identified β-hydroxybutyrate (βHB) as a key regulatory metabolite influencing mitochondrial respiration. Long-term KD use may impact renal and hepatic function, necessitating clinical caution and individualized nutritional monitoring. Conclusions: Metabolomic analysis provides critical insights into the multifaceted effects of the KD, supporting its role as a targeted metabolic therapy in neurological diseases. However, potential risks linked to prolonged ketosis warrant further investigation. Future studies should focus on personalized applications and long-term safety profiles of KD variants across patient populations. Full article

Background and Objective: With an increase in the number of older citizens in most Western countries, cognitive decline is becoming an increasingly significant issue. Numerous age-related metabolic and physiological changes, such as increased inflammation and oxidative stress, decreased adenosine triphosphate (ATP) production, poorer cardiovascular function, and reduced cerebral blood flow, have been implicated in cognitive decline, prompting research into interventions. Among these, Coenzyme Q10 (CoQ10), an antioxidant and metabolic stimulant, has shown promise in improving some of the underlying biological mechanisms of cognitive decline. However, not much is known about the efficacy of CoQ10 supplementation on cognition in the elderly. Therefore, the aim of this review is to explore the efficacy of CoQ10 supplementation on cognitive function. Methods: We conducted a review of animal studies and human clinical trials investigating the effect of CoQ10 supplementation on cognition in samples who were healthy or with specific diseases. Overall, twelve studies demonstrated improved cognitive function and two showed a reduction in oxidative stress in response to CoQ10 supplementation, either alone or in combination with other compounds. Out of eight human clinical trials in healthy subjects (n = 2) and disease states (n = 6), four showed evidence of a beneficial effect of CoQ10 supplementation on cognition, while two demonstrated an increase in cerebral blood flow. Disparity in the results of the clinical trials presented here is likely due to differing testing procedures, inconsistent use of cognitive assessments, and/or varying bioavailability of different preparations of CoQ10. Conclusions: There is some evidence to suggest that cognition and the biological mechanisms that regulate it are positively impacted by CoQ10 therapy. However, it is crucial to note that the literature presents mixed results, with many human clinical trials also reporting no benefit of CoQ10 supplementation on cognitive performance. To fully evaluate the benefits of CoQ10 on cognitive function in ageing and in neurodegenerative diseases, future studies are needed that target possible mechanisms and utilise a wider range of cognitive assessments. Full article

Adenosine nucleotides and polyphosphates play a significant role in biochemistry, from participating in the formation of genetic material to serving as metabolic energy currency. In this study, we examine the stability and decomposition rates of adenosine phosphates—5′-AMP, 5′-ADP, and 5′-ATP (mentioned simply as AMP, ADP and ATP hereafter)—at temperatures of 22–25 °C, 50–55 °C, 70–75 °C, and 85–90 °C, at a pH of 4, over periods of 2 and 4 days, in both saltwater and ultrapure water, under unsealed and completely dried down conditions. We found that adenosine phosphates degrade rapidly under heat and dehydration, particularly at temperatures above 25 °C. Among the three compounds, AMP is the most stable, maintaining its integrity between 22 and 55 °C, whereas ATP begins to degrade at 22–25 °C and ADP is completely decomposed at temperatures above this range. Decomposition rates were analyzed using quantitative ³¹P-NMR, based on the detection of various phosphorus-containing species. AMP primarily hydrolyzed into phosphate, pyrophosphate and even formed 2′,3′-cAMP. In contrast, the condensed adenosine phosphates (ADP and ATP) hydrolyzed to AMP, phosphate, pyrophosphate, triphosphate, 5′-AMP and, in some cases, 2′,3′-cyclic adenosine monophosphate (2′,3′-cAMP). We also investigated the formation of these compounds in the presence of N-containing additives such as thiourea, urea, imidazole, and cyanamide at temperatures between 65 and 70 °C. Among these, cyanamide and urea were particularly effective in promoting the synthesis of adenosine monophosphates (AMPs) from phosphate and adenosine. The major products observed were 2′,3′,5′-AMPs and cyclic 2′,3′-AMPs. In some experiments, adenosine diphosphate (ADP) and dimeric nucleotide species were also detected. These findings suggest that moderately heated evaporating pools could facilitate the abiotic formation of AMPs. However, such environments would likely have been unsuitable for the long-term accumulation of these compounds due to continued degradation, though there would exist some level of these nucleotides at steady state. Full article

Background/Objectives: Multiple chemical sensitivity (MCS) is a complex environmental illness characterized by intolerance to various environmental chemicals, affecting multiple organ systems. Despite its prevalence, MCS remains poorly understood, with limited recognition by the World Health Organisation amid challenges in diagnosis due to symptom heterogeneity. This study aimed to investigate the oxidative stress status in patients diagnosed with MCS compared to healthy controls, focusing on plasma and erythrocyte markers. Methods: Blood samples from 40 MCS patients and 40 controls were analyzed for lipid peroxidation (LPO), total antioxidant activity (TAA), adenosine triphosphate (ATP), and antioxidant enzyme activities, alongside glutathione cycle components. Results: Results revealed no significant differences in plasma LPO or TAA between groups, with a reduction in 61% ATP levels in MCS subjects. However, erythrocyte analysis showed reduced levels of glutathione (GSH) and total glutathione in MCS patients. Glutathione peroxidase (GPx) activity also decreased by 15% in erythrocytes of MCS patients, suggesting increased hydrogen peroxide detoxification at the expense of oxidation of GSH to glutathione disulfide (GSSG). Because glutathione reductase activity (GRd) did not change, this GSSG could not be reduced, the GSSG/GSH ratio increased by 46%, indicating heightened intracellular oxidative stress. Catalase (CAT) activity also remained unchanged (reduced by 9%, non-significant). Conclusions: These findings highlight the role of oxidative stress in MCS pathophysiology, particularly the disruption of the glutathione cycle within erythrocytes. The study underscores the need for further research into the molecular mechanisms underlying MCS to improve diagnostic criteria and therapeutic strategies. Understanding intracellular oxidative imbalances may provide insights into the systemic dysfunction observed in MCS patients. Full article

Microglia are central to neuroimmune responses and undergo dynamic structural and functional changes in models of stress and addiction, and in response to pharmacological treatments. While transcriptomic and proteomic assays provide insights into molecular profiles, morphological analysis remains a valuable proxy for assessing region-specific microglial response. However, morphological features alone often fail to capture the full complexity of microglial function, underscoring the need for standardized methods and complementary approaches. Here, we describe a standardized imaging pipeline for analyzing microglia in the nucleus accumbens core (NAcore), integrating unbiased confocal image acquisition with precise anatomical reference points. We compare two widely used image analysis platforms—IMARIS and CellSelect-3DMorph—highlighting their workflows, output metrics, and utility in quantifying microglial morphology following treatment with adenosine triphosphate (ATP). Both tools detect well described features of microglial dynamics, though they differ in automation level, analysis speed, and output types. Our findings demonstrate that both platforms provide reliable morphological data, with CellSelect-3DMorph offering a rapid, open-access alternative for high-throughput analysis. Additionally, using software-derived parameters in principal component analysis clustering has proven useful for identifying distinct subpopulations of microglia separated by their morphology. This work provides a practical framework for morphological analysis and promotes reproducibility in microglial studies under environmental and pharmacological interventions. Full article

Streptococcus mutans (S. mutans) is a cariogenic bacterium in the oral cavity that plays a significant role in plaque formation and dental caries. In previous research by our group, we showed that the endocannabinoid anandamide (AEA) has anti-bacterial and anti-biofilm activities against S. mutans. Here, we aimed to investigate its effects on S. mutans through metabolomics analyses. S. mutans was cultivated in the absence or presence of AEA at a sub-minimum inhibitory concentration (MIC), and changes in metabolites and metabolic pathways were assessed through liquid chromatography–mass spectrometry (LC-MS). Treatment of S. mutans using AEA at 10 µg/mL significantly disturbed the glycolytic flux in the bacteria, which was indicated by a reduced glucose uptake into the cell, suppression of key glycolytic intermediates, reduced acid production into the media, imbalance of NAD⁺/NADH, and decreased adenosine triphosphate (ATP) production. The disruption of carbohydrate metabolism impacts critical cellular processes, including energy production, redox balance, and biosynthetic pathways, leading to metabolic stress and impaired cellular function. These results highlight the mode of action of AEA as an antimicrobial agent. Altogether, these findings suggest that AEA has potential as a novel antimicrobial agent in the development of therapeutics against S. mutans. Full article