Induction Of Oxidative Stress Research Articles

In Vivo Evaluation of The Hepatotoxic Disorders of Tributyltin Low Doses in Male Rats

Tributyltin (TBT) has been used widely for over 60 years as a pesticidal, preservation, rodent repellent, water-repellant coating, corrosion inhibitor, flame-resistant polyester, and household product. Previous research has demonstrated the toxicity of high doses of TBT, but there is insufficient data on the effects of relevant levels of TBT on at low doses. The main goal of this thesis is to recognize the hepatotoxicity of Sublethal doses of TBT chloride (10-2000 µg/Kg body weight) in male rats for 45 days. TBT induced hepatic induction of oxidative stress as indicated by increasing levels of TBARS and 8-OHdG, induction of hepatic inflammation as indicated by the elevated NF-B levels; inhibition of hepatic expression of PGC-1α and impairment of mitochondrial biogenesis, induction of SREBP-1c and induce hepatic lipogenesis and fatty liver, and distortion of hepatic structure at histological level in a dose-dependent manner significantly at doses higher than 250 µg/kg. In conclusion: the TBT at low doses can induce hepatotoxicity at different levels.

Toxic Effect of Methyl-Thiophanate on Bombyx mori Based on Physiological and Transcriptomic Analysis

Background/Objectives: The utilization of methyl-thiophanate (MT) in vegetables and fruits is widespread due to its broad efficiency, yet its potential impact on silkworm growth remains uncertain. This study aims to examine the effects of MT on the growth of silkworms. Specifically, we assessed the weights of fifth-instar larvae that were fed mulberry leaves saturated with three concentrations (2.5, 5, and 10 mg/mL) of MT, as well as the weights of a control group. Methods: TEM was used to show the status of the silkworm midgut after MT supplementation. Oxidative stress was evaluated in the presence of MT. Furthermore, a transcriptomic sequencing experiment was conducted to investigate the mechanism through which the development of silkworms is induced by MT. Results: Our findings indicate that the supplementation of MT hindered larval growth compared to the control group, suggesting a toxic effect of MT on silkworms. The transmission electron microscopy (TEM) results show that MT supplementation induced autophagy in the silkworm midgut. MT was also found to induce oxidative stress in silkworms through the activation of reactive oxygen (ROS), superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities. Subsequent transcriptomic analysis revealed 1265 significantly differentially expressed genes (DEGs) in response to MT. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that these DEGs were associated with antioxidant defense, detoxification processes, lysosome biogenesis, and metabolic pathways. Conclusions: These findings suggest that MT toxicity in silkworm larvae is mediated through the induction of oxidative stress and alterations in metabolism. This study contributes to our understanding of the impacts of MT exposure on silkworms and provides insights into potential pesticides for use in mulberry gardens.

Selected micro- and macro-element associations with oxidative status markers in common carp (Cyprinus carpio) blood serum and ejaculate: a correlation study

ABSTRACT The aim of this study was to (1) determine complex interactions between macro- and micro-elements present in blood serum and ejaculate of common carp (Cyprinus carpio), and (2) examine the association between alterations in these macro- and micro-elements with markers of oxidative stress. Blood and ejaculate from 10 male carp were collected in the summer period on the experimental pond in Kolíňany (West Slovak Lowland). Reactive oxygen species (ROS), total antioxidant capacity (TAC), protein carbonyls (PC), and malondialdehyde (MDA) levels were measured in blood serum and ejaculate using spectrophotometric methods. The amounts of elements (Ag, Al, Ba, Co, Li, Mo, Ca, K, Na, and Mg) in all samples were quantified using inductively coupled plasma optical emission spectrophotometry. Data demonstrated significant differences in elemental concentrations between blood and ejaculate, specifically significantly higher ejaculate levels were detected for Ag, Al, Ba, Co, Li, Mo, K, and Mg. Potassium was the most abundant macro-element in the ejaculate, while sodium was the most abundant in blood serum. Among the micro-elements, Al was predominant in both types of samples. It is noteworthy that oxidative status markers including ROS, TAC, and MDA were significantly higher in ejaculate indicating the presence of oxidative stress in C. carpio reproductive tissue. The positive correlations between Mg and Ca in blood serum and ejaculate suggest these elements play a functional role in metabolic and physiological processes. In contrast, the positive correlations of Ba and Al with markers of oxidative stress indicated the association of these metals with induction of oxidative stress. Our findings provide insights into the association of metals with biomarkers of physiological function as well as adverse effects in C. carpio.

Role of Nrf2 in Epilepsy Treatment.

Oxidative stress is a consequence of the disruption of the balance between the generation of reactive nitrogen and oxygen species and the biological system's ability to neutralize those reactive products. Oxidative stress is involved in the generation of many disorders, including epilepsy, which is a prevalent chronic neurological disease that affects the lives of millions of people around the world. Epilepsy is characterized by unforeseeable and repeated seizures that can be very disturbing. Studies have reported that oxidative stress occurs before and after seizures. A transcription factor named Nuclear factor erythroid-derived 2-related factor 2 (Nrf2) controls genes related to the induction of oxidative stress and defends cells against oxidative stress. The Nrf2 protein has seven different domains, ranging from Neh1 to Neh7. Each domain is responsible for a distinctive function of this protein. Keap1 binds to Nrf2, but during oxidative stress, Nrf2 detaches from the Keap1 protein, moves to the nucleus, and binds to DNA. The result of this translocation and binding is the initiation of transcription of detoxifying genes to control the harmful effects of oxidative stress. There is some evidence of oxidative stress involvement in epilepsy. In this review, we have listed potential Nrf2-related therapeutic targets for treating and controlling epilepsy, such as Berberis alkaloids, pentoxifylline, lovastatin, progesterone, and chrysin nanoparticles. These activators were tested in animals (in vivo) and cells (in vitro), and most of these experiments showed promising results in different epilepsy models. Finally, the results have suggested that the activation of Nrf2 can be an option for controlling epilepsy.

RNAi-mediated silencing of transferrin promotes entomopathogens lethality in Odontotermes formosanus (Shiraki)

Transferrin (Tsf) is a highly conserved multifunctional protein involved in insect physiology, defense and development that has been developed as a novel RNA interference (RNAi)-based target for pest control. The function study of the Tsf gene in Odontotermes formosanu (Shiraki) was evaluated for synergistic control of this agroforestry pest with Serratia marcescens (SM1), Bacillus thuringiensis (Bt) or Beauveria bassiana (Bb). The Tsf gene of O. formosanus was identified and characterized. Real-time fluorescent quantitative PCR (qPCR) analysis demonstrated that OfTsf was most highly expressed in the male dealate of O. formosanus, and OfTsf was highly expressed in the hemolymph. OfTsf expression was considerably elevated after SM1, Bt or Bb infection. Furthermore, dsOfTsf treatment was effective in increasing the virulence of entomopathogens to O. formosanus. In addition, OfTsf expression was markedly upregulated in O. formosanus fed with oxidative stress inducers; reactive oxygen species (ROS) levels were significantly increased after dsOfTsf treatment. Therefore, OfTsf gene played an important role in defending against entomopathogen infection and antioxidant stress. Most importantly, our work suggested OfTsf as a potential RNAi target for the control of O. formosanus.

Proinflammatory Microenvironment in Adenocarcinoma Tissue of Colorectal Carcinoma.

Cancer-promoting proinflammatory microenvironment influences colorectal cancer (CRC) development. We examined the biomarkers of inflammation, intestinal differentiation, and DNA activity correlated with the clinical parameters to observe progression and prognosis in the adenocarcinoma subtype of CRC. Their immunohistology, immunoblotting, and RT-PCR analyses were performed in the adenocarcinoma and neighboring healthy tissues of 64 patients with CRC after routine colorectal surgery. Proinflammatory nuclear factor kappa B (NFκB) signaling as well as interleukin 6 (IL-6) and S100 protein levels were upregulated in adenocarcinoma compared with nearby healthy colon tissue. In contrast to nitrotyrosine expression, the oxidative stress marker 8-Hydroxy-2'-deoxyguanosine (8-OHdG) was increased in adenocarcinoma tissue. Biomarkers of intestinal differentiation β-catenin and mucin 2 (MUC2) were inversely regulated, with the former upregulated in adenocarcinoma tissue and positively correlated with tumor marker CA19-9. Downregulation of MUC2 expression correlated with the increased 2-year survival rate of patients with CRC. Proliferation-related mammalian target of rapamycin (mTOR) signaling was activated, and Ki67 frequency was three-fold augmented in positive correlation with metastasis and cancer stage, respectively. Conclusion: We demonstrated a parallel induction of oxidative stress and inflammation biomarkers in adenocarcinoma tissue that was not reflected in the neighboring healthy colon tissue of CRC. The expansiveness of colorectal adenocarcinoma was confirmed by irregular intestinal differentiation and elevated proliferation biomarkers, predominantly Ki67. The origin of the linked inflammatory factors was in adenocarcinoma tissue, with an accompanying systemic immune response.

Neuroinflammation and oxidative redox imbalance drive memory dysfunction in adolescent rats prenatally exposed to Datura Stramonium

Although there have been reports indicating that Datura Stramonium (D. stramonium) may induce anticholinergic and neuropsychiatry effects, the compound is still being used for recreational and medicinal purposes while ingestion during pregnancy has been documented. Intriguingly, minimal studies have investigated the potential neurotoxic impact of D. stramonium exposure at various stages of gestation, including its potential implication on neurophysiological well-being later in life. The present study, therefore, examined spontaneous working memory and the expression of specific neurochemicals modulating crucial neural processes in adolescent rats exposed to high and low D. stramonium doses during different stages of gestation. Pregnant rats were orally infused with 150- or 500- mg/kg/day of D. stramonium either during mid- (second week; days 8–14) or late- (third week; days 15–21) gestation, while control rats received PBS at dosing periods. Behavioral characterization of offspring between postnatal days (PD) 40 and 41 in the Y-maze revealed that D. stramonium perturbed spatial working memory in rats, although locomotor activity was generally unaltered. In addition to SOD and nitric oxide downregulation, induction of oxidative stress in the hippocampus and prefrontal cortex (PFC) of young adult rats prenatally exposed to D. stramonium was corroborated by depletion of key antioxidant regulatory elements glutathione peroxidase, glutathione reductase and catalase, which was accompanied by lipid peroxidation shown by increased MDA levels. Whereas increased expression of acetylcholinesterase and LDH was seen in adolescent rats prenatally infused D. stramonium, acetylcholine levels were downregulated in both hippocampal and PFC lysates, suggesting cholinergic and metabolic dysfunctions. Immunohistochemical labelling of GFAP and IBA-1 revealed increased expression of reactive astrocytes and microglia respectively, while the accompanying TNFα upregulation in both the hippocampus (dentate gyrus) and PFC causally linked intrauterine D. stramonium exposure with neuroinflammatory responses postnatally. Overall, our data correlated postnatal spatial working memory dysfunction evoked by D. stramonium exposure during critical stages of embryonic development to oxidative redox impairment, cholinergic disruption and neuroinflammatory perturbations in rats.

PARP-1 negatively regulates nucleolar protein pool and mitochondrial activity: a cell protective mechanism

BackgroundPoly(ADP-ribose) polymerase-1 (PARP-1) is a pan nuclear protein that utilizes NAD+ as a substrate for poly(ADP-ribosyl)ation reaction (PARylation), resulting in both auto-modification and the modification of its accepter proteins. Earlier reports suggested that several nucleolar proteins interact and colocalize with PARP-1, leading to their PARylation. However, whether PARP-1 has any role in nucleolar biogenesis and the functional relevance of such a role is still obscure.ResultsUsing PARP-1 depleted cells, we investigated the function of PARP-1 in maintaining the nucleolar morphology and protein levels under normal physiological conditions. Our results revealed that several nucleolar proteins like nucleolin, fibrillarin, and nucleophosmin get up-regulated when PARP-1 is depleted. Additionally, in line with the higher accumulation of nucleolin, stably depleted PARP-1 cells show lower activation of caspase-3, lesser annexin-V staining, and reduced accumulation of AIF in the nucleus upon induction of oxidative stress. Concurrently, PARP-1 silenced cells showed higher mitochondrial oxidative phosphorylation and more fragmented and intermediate mitochondria than the parental counterpart, suggesting higher metabolic activity for better survival.ConclusionBased on our findings, we demonstrate that PARP-1 may have a role in regulating nucleolar protein levels and mitochondrial activity, thus maintaining the homeostasis between cell protective and cell death pathways, and such cell-protective mechanism could be implicated as the priming state of a pre-cancerous condition or tumour dormancy.

N-Acetylcysteine Mitigates Oxidative Stress Induced by Transplanting Lettuce Seedlings into a DFT Hydroponic System

Oxidative stress results from an imbalance between the production and accumulation of reactive oxygen species (ROS), which can impede plant growth under various environmental stresses. While waterlogging is a well-known inducer of oxidative stress, the effects of oxidative stress on plant roots grown using the deep flow technique (DFT) hydroponic system remain poorly understood. In this study, we demonstrate that N-acetylcysteine (NAC) enhances the growth of lettuce seedlings transplanted into a DFT system. NAC application significantly improved both shoot and root growth, with the most pronounced effects observed at a concentration of 0.3 mM. Moreover, NAC mitigated the accumulation of hydrogen peroxide in roots following transplantation. It also reduced a temporary increase in lipid peroxidation and total phenolic content in both roots and shoots. These results suggest that NAC functions as an antioxidant, alleviating oxidative stress by scavenging hydrogen peroxide in the roots. Importantly, NAC’s protective effects may extend to other hydroponically grown crops, offering broader potential for reducing oxidative stress across various cultivation systems.

In silico Investigations of Al2O3 and NiO Nanoparticles Interactions with Antioxidant Enzymes of Fresh Water Fish O.mossambicus

Metal oxide nanoparticles caused hazardous to aquatic organisms, especially fishes. Nanomaterials of Aluminium and nickel oxide are applied in wide range of applications. However the toxicity of these metal oxide nanoparticles on freshwater fish is very scarce. In this study, we investigated the toxic impact of these two nanoparticles through an in silico approach. The antioxidant enzymes /proteins of SOD, CAT and GST in fish O.mossambicus were docked against Al2O3 and NiO NPs. These enzymes are responsible for oxidative metabolism of a large number of nanomaterials. Induction of oxidative stress by NPs exposure leads to inhibition of these enzymes. The targeted proteins SOD, CAT and GST were retrieved from Alpha fold database and docked with ligands. Molecular docking was conducted using CB - Dock and AutoDock Vina software tools to find the binding affinity and interaction with amino acids of each protein. The target proteins found to strongly interact with ligands through hydrogen bonding and metal coordination interactions. The lowest binding energy shows strong binding of ligands with proteins. Docking results revealed that the amino acids residues of SOD, CAT and GST enzymes are strongly bind with the Al2O3 and NiO NPs in its active site and causes inhibition of the enzymes activities. CAT exhibited lowest binding affinity of (-4.0 Kcal/mol) among all enzymes and significantly inhibited by NiO NPs, followed by GST (-3.4 Kcal/mol), likewise CAT (-1.7 Kcal/mol) and GST (-1.8 Kcal/mol) was moderately inhibited by Al2O3 NPs. SOD inhibited insignificantly by both the NPs. In conclusion, NiO NPs effectively inhibit antioxidant enzymes activity than Al2O3 NPs.

Abstract C037: Characterization of cysteine dependency implicates glutathione metabolism as a critical adaptation axis in pancreatic cancer

Abstract Pancreatic ductal adenocarcinoma (PDAC) is a deadly cancer that is highly malignant and lethal. The treatment options are limited, contributing to its high mortality rate. PDAC is known to have profound metabolic alterations that enable its growth and survival. Here, we screened for amino acid dependency in PDAC cell lines and found that the cells cluster into cysteine highly dependent, moderately dependent, and independent cells. In the highly dependent cells, cysteine deprivation reduces viability by ∼90%, which could be rescued by glutathione (GSH) supplementation. Metabolomics revealed that cysteine deprivation led to GSH depletion, suggesting an impairment in the GSH pathway machinery in these cells. Consistently, most metabolite intermediates in the GSH pathway are downregulated in the cysteine-reliant PDAC cell lines compared to cell lines that resist cysteine deprivation, and those cells showed higher sensitivity to oxidative stress induction. Bioinformatics analysis of multiple PDAC tumor gene expression datasets indicated that the GSH pathway is strongly upregulated in PDAC and correlates with proliferation, metastasis, and oncogenic KRAS upregulation. Using CRISPR/Cas9, we found that the knockout of GSH genes enhanced sensitivity to oxidative stress. Our data implicate the GSH pathway as a critical nexus to be targeted to overcome metabolic adaptation in PDAC. Citation Format: Chiamaka J Ezeh, Matthew H Ward, Chesta Jain, Verodia Charlestin, Daeho D Kim, Andrew Yang, Maya Nassif, Nneka Mbah, Peter Sajjakulnukit, Anthony Andren, Li Zhang, Marina Pasca di Magliano, Costas A Lyssiotis, Zeribe C Nwosu. Characterization of cysteine dependency implicates glutathione metabolism as a critical adaptation axis in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr C037.

Cell death and DNA damage via ROS mechanisms after applied antibiotics and antioxidants doses in prostate hyperplasia primary cell cultures.

Tumor heterogeneity results in aggressive cancer phenotypes with acquired resistance. However, combining chemical treatment with adjuvant therapies that cause cellular structure and function perturbations may diminish the ability of cancer cells to resist at chemical treatment and lead to a less aggressive cancer phenotype. Applied treatments on prostate hyperplasia primary cell cultures exerted their antitumor activities through mechanisms including cell cycle blockage, oxidative stress, and cell death induction by flow cytometry methods. A 5.37 mM Chloramphenicol dose acts on prostate hyperplasia cells by increasing the pro-oxidant status, inducing apoptosis, autophagy, and DNA damage, but without ROS changes. Adding 6.30 mM vitamin C or 622 µM vitamin E as a supplement to 859.33 µM Chloramphenicol dose in prostate hyperplasia cells determines a significant increase of ROS level for a part of cells. However, other cells remain refractory to initial ROS, with significant changes in apoptosis, autophagy, and cell cycle arrest in G0/G1 or G2/M. When the dose of Chloramphenicol was increased to 5.37 mM for 6.30 mM of vitamin C, prostate hyperplasia cells reacted by ROS level drastically decreased, cell cycle arrest in G2/M, active apoptosis, and autophagy. The pro-oxidant action of 1.51 mM Erythromycin dose in prostate hyperplasia cell cultures induces changes in the apoptosis mechanisms and cell cycle arrest in G0/G1. Addition of 6.30 mM vitamin C to 1.51 mM Erythromycin dose in hyperplasia cell cultures, the pro-oxidant status determines diminished caspase 3/7 mechanism activation, but ROS level presents similar changes as Chloramphenicol dose and cell cycle arrest in G2/M. Flow cytometric analysis of cell death, oxidative stress, and cell cycle are recommended as laboratory techniques in therapeutic and diagnostic fields.

Molecular Events in Response to Triclosan-Induced Oxidative Stress in CRISPR/Cas9-Mediated p53-Targeted Mutants in Daphnia magna.

Triclosan (TCS), a widely used antimicrobial agent, has been implicated in the oxidative stress induction and disruption of cellular processes in aquatic organisms. As TCS is ubiquitous in the aquatic environment, many previous studies have documented the effects of exposure to TCS on aquatic organisms. Nevertheless, most of the research has concentrated on the molecular and physiological responses of TCS, but there are still limited studies on the function of specific genes and the consequences of their absence. In this study, we focused on p53, a gene that is crucial for molecular responses such as autophagy and apoptosis as a result of TCS exposure. In order to ascertain the role and impact of the p53 gene in TCS-induced molecular responses, we examined the molecular responses to TCS-induced oxidative stress in wild-type (WT) and CRISPR/Cas9-mediated p53 mutant (MT) water fleas. The result has been accomplished by examining changes in molecular mechanisms, including in vivo end points, enzyme activities, adenosine triphosphate release rate, and apoptosis, to determine the role and impact of the p53 gene on TCS-induced molecular responses. The results indicated that the sensitivity of MT water fleas to TCS was greater than that of WT water fleas; however, the difference in sensitivity was significant at short exposures within 48 h and decreased toward 48 h. Accordingly, when we confirmed the oxidative stress after 24 h of exposure, the oxidative stress to TCS exposure was stronger in the MT group, with an imbalance of redox. To identify the mechanisms of tolerance to TCS in WT and MT Daphnia magna, we checked mitochondrial and ER-stress-related biomarkers and found an increase in apoptosis and greater sensitivity to TCS exposure in the MT group than in the WT. Our results suggest that the absence of p53 caused alterations in molecular processes in response to TCS exposure, resulting in increased sensitivity to TCS, and that p53 plays a critical role in response to TCS exposure.

Triggering multiple modalities of cell death via dual-responsive nanomedicines to address the narrow therapeutic window of β-lapachone

The clinical translation of the anticancer drug β-lapachone (LAP) has been limited by the narrow therapeutic window. Stimuli-responsive anticancer drug delivery systems have gained tremendous attention in recent years to alleviate adverse effects and enhance therapeutic efficacy. Here, we report a dual pH- and enzyme-responsive nanocarrier to address the above issue of LAP. In detail, the epigallocatechin gallate (EGCG) and ferric ions were employed to engineer nanoscale ferric ion-polyphenol complexes where LAP was physically encapsulated therein. The coordination bond between Fe3+ and the catechol moiety of EGCG was sensitive to the low pH, enabling the triggered cargo release in the acidic endosomes/lysosomes. Afterward, the released LAP was activated by the overexpressed NAD(P)H: quinone oxidoreductase 1 (NQO1) and ferroptosis suppressor protein 1 (FSP1) in the tumor cells, followed by the generation of reactive oxygen species (ROS), and the induction of oxidative stress and apoptotic cell death. Meanwhile, EGCG could upregulate gasdermin E (GSDME), and ferric ions were involved in the Fenton reaction. Hence, EGCG and ferric ions could sensitize the toxicity of LAP through the induction of multiple cell death pathways (e.g., pyroptosis, ferroptosis, apoptosis, and necroptosis). The current work enlarged the LAP’s therapeutic window via controlled cargo release and vehicle sensitization.

Cellular toxicity of calcium propionate in human lymphocyte.

Calcium propionate is the chemical substance added to food in order to prolong the shelf-life of factory made foods by inhibiting the development of bacteria, fungi and other microorganisms. The objective of this study was to investigate the ability of calcium propionate to induce cytotoxic and genotoxic effects in lymphocytes. Oxidative stress induction by calcium propionate was also studied. Four concentrations of calcium propionate (0.5, 1.0, 1.5 and 2.0 mg/ml) were applied in lymphocytes for 24 and 48 h treatment. It studied cytotoxic and genotoxic effects by MTT assay, chromosome culture technique, and micronucleus assay. Oxidative stress induction was studied by superoxide dismutase (SOD) activity assay. The results showed that lymphocyte viability was decreased significantly by calcium propionate at 1.5 and 2.0 mg/ml (p < 0.05). Calcium propionate induced chromosome aberration at 1.0, 1.5 and 2.0 mg/ml and sister chromatid exchange at 1.5 and 2.0 mg/ml (p < 0.05). It induced micronucleus formation at 0.5, 1.0, 1.5 and 2.0 mg/ml (p < 0.05). The calcium propionate concentrations of 0.5 - 1.0 mg/ml and 1.5 - 2.0 mg/ml could reduce SOD activity inhibition (p < 0.05). Calcium propionate induced oxidative stress in lymphocytes. It can be concluded that calcium propionate induces genotoxic risk and oxidative stress in lymphocytes. Based on this study and the positive results, consumers should be made aware that calcium propionate should be considered a genotoxic compound. The awareness of food preservative usage and the educational program must take place frequently for good human health in the community.

Super-Resolution Microscopic Imaging of Lipid Droplets in Living Cells via Carbonized Polymer Dot-Based Polarity-Responsive Nanoprobe.

Lipid droplets (LDs) are dynamic subcellular organelles that participate in various physiological processes, and their abnormality can also lead to various diseases. Tracing the dynamics of LDs in living cells will be valuable for understanding cell physiological states. Here, we employed a structured light illumination super-resolution imaging assisted with a carbonized polymer dot (CPD)-based fluorescence nanoprobe to track the travel paths of LDs and other organelles. The CPDs we developed are highly biocompatible with living cells and exhibit a highly sensitive response to solvent polarity, allowing for high specificity in staining LDs in living cells. Aided by these nanoprobes, we successfully observed many real-time LD-involved dynamics in living cells, such as intracellular LD interactions, communications with other organelles, and dynamic behaviors under external stimuli (oxidative stress inducer). These studies deepen our comprehension of the physiological role of LDs and drive the advancement of super-resolution fluorescent probes.

Impeding microbial biofilm formation and Pseudomonas aeruginosa virulence genes using biologically synthesized silver Carthamus nanoparticles

Long-term antibiotic treatment results in the increasing resistance of bacteria to antimicrobials drugs, so it is necessary to search for effective alternatives to prevent and treat pathogens that cause diseases. This study is aimed for biological synthesis of silver Carthamus nanoparticles (Ag-Carth-NPs) to combat microbial biofilm formation and Pseudomonas aeruginosa virulence genes. Ag-Carth-NPs are synthesized using Carthamus tenuis aqueous extract as environmentally friendly method has no harmful effect on environment. General factorial design is used to optimize Ag-Carth-NPs synthesis using three variables in three levels are Carthamus extract concentration, silver nitrate concentration and gamma radiation doses. Analysis of response data indicates gamma radiation has a significant effect on Ag-Carth-NPs production. Ag-Carth-NPs have sharp peak at λ max 425 nm, small and spherical particles with size 20.0 ± 1.22 nm, high stability up to 240 day with zeta potential around − 43 ± 0.12 mV, face centered cubic crystalline structure and FT-IR spectroscopy shows peak around 620 cm−1 that corresponding to AgNPs that stabilized by C. tenuis extract functional moiety. The antibacterial activity of Ag-Carth-NPs against pathogenic bacteria and fungi was determined using well diffusion method. The MIC values of Ag-Carth-NPs were (6.25, 6.25, 3.126, 25, 12.5, 12.5, 25 and 12.5 µg/ml), MBC values were (12.5, 12.5, 6.25, 50, 25, 25, 50 and 25 µg/ml) and biofilm inhibition% were (62.12, 68.25, 90.12, 69.51, 70.61, 71.12, 75.51 and 77.71%) against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Candida tropicalis and Candida albicans respectively. Ag-Carth-NPs has bactericidal efficacy and significantly reduced the swarming, swimming motility, pyocyanin and protease production of P. aeruginosa. Furthermore, P. aeruginosa ToxA gene expression was significantly down regulated by 81.5%, while exoU reduced by 78.1%, where lasR gene expression reduction was 68%, while the reduction in exoU was 66% and 60.1% decrease in lasB gene expression after treatment with Ag-Carth-NPs. This activity is attributed to effect of Ag-Carth-NPs on cell membrane integrity, down regulation of virulence gene expression, and induction of general and oxidative stress in P. aeruginosa. Ag-Carth-NPs have no significant cytotoxic effects on normal human cell (Hfb4) but have IC50 at 5.6µg/mL against of HepG-2 cells. Limitations of the study include studies with low risks of silver nanoparticles for in vitro antimicrobial effects and its toxicity.

Oxidative lipid damage by naringenin selectively sensitizes chronic myeloid leukemia cell lines and patient samples to Bcr-Abl tyrosine kinase inhibitors

Chronic myeloid leukemia (CML) treatment with Bcr-Abl tyrosine kinase inhibitors (TKIs) has significantly improved patient outcomes, yet challenges such as drug resistance and persistence of leukemic stem cells persist. This study explores the potential of naringenin, a natural flavonoid, to enhance the efficacy of Bcr-Abl TKIs in CML therapy. We showed that naringenin reduces viability of a panel of CML cell lines regardless of varying cellular origin and genetic mutations, and acts synergistically with dasatinib and ponatinib. Importantly, naringenin is effective in targeting blast crisis CML CD34+ cells by decreasing their colony formation, self-renewal and viability. Compared to CML, naringenin is significantly less effective against normal bone marrow (NBM) counterparts. In addition, naringenin significantly enhances the inhibitory effects of dasatinib in CML but not NBM CD34+ cells. Mechanism studies showed that naringenin's inhibitory effects were associated with the induction of oxidative stress and lipid damage, as evidenced by increased reactive oxygen species (ROS) and malondialdehyde (MDA) levels. Notably, naringenin upregulated genes related to mitochondrial biogenesis while downregulating antioxidant defense genes. Pretreatment with α-tocopherol, which inhibits lipid-mediated ROS production, completely abolished the ROS increase and restored cell viability, indicating that lysosomal lipid peroxidation plays a crucial role in naringenin's mechanism of action. In a CML xenograft mouse model, the combination of naringenin and dasatinib resulted in remarkably more tumor growth suppression compared to single drug alone. Importantly, this combination was well-tolerated, with no adverse effects on body weight observed. These findings suggest that naringenin, by inducing oxidative lipid damage, enhances the anti-leukemic effects of Bcr-Abl TKIs, offering a promising therapeutic strategy for CML.

A Study to Evaluate the Effect of Alpha-Lipoic Acid on Neuropathic Symptoms in Diabetic Neuropathy Patients on Gabapentin or Pregabalin.

Introduction Chronic hyperglycemia is a key factor in the development of diabetic peripheral neuropathy (DPN), contributing significantly to the progression of this condition through the induction of oxidative stress. Elevated blood glucose levels lead to increased production of reactive oxygen species (ROS), which cause damage to neuronal cells and exacerbate neuropathic symptoms. Alpha-lipoic acid (ALA) is a potent antioxidant that neutralizes ROS and reduces oxidative damage. By addressing the mechanisms of neuropathy, ALA mitigates the effects of chronic hyperglycemia through antioxidant regeneration, inflammation reduction, and endothelial function improvement. As a result, ALA is emerging as a promising intervention for managing oxidative stress and inflammation in DPN. Methods Prospective, double-blind, placebo-controlled study, a total of 52 DPN subjects on gabapentin or pregabalin were randomly assigned to either ALA 600 mg oral once daily or placebo oral once daily for 12 weeks. Treatment outcome was assessed using vibration perception threshold (VPT), Neuropathy Total Symptom Score 6 (NTSS-6), quality of life (QOL) assessed by 12-Item Short Form Health Survey (SF-12) Questionnaire, oxidative stress biomarkers (malondialdehyde (MDA), nitric oxide (NO), and glutathione (GSH)) and inflammatory biomarker (high-sensitivity C-reactive protein (hs-CRP)) at baseline and every visit (four, eight, and 12 weeks). Neuron-specific enolase (NSE) levels were done at baseline and 12 weeks. Glycated hemoglobin (HbA1C) and safety parameters were done at screening and 12 weeks. Compliance with the study medication was assessed by pill count. Reported adverse drug reactions were recorded. Results A total of 52 subjects (males = 22; females = 30) with a mean age of 55.63 ± 7.5 years were randomized to receive either ALA or placebo. A significant reduction in VPT was observed with ALA at 12 weeks compared to baseline (from 26.92 ± 3.58 to 22.14 ± 1.94; p < 0.0001). Secondary parameters like NTSS-6, QOL by SF-12 questionnaire, NSE levels, oxidative biomarkers levels, and inflammatory biomarkers showed significant improvement with ALA compared to placebo. No serious adverse events were reported. Conclusion ALA demonstrated a protective effect against DPN by its antioxidant and anti-inflammatory effects and was found to have good safety.

Hyperhomocysteinemia-Induced Alterations in Protein Expression and Oxidative Stress Parameters in Rat Heart

Hyperhomocysteinemia (HHcy) is considered an independent risk factor of cardiovascular diseases. Among the proposed mechanisms underlying homocysteine toxicity are altered protein expression and induction of oxidative stress. In the present study, we explored protein abundance and parameters related to oxidative stress in heart homogenates of rats exposed to chronic mild HHcy. Using two-dimensional gel electrophoresis followed by MALDI-TOF/TOF mass spectrometry 22 altered proteins (6 upregulated and 14 downregulated) were identified. For eight proteins the altered abundances were validated by Western blot analysis. Identified proteins are primarily involved in energy metabolism (mainly enzymes of glycolysis, pyruvate dehydrogenase complex, citric acid cycle, and ATP synthase), cardiac muscle contraction (α-actin and myosin light chains), stress response (heat-shock protein β1 and αB-crystallin) and antioxidant defense (glutathione peroxidase 1). Diminished antioxidant defense was confirmed by decreases in total antioxidant capacity and GSH/GSSG ratio. Consistent with the decline in enzymatic and non-enzymatic antioxidant defense the protein oxidative modification, as determined by tyrosine nitration, was significantly increased. These findings suggest that both, altered protein expression and elevated oxidative stress contribute to cardiovascular injury caused by HHcy.