Oxidative Stress Conditions Research Articles

Musa basjoo Sieb polysaccharide improves inflammation in RAW264.7 cells and zebrafish colitis

Musa basjoo Sieb polysaccharide (MBSP) is the active ingredient of Musa basjoo Sieb. The purpose of this study was to preliminarily characterize the structural properties of MBSP, and to explore its ameliorating effect LPS-induced macrophage RAW264.7 inflammation and its therapeutic effect on DSS-induced zebrafish colitis. The data indicated that the average molecular weight of MBSP was 393.4 kDa, which was an acidic heteropolysaccharide composed of seven monosaccharides. In the LPS-induced macrophage RAW264.7 inflammation model, we found that MBSP significantly reduced the level of inflammatory mediators (p < 0.001). The release of NO, IL-6, and TNF-α was reduced by 48.21%, 65.45%, and 86.49%, respectively. And MBSP modulated intracellular ROS levels. This might be related to the activation of NF-κB/MAPK signaling pathway by MBSP. Additionally, MBSP significantly improved the oxidative stress status of UC zebrafish. SOD, CAT, GSH-Px were increased by 50.58%, 39.07% and 65.86%, and MDA, MPO were decreased by 26.00% and 50.98%, respectively (p < 0.001). Meanwhile, MBSP was also found to be effective in improving UC by activating the TLR4/NF-κB/MAPK signaling pathway in vivo. In conclusion, the present study characterized the structural properties of MBSP from various aspects, confirmed the anti-inflammatory effect of MBSP in vitro, and further verified the in vivo anti-inflammatory activity of MBSP using UC zebrafish as an animal model. The results of this study provide a reference for the development and application of MBSP in the field of treating inflammatory diseases.

Nutraceutical Features of the Phycobiliprotein C-Phycocyanin: Evidence from Arthrospira platensis (Spirulina).

Arthrospira platensis, commonly known as Spirulina, is a photosynthetic filamentous cyanobacterium (blue-green microalga) that has been utilized as a food source since ancient times. More recently, it has gained significant popularity as a dietary supplement due to its rich content of micro- and macro-nutrients. Of particular interest is a water soluble phycobiliprotein derived from Spirulina known as phycocyanin C (C-PC), which stands out as the most abundant protein in this cyanobacterium. C-PC is a fluorescent protein, with its chromophore represented by the tetrapyrrole molecule phycocyanobilin B (PCB-B). While C-PC is commonly employed in food for its coloring properties, it also serves as the molecular basis for numerous nutraceutical features associated with Spirulina. Indeed, the comprehensive C-PC, and to some extent, the isolated PCB-B, has been linked to various health-promoting effects. These benefits encompass conditions triggered by oxidative stress, inflammation, and other pathological conditions. The present review focuses on the bio-pharmacological properties of these molecules, positioning them as promising agents for potential new applications in the expanding nutraceutical market.

Innovative Molecularly Imprinted Electrochemical Sensor for Selective Nanomolar Detection of the Anti-COVID-19 Medication Molnupiravir

For the first time, a green ecofriendly approach is applied, to develop a molecularly imprinted polymer (MIP) electrochemical for the assay of the oral anti-viral molnupiravir (MLN) in various matrices as confirmed by referring to analytical eco-scale, green analytical procedure index (GAPI), Raynie and Driver, analytical greenness metric (AGREE), and national environmental index (NEMI). AuNPs were chronoamperometry deposited on the glassy carbon electrode surface (GCE) using 0.01 gm ml−1 gold solution (HAuCl4.3H2O) in 0.5 M H2SO4 at a constant potential of −0.4 V for 60 s. Then, the MIP was created by electropolymerizing OPDA on the surface of AuNPs GCE using cyclic voltammetry in the potential range between −0.5 and +1 V for 15 cycles (scan rate: 50 mV.s−1). To optimize the sensor’s performance, factors such as number of electropolymerization cycles, template: monomer ratio, binding and rebinding time, pH of rebinding buffer, extraction solvent, deposition time of Au nanoparticles, order of deposition on the electrode; surface, as well as differential pulse voltammetry parameters including accumulation potential and time, and potential step, were all investigated. The detection limit was set at 0.00098 ng ml−1 (3 × 10−12M) and the linearity range of MLN was between 0.033 ng ml−1 and 164 ng ml−1 (1 × 10−10–5 × 10−7 M). The MIP sensor was employed for MLN determination in its pharmaceutical product (capsules), spiked human plasma, and human urine samples with mean recovery % ± mean relative standard deviation (RSD) (99.82% ± 0.53), (99.88% ± 0.62), and (97.90% ± 0.70), respectively. The investigated sensor provided good reproducibility, repeatability, and durability. Acceptable selectivity regarding MLN when mixed with structurally comparable compounds was proved with mean recovery % ± mean RSD (97.10% ± 0.03). Additionally, when molnupiravir was exposed to oxidative, hydrolytic, and thermal stress conditions, good results in stability-indicating studies served as an indicator of sensor selectivity. The developed electrode is coupled with a portable potentiostat, making it a promising point-of-care diagnostic platform for on-site measurements.

Interplay between mistranslation and oxidative stress in Escherichia coli.

Mistakes in translation are mostly associated with toxic effects in the cell due to the production of functionally aberrant and misfolded proteins. However, under certain circumstances mistranslation can have beneficial effects and enable cells to preadapt to other stress conditions. Mistranslation may be caused by mistakes made by aminoacyl-tRNA synthetases, essential enzymes that link amino acids to cognate tRNAs. There is an Escherichia coli strain expressing isoleucyl-tRNA synthetase mutant variant with inactivated editing domain which produces mistranslated proteomes where valine (Val) and norvaline (Nva) are misincorporated into proteins instead of isoleucine. We compared this strain with the wild-type to determine the effects of such mistranslation on bacterial growth in oxidative stress conditions. When the cells were pre-incubated with 0.75 mmol/L Nva or 1.5 mmol/L Val or Nva and exposed to hydrogen peroxide, no beneficial effect of mistranslation was observed. However, when the editing-deficient strain was cultivated in medium supplemented with 0.75 mmol/L Val up to the early or mid-exponential phase of growth and then exposed to oxidative stress, it slightly outgrew the wild-type grown in the same conditions. Our results therefore show a modest adaptive effect of isoleucine mistranslation on bacterial growth in oxidative stress, but only in specific conditions. This points to a delicate balance between deleterious and beneficial effects of mistranslation.

Combined analysis of the effects of hypoxia and oxidative stress on DNA methylation and the transcriptome in HTR-8/SVneo trophoblast cells.

The alterations in DNA methylation and transcriptome in trophoblast cells under conditions of low oxygen and oxidative stress have major implications for pregnancy-related disorders. However, the exact mechanism is still not fully understood. In this study, we established models of hypoxia (H group) and oxidative stress (HR group) using HTR-8/SVneo trophoblast cells and performed combined analysis of genome-wide DNA methylation changes using reduced representation bisulphite sequencing and transcriptome expression changes using RNA sequencing. Our findings revealed that the H group exhibited a higher number of differentially methylated genes and differentially expressed genes than the HR group. In the H group, only 0.90% of all differentially expressed genes displayed simultaneous changes in DNA methylation and transcriptome expression. After the threshold was expanded, this number increased to 6.29% in the HR group. Notably, both the H group and HR group exhibited concurrent alterations in DNA methylation and transcriptome expression within Axon guidance and MAPK signalling pathway. Among the top 25 differentially methylated KEGG pathways in the promoter region, 11 pathways were commonly enriched in H group and HR group, accounting for 44.00%. Among the top 25 KEGG pathways in transcriptome with significant differences between the H group and HR group, 10 pathways were consistent, accounting for 40.00%. By integrating our previous data on DNA methylation from preeclamptic placental tissues, we identified that the ANKRD37 and PFKFB3 genes may contribute to the pathogenesis of preeclampsia through DNA methylation-mediated transcriptome expression under hypoxic conditions.

Liver epigenomic signature associated with chronic oxidative stress in a mouse model of glutathione deficiency

Oxidative stress is intimately involved in the pathogenesis of fatty liver disease (FLD). A major factor contributing to oxidative stress is the depletion of the ubiquitous antioxidant glutathione (GSH). Unexpectedly, chronic GSH deficiency renders glutamate-cysteine ligase modifier subunit (Gclm)-null mice protected from fatty liver injuries. Epigenetic regulation serves as an important cellular mechanism in modulating gene expression and disease outcome in FLD, although it is not well understood how systemic redox imbalance modifies the liver epigenome. In the current study, utilizing the Gclm-null mouse model, we aimed to elucidate redox-associated epigenomic changes and their implications in liver stress response. We performed high-throughput array-based DNA methylation profiling (MeDIP array) in 22,327 gene promoter regions (from −1300 bp to +500 bp of the Transcription Start Sites) in the liver and peripheral blood cells. Results from the MeDIP array demonstrate that, although global methylation enrichment in gene promoters did not change, low GSH resulted in prevalent demethylation at the individual promoter level. Such an effect likely attributed to a declined availability of the methyl donor S-adenosyl methionine (SAM) in Gclm-null liver. Functional enrichment analysis of liver target genes is suggestive of a potential role of epigenetic mechanisms in promoting cellular survival and lipid homeostasis in Gclm-null liver. In comparison with the liver tissue, MeDIP array in peripheral blood cells revealed a panel of 19 gene promoters that are candidate circulating biomarkers for hepatic epigenomic changes associated with chronic GSH deficiency. Collectively, our results provided new insights into the in vivo interplay between liver redox state and DNA methylation status. The current study laid the groundwork for future epigenetic/epigenomic investigations in experimental settings or human populations under conditions of liver oxidative stress induced by environmental or dietary challenges.

Antioxidative and Anti-inflammatory Effects of Plant-derived Hypoglycemic Medicines: An In vivo/In vitro Systematic Review.

The activation of oxidative stress and inflammatory conditions has been associated with acceleration in diabetes (DM) onset and complications. Despite various anti-DM medications, there is a growing trend to discover inexpensive and effective treatments with low adverse effects from plants as one of the promising sources for drug development. This study aimed to systematically investigate the simultaneous anti-inflammatory and antioxidant effects of plant-derived hypoglycemic medicines in diabetic experimental models. The search terms consisted of "diabetes", "herbal medicine", "antioxidant", "Inflammatory biomarker", and their equivalents among PubMed, Scopus, Web of Science, and Cochrane Library databases up to 17 August 2021. Throughout the search of databases, 201 eligible experimental studies were recorded. The results showed that the most commonly assessed inflammatory and oxidative stress biomarkers were tumor necrosis factor (TNF)-α, interleukin (IL) 6, IL-1β, IL-10, malondialdehyde (MDA), and nitric oxide (NO). The activity of antioxidant enzymes, including superoxide dismutase (SOD), glutathione (GSH), and catalase (CAT) were assessed in the present review. Among herbal treatments, Trigonella foenum-graecum L., Centella asiatica (L.) Urb., Vitis vinifera L., and Moringa oleifera Lam. were most commonly used for diabetic complications. Due to the dispersion of the treatments, meta-analysis was not applicable. Our findings showed that the application of different plant-derived hypoglycemic treatments in animal models improved diabetes and its complications, as well as modulated concomitant inflammatory and oxidative stress biomarkers. These findings suggest that plant-based antidiabetic medicines and food supplements have the potential to manage diabetes and its complications.

Development and validation of stability-indicating assay method and identification of force degradation products of glucagon-like peptide-1 synthetic analog Exenatide using liquid chromatography coupled with Orbitrap mass spectrometer.

Exenatide is a synthetic glucagon-like peptide 1 analog, widely used in the management of type 2 diabetes mellitus. The stability of pharmaceutical products is significantly impacted by various environmental stress conditions. The present study reports the development of a validated reverse-phase high-performance liquid chromatography (RP-HPLC) stability-indicating method for the identification of force degradation products (DPs) of synthetic glucagon-like peptide-1 analog Exenatide using UHPLC-Orbitrap fusionTM mass spectrometer. Force degradation studies were performed by subjecting Exenatide to various stress conditions, such as hydrolytic, oxidative, photolytic and thermal to investigate the stability indicating ability of the method. Significant degradation was observed during acidic, oxidative, photolytic and thermal stress conditions. Exenatide and its major DPs identification and characterization were demonstrated by employing LC-HRMS and MS/MS method. In total, five major stress DPs were characterized, and their fragmentation pathway was proposed using MS/MS studies. Finally, the proposed RP-HPLC method was validated as per ICH guidance.

A novel polysaccharide of Undaria pinnatifida: Structural characterization, carboxymethylation and hypoglycemic activity in vivo

A new polysaccharide (UPP) was isolated and purified from Undaria pinnatifida with a low molecular weight of 8.34 kDa. The UPP was modified to obtain carboxymethylated polysaccharide (CM-UPP). UPP and CM-UPP were characterized by FT-IR, Ion chromatography, Congo red, DSC, SEM and NMR, and their hypoglycemic activity in vivo was investigated using the STZ-induced diabetes mice model. UPP mainly contained Ara, Gal, Glc and Man at a molar ratio of 0.18: 0.13: 2.06: 1, and CM-UPP added two alduronic acids including GalA and GlcA. CM-UPP had been prepared successfully and the carboxymethylation reaction might occur partially at the C-2 and C-4 position. There were no triple helix structures in both UPP and CM-UPP. In vivo hypoglycemic activity studies showed that both UPP and CM-UPP could alleviate symptoms of high blood glucose, improve blood lipid level, oxidative stress state and liver glycolytic enzyme activity in diabetic mice. Under the same dose, CM-UPP showed a more significant effect than UPP. These results indicated that carboxymethylation was an effective way to enhance the hypoglycemic activitiy of polysaccharide. UPP and CM-UPP could be potential therapeutic agents of T2DM.

Ameliorative Effect of Pigeon Pea Leaves and Ginger Extract on Oxidative Stress Condition in Kidney and Liver of Experimental Diabetic Rats

Ameliorative Effect of Pigeon Pea Leaves and Ginger Extract on Oxidative Stress Condition in Kidney and Liver of Experimental Diabetic Rats

Characterization of Therapeutic Antibody Charge Heterogeneity Under Stress Conditions by Microfluidic Capillary Electrophoresis Coupled with Mass Spectrometry

Therapeutic antibodies are a major class of biopharmaceutics that are applied in disease treatment because of their many advantages, including high specificity and high affinity to molecular targets. Between their production and administration, therapeutic antibodies are exposed to multiple stress conditions. Forced degradation and stress stability studies are conducted to simulate the risk of degradation and the effects of these stresses, thereby enhancing understanding of the drug product to support strategies to mitigate the impact from stressed conditions. These types of studies are also routinely conducted to evaluate product comparability when major process changes are implemented during the production. Charge variant analysis helps understand the changes in the electrostatic environment of biotherapeutics and can uncover underlying molecular level alterations associated with charge variants. Herein, we used ZipChip native capillary electrophoresis-mass spectrometry (nCE-MS) to elucidate the changes in charge variant profiles at the molecular level. In two case studies under thermal stress conditions, we observed that charge variants arose from both post-translational modifications (including deamidation, oxidation, and pyroglutamate formation) and sequence truncations at the hinge regions. Under oxidative stress conditions, oxidation was found to be the major contributor to the changes in the charge variant profiles. Under pH stress conditions, the changes in the charge variant profile were due to increased levels of deamidation, oxidation, and pyroglutamate formation. ZipChip nCE-MS analysis enables identification of charge variant species under various stress conditions, thus supporting process and formulation development of biotherapeutics.

#2432 DNA damage analysis with oxidative stress index in patients developing contrast nephropathy after coronary angiography

Abstract Background and Aims Contrast Nephropathy (KN) is acute kidney injury that occurs after exposure to contrast material. Advanced age, chronic kidney disease, heart failure, anemia, hypertension, diabetes mellitus, and hyperuricemia have been reported in the literature to increase the risk of KN development. In addition, oxygen radicals have an important place in the pathogenesis of contrast nephropathy development. Specific biomarkers are used in clinics to determine oxidative stress and antioxidant status. In particular, measurement of total antioxidant status (TAS) and total oxidant status (TOS) can provide useful information about an individual's overall serum antioxidative status. The ratio of TOS to TAS level gives the oxidative stress index (OSI). One of the most investigated metabolites for DNA damage is 8-hydroxy-2-deoxyguanosine (8-OhdG), which is accepted as a biomarker of oxidative damage to DNA. In our study, we planned to investigate DNA damage and important predisposing factors for contrast nephropathy using oxidative stress markers. Method Between February 2021 and April 2022, blood was taken from 191 patients who underwent coronary angiography (CAG) by the Cardiology Department at ESOGU before CAG and at 72 hours after CAG. Patients who met the criteria for contrast nephropathy development (those with a serum creatinine increase of 25% or more compared to baseline or an increase of 0.5 mg/dl or more) were included as the case group in our study. Our study is a prospective study consisting of 85 cases and 106 controls, totaling 191 participants. TAS, TOS, OSI, and 8-OhDG levels were compared between both groups and before-after procedure. This study was supported by ESOGU BAP project number 1621. Results The mean age of the developing group was calculated as 68.13 ± 10.37, while it was calculated as 61.03 ± 9.97 in the control group (p = 0.00). In the case group, there were 30 (35.3%) without heart failure, while there were 75 (70.8%) without heart failure in the control group (p = 0.00). In univariate analyzes, we found that advanced age and heart failure were the most important indicators for KMN. When we compared those with and without KMN, we saw that TOS values decreased less in the group with post-angiography nephropathy than in those without it. We found a positive correlation between uric acid and TAS. In the KMN group, it was found that there was a statistically significant increase in TAS value after CAG, while there was a decrease in OSI value, and no statistically significant difference was found for TOS and 8-OhDG. In patients who underwent percutaneous intervention, it was found that there was a statistically significant increase in TAS value after CAG, while TOS and OSI decreased, and no significant difference was found for 8-OhDG. Conclusion Although previous studies have shown that oxidative stress plays an important role in contrast nephropathy pathogenesis, a strong relationship between KMN development and oxidative stress could not be found in our study. Our study is the first study to examine DNA damage with 8-OhDG in KMN, and it has been shown that there is no increase in DNA damage with KMN development. It is thought that one of the reasons leading to this result may be that contrast agents used today are selected as less toxic substances used in less volumes during processing, which may prevent DNA damage from occurring. Another result reached in our study is that TAS level increased in patients undergoing percutaneous intervention, while TOS and OSI decreased; that is, revascularization can reduce TOS and ultimately oxidative status.”

#1591 Analysis of microvesicles and their relationship with oxidative stress as biomarkers of cardiovascular damage in chronic kidney disease

Abstract Background and Aims Chronic kidney disease (CKD) is associated with a higher incidence of cardiovascular diseases. These diseases are preceded by endothelial dysfunction and alterations in coagulation systems. One of the challenges of research in this field is the search for diagnostic and predictive biomarkers of cardiovascular pathology. Our hypothesis is that endothelial dysfunction and cardiovascular risk could be evidenced by the increased release of endothelial microvesicles (EMVs) and platelet microvesicles (PMVs), as well as a greater expression of tissue factor (TF). We also propose a relationship between the release of microvesicles and the oxidative stress observed in these patients. The aim of the study was to analyze the levels of MVEs and MVPs, the expression of FT and its relationship with pro-oxidant and antioxidant parameters as possible early biomarkers of cardiovascular and thrombotic risk in different stages and treatments of CKD. Method It is a cross-sectional study consisted of 116 patients (40 with advanced chronic disease (CKD), 40 undergoing hemodialysis (HD) and 36 undergoing peritoneal dialysis (PD)), as well as 17 healthy subjects (CT). Microvesicle phenotyping was performed by direct immunofluorescence and flow cytometry in plasma isolated by centrifugation from peripheral blood. The analysis of oxidative stress parameters was carried out by spectrophotometry, after isolation of plasma and mononuclear and polymorphonuclear leukocytes by means of a ficoll gradient. Results Patients on PD have higher plasma PMVs levels (p = 0.0005 vs ACKD and p = 0,0002 vs HD), while those on HD and PD have higher levels of EMVs compared to ACKD (p = 0,0001 and p = 0,0125, respectively). FT expression in PMVs is lower in HD (p &amp;lt; 0.0001 vs ACKD) and FT expression in EMVs is lower in HD and PD compared to ACKD (p = 0,0004 and p= 0.0033, respectively) (Fig. 1). Patients with CKD have a higher activity of pro-oxidant enzymes (XO, GPx), as well as high levels of lipid peroxidation by-products (TBARS-MDA) and lower activity of antioxidant enzymes (GSH, CAT, SOD) at the plasma and cellular level. A positive relationship has been observed between pro-oxidant parameters and the release of MVP and MVE, as well as the expression of FT (Table 1). Patients with cardiovascular events during the years of follow-up have higher levels of FT expressed in microvesicles, and long-term survival decreases in patients with elevated FT levels. Conclusion Patients with CKD have higher plasma levels of PMVs and EMVs and higher FT expression, related to a state of oxidative stress. These parameters could be considered as biomarkers for predicting, diagnosing, and prognostic cardiovascular events in CKD.

Dietary supplementation with quercetin: an ideal approach for improving meat quality and oxidative stability of broiler chickens

This study aimed to improve the eating quality of yellow-feathered broiler chicks by feeding them corn-soybean meal diets supplemented with 250, 500, and 1,000 mg/kg quercetin. we examined the impact of varying doses of dietary quercetin on the sensory quality of chicken breast meat as well as on the antioxidant enzymes, antioxidant-related signaling molecules, structure and thermal stability of myofibrillar protein (MPs), and microstructure of myogenic fibers in the meat during 24 h of postslaughter aging. Additionally, we investigated the potential correlations among antioxidant capacity, MP structure, and meat quality parameters. The results indicated that dietary supplementations with 500 and 1,000 mg/kg quercetin improved the physicochemical properties and eating quality of yellow-feathered broiler chicken breast meat during 12 to 24 h postslaughter. Additionally, quercetin improved the postslaughter oxidative stress status and reduced protein and lipid oxidation levels. It also increased hydrogen bonding interactions and α-helix content during 6 to 12 h postslaughter and decreased β-sheet content during 12 to 24 h postslaughter in chicken breast MP. This resulted in improved postslaughter MP structure and thermal stability. The correlation results indicated that the enhancement of antioxidant capacity and MP structure enhanced the physicochemical and edible qualities of yellow-feathered broiler chicken breast meat. In conclusion, the current findings suggest that dietary supplementation with quercetin is an ideal approach for improving the eating quality of chicken meat, thereby broadening our understanding of theoretical and technological applications for improving the quality of chicken.

Loading rutin on surfaces by the layer-by-layer assembly technique to improve the oxidation resistance and osteogenesis of titanium implants in osteoporotic rats

The purpose of this study was to construct a rutin-controlled release system on the surface of Ti substrates and investigate its effects on osteogenesis and osseointegration on the surface of implants. The base layer, polyethylenimine (PEI), was immobilised on a titanium substrate. Then, hyaluronic acid (HA)/chitosan (CS)-rutin (RT) multilayer films were assembled on the PEI using layer-by-layer (LBL) assembly technology. We used scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy and contact angle measurements to examine all Ti samples. The drug release test of rutin was also carried out to detect the slow-release performance. The osteogenic abilities of the samples were evaluated by experiments on an osteoporosis rat model and MC3T3-E1 cells. The results (SEM, FTIR and contact angle measurements) all confirmed that the PEI substrate layer and HA/CS-RT multilayer film were effectively immobilised on titanium. The drug release test revealed that a rutin controlled release mechanism had been successfully established. Furthermore, the in vitro data revealed that osteoblasts on the coated titanium matrix had greater adhesion, proliferation, and differentiation capacity than the osteoblasts on the pure titanium surface. When MC3T3-E1 cells were exposed to H2O2-induced oxidative stress in vitro, cell-based tests revealed great tolerance and increased osteogenic potential on HA/CS-RT substrates. We also found that the HA/CS-RT coating significantly increased the new bone mass around the implant. The LBL-deposited HA/CS-RT multilayer coating on the titanium base surface established an excellent rutin-controlled release system, which significantly improved osseointegration and promoted osteogenesis under oxidative stress conditions, suggesting a new implant therapy strategy for patients with osteoporosis.

Evaluation of Oxidative Stress and Antioxidant Biomarkers in Chronic Cigarette Smokers: A Pilot Study.

Introduction The present study was undertaken to assess the status of oxidative stress in chronic cigarette smokers. Materials and methods Thirty adult male chronic cigarette smokers and an equal number of age and sex-matched normal subjects from the Deoghar district of Jharkhand state, India, were included in the study. The status of lipid peroxidation was determined using malondialdehyde (MDA), and the activities of enzymic antioxidants, such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), were determined using standard protocols. Results Results showed that the serum MDA levels were significantly increased, and the enzymic antioxidants were markedly decreased in chronic cigarette smokers compared to the normal subjects. Conclusion Our study demonstrated that oxidative stress is more pronounced in cigarette smokers compared to non-smokers. The number of cigarettes smoked plays a crucial role in increasing the reactive oxygen species and decreasing the cellular antioxidants.

Bone protective effects of the polysaccharides from Grifola frondosa on ovariectomy-induced osteoporosis in mice via inhibiting PINK1/Parkin signaling, oxidative stress and inflammation

BackgroundPolysaccharides from Grifola frondosa(GFP) have gained worldwide attention owing to their promising biological activities and potential health benefits. PurposeThis study aimed to investigate the effects of GFP on alleviation of osteoporosis in ovariectomized (OVX) mice and examine the underlying mechanism. MethodA mouse model of postmenopausal osteoporosis was established by OVX method, Forty eight C57BL/6 female mice were randomly divided into Normal group, OVX alone (Model group, n = 8), OVX + 10 mg/kg GFP (GFP-L group, n = 8), OVX + 20 mg/kg GFP (GFP-M group, n = 8), OVX + 40 mg/kg GFP (GFP-H group, n = 8), OVX + 10 mg/kg Estradiol valerate (Positive group, n = 8). ResultsThe results showed that compared with Model group, the concentrations of interleukin (IL)-1β, interleukin (IL)-6 and Tumor necrosis factor-α (TNF-α) were significantly reduced, the activity of superoxide dismutase (SOD) and glutathione (GSH) were significantly increased, the content of myeloperoxidase (MPO) and malondialdehyde (MDA) were significantly reduced, and the proteins levels of PINK1, Parkin, Beclin-1 and LC3-II were significantly decreased in the GFP groups. ConclusionThis study demonstrates that GFP alleviates ovariectomy-induced osteoporosis via reduced secretion of inflammatory cytokines, improvement in the oxidative stress status in the body, and inhibition of the PINK1/Parkin signaling pathway.

Effect of excitation wavelength and silver concentration on the biosynthesized GeO2 nanocrystals

The synthesis of GeO2 NCs and AgNPs/GeO2 NCs is carried out using germanium as starting material and pseudomonas putida in the presence of a low magnetic field for sustainable bacterial growth under oxidation stress and ambient conditions. The galvanic reaction has favored producing Ag NPs on the surface of GeO2 NCs without Ag2O formation. The Ag nanoparticles on the surface of GeO2 NCs will tune their optical properties. AgNPs/GeO2NCs exhibited enhanced PL and Raman intensities with peak shifts due to surface plasmon resonance. Bacterial inhibition will influence the formation of GeO2 nanostructures under oxidation stress. The generated reactive oxygen species during synthesis will form defects in the nanostructures. The defects, excitation wavelength, and Ag concentration will influence the optical properties of GeO2 nanostructures. The improvement in the bacterial culture, temperature regulation, and bacteria–germanium interactions can assist to synthesize optimum GeO2 nanostructures for optoelectronic applications.

Disruption of NADPH homeostasis by total flavonoids from Adinandra nitida Merr. ex Li leaves triggers ROS-dependent p53 activation leading to apoptosis in non-small cell lung cancer cells

Ethnopharmacological relevanceAdinandra nitida Merr. ex Li leaves serve as a herbal tea and hold a significant role in traditional Chinese medicine, being applied to assist in tumor treatment. Flavonoids present the primary bioactive constituents in Adinandra nitida Merr. ex Li leaves. Aim of the studyTo explore the potential of total flavonoids from Adinandra nitida Merr. ex Li Leaves (TFAN) in inhibiting non-small cell lung cancer (NSCLC) and further elucidate the underlying mechanisms. Materials and methodsHuman NSCLC cell lines and normal lung cell line were employed to assess the impact of TFAN (0–160 μg/mL for 24, 28 and 72 h) on cell proliferation in vitro. Immunofluorescence (IF) staining gauged p53 expression changes in NSCLC cells under TFAN present condition (150 μg/mL for 24 h). In vivo study utilized NSCLC cell derived xenograft tumors in nude mice, administering TFAN orally (200 and 400 mg/kg) for 14 days. Immunohistochemistry assessed Cleaved Caspase 3 expression change in A549 xenograft tumors treated with TFAN (400 mg/kg for 14 days). RNA-seq and KEGG analysis identified gene expression changes and enriched processes in A549 xenograft tumors treated with TFAN. CM-H2DCFDA and metabolomics assessed ROS level and GSH/GSSG pool changes in A549 cells under TFAN present condition. Cell viability assay and IF staining assessed A549 cell proliferation and p53 expression changes under H2O2-induced oxidative stress (0–40 μM for 24 h) and TFAN present conditions. GSEA and N-Acetyl-L-cysteine (NAC) rescue (0–1 μM for 24 h) analyzed the impact of TFAN on GSH de novo synthesis. NADPH/NADP+ pool measurement and NADPH rescue (0–10 μM for 24 h) analyzed the impact of TFAN on GSH salvage synthesis. GC-FID and HPLC-MS were utilized to detect ethanol and ethyl acetate residues, and to characterize the chemical constituents in TFAN, respectively. The total flavonoid content of TFAN was determined using a 330 nm wavelength. ResultsTFAN significantly inhibited A549 cells (wild-type p53) but not NCI–H1299 cells (p53-deficient), NCI–H596 cells (p53-mutant) or BEAS-2B in vitro. IF staining validated p53 genotype for the cell lines and revealed an increase in p53 expression in A549 cells after TFAN treatment. In vivo, TFAN selectively inhibited A549 xenograft tumor growth without discernible toxicity, inducing apoptosis evidenced by Cleaved Caspase 3 upregulation. RNA-seq and KEGG analysis suggested ROS biosynthesis was involved in TFAN-induced p53 activation in A549 cells. Elevated ROS level in TFAN-treated A549 cells were observed. Moreover, TFAN sensitized A549 cells to H2O2-induced oxidative stress, with higher p53 expression. Additionally, A549 cells compensated with GSH de novo synthesis under TFAN present condition, confirmed by GSEA and NAC rescue experiment. TFAN disrupted NADPH homeostasis to impair GSH salvage biosynthesis, supported by NADPH/NADP+ change and NADPH rescue experiment. The chemical constituents of TFAN, with acceptable limits for ethanol and ethyl acetate residues and a total flavonoid content of 68.87%, included Catechin, Epicatechin, Quercitroside, Camellianin A, and Apigenin. ConclusionThe disruption of NADPH homeostasis by TFAN triggers ROS-dependent p53 activation that leads to apoptotic cell death, ultimately suppressing NSCLC growth. These findings offer potential therapeutic implications of Adinandra nitida Merr. ex Li leaves in combating NSCLC.

Potential use of antioxidants for the treatment of chronic inflammatory diseases.

The excessive production of various reactive oxidant species over endogenous antioxidant defense mechanisms leads to the development of a state of oxidative stress, with serious biological consequences. The consequences of oxidative stress depend on the balance between the generation of reactive oxidant species and the antioxidant defense and include oxidative damage of biomolecules, disruption of signal transduction, mutation, and cell apoptosis. Accumulating evidence suggests that oxidative stress is involved in the physiopathology of various debilitating illnesses associated with chronic inflammation, including cardiovascular diseases, diabetes, cancer, or neurodegenerative processes, that need continuous pharmacological treatment. Oxidative stress and chronic inflammation are tightly linked pathophysiological processes, one of which can be simply promoted by another. Although, many antioxidant trials have been unsuccessful (some of the trials showed either no effect or even harmful effects) in human patients as a preventive or curative measure, targeting oxidative stress remains an interesting therapeutic approach for the development of new agents to design novel anti-inflammatory drugs with a reliable safety profile. In this regard, several natural antioxidant compounds were explored as potential therapeutic options for the treatment of chronic inflammatory diseases. Several metalloenzymes, such as superoxide dismutase, catalase, and glutathione peroxidase, are among the essential enzymes that maintain the low nanomolar physiological concentrations of superoxide (O2•-) and hydrogen peroxide (H2O2), the major redox signaling molecules, and thus play important roles in the alteration of the redox homeostasis. These enzymes have become a striking source of motivation to design catalytic drugs to enhance the action of these enzymes under pathological conditions related to chronic inflammation. This review is focused on several major representatives of natural and synthetic antioxidants as potential drug candidates for the treatment of chronic inflammatory diseases.