Lipid Peroxidation Products Research Articles

Abstract 4134449: Increasing Histidyl dipeptides in Skeletal Muscle Preserves Muscle Mass and Function During Heart Failure.

Background: In heart failure patients muscle wasting is a serious comorbidity. Oxidative stress and inflammatory cytokines, which cause muscle wasting, are increased in the skeletal muscle of heart failure patients. Oxidative stress leads to oxidation of lipids, generating toxic lipid peroxidation products, such as acrolein, which activates TNFa. Whether lipid peroxidation products accumulate in muscle and their removal from muscle influences muscle atrophy during heart failure has not been studied. In the muscle there are histidyl dipeptides, such as carnosine, ranging between 5-10 mM, synthesized via enzyme carnosine synthase (CARNS) that bind lipid peroxidation products. We examined whether intramuscular histidyl dipeptides influences heart failure-induced muscle wasting. Hypothesis: Histidyl dipeptides, alleviates intramuscular inflammation, preserves muscle mass and function during heart failure. Methods: Wild type (WT) and muscle-specific CARNS transgenic (Tg) mice were subjected to sham and transverse aortic constriction (TAC) surgeries. Cardiac function, muscle strength, atrophic and inflammatory markers, aldehyde modified protein, CARNS, and histidyl dipeptides were measured after 12 weeks. Results: In heart failure mice, weight of gastrocnemius (sham: 18.64 ± 1.73 vs TAC: 14.10 ± 0.90 mg, p=0.005), soleus (sham: 6.06 ± 0.82 vs TAC: 4.14 ± 0.70 mg, p=0.001), and tibialis anterior (sham:10.76 ± 1.62 vs TAC: 6.22 ± 1.83 mg, p=0.003), were decreased. Muscle strength was diminished (sham: 2.81 ± 0.36 vs TAC: 1.52 ± 0.27 N, p=0.004). Atrophic marker, atrogin1, aldehyde protein adducts and TNF-a were increased in the gastrocnemius muscle of TAC mice (~2-3-fold vs sham, p<0.05). CARNS expression and carnosine was decreased in different muscles of TAC mice (gastrocnemius; sham: 43.38 ± 20.46 vs TAC: 24.48 ± 6.49 nmoles/mg protein, p=0.03; soleus; sham: 22.04 ± 11.36 vs TAC: 11.02 ± 4.69 nmoles/mg protein, p=0.03; and tibialis; sham: 27.9 ± 11.70 vs TAC: 15.50 ± 3.48 nmoles/mg protein, p=0.01). Gastrocnemius muscle weight (WT TAC: 14.38 ± 1.08 vs Tg TAC: 17.30 ± 1.32 mg, p=0.004) and muscle strength (WT TAC: 3.06 ± 0.32 vs Tg TAC: 3.98 ± 0.41 N) were preserved, atrogin and TNFa expressions were decreased in gastrocnemius muscle of Tg TAC mice (~2-3-fold, p=0.04). Conclusion: Increasing intramuscular histidyl dipeptides alleviates muscle inflammation, preserves muscle mass, and function during heart failure.

Raloxifene Protects Oxygen-Glucose-Deprived Astrocyte Cells Used to Mimic Hypoxic-Ischemic Brain Injury

Perinatal asphyxia (PA) is a clinical condition characterized by oxygen supply suspension before, during, or immediately after birth, and it is an important risk factor for neurodevelopmental damage. Its estimated 1/1000 live births incidence in developed countries rises to 5–10-fold in developing countries. Schizophrenia, cerebral palsy, mental retardation, epilepsy, blindness, and others are among the highly disabling chronic pathologies associated with PA. However, so far, there is no effective therapy to neutralize or reduce PA-induced harm. Selective regulators of estrogen activity in tissues and selective estrogen receptor modulators like raloxifene have shown neuroprotective activity in different pathological scenarios. Their effect on PA is yet unknown. The purpose of this paper is to examine whether raloxifene showed neuroprotection in an oxygen–glucose deprivation/reoxygenation astrocyte cell model. To study this issue, T98G cells in culture were treated with a glucose-free DMEM medium and incubated at 37 °C in a hypoxia chamber with 1% O2 for 3, 6, 12, and 24 h. Cultures were supplemented with raloxifene 10, and 100 nM during both glucose and oxygen deprivation and reoxygenation periods. Raloxifene 100 nM and 10 nM improved cell survival—65.34% and 70.56%, respectively, compared with the control cell groups. Mitochondrial membrane potential was preserved by 58.9% 10 nM raloxifene and 81.57% 100 nM raloxifene cotreatment. Raloxifene co-treatment reduced superoxide production by 72.72% and peroxide production by 57%. Mitochondrial mass was preserved by 47.4%, 75.5%, and 89% in T98G cells exposed to 6-h oxygen–glucose deprivation followed by 3, 6, and 9 h of reoxygenation, respectively. Therefore, raloxifene improved cell survival and mitochondrial membrane potential and reduced lipid peroxidation and reactive oxygen species (ROS) production, suggesting a direct effect on mitochondria. In this study, raloxifene protected oxygen–glucose-deprived astrocyte cells, used to mimic hypoxic–ischemic brain injury. Two examiners performed the qualitative assessment in a double-blind fashion.

Abstract 4132769: Carnosine Supplementation Improves Walking Ability of Peripheral Arterial Disease Patients

Introduction: Peripheral artery disease (PAD) is a significant cause of cardiovascular morbidity and mortality, characterized by atherosclerosis in the skeletal muscle. Currently no mechanism-based therapeutics are available for this diseased population. In the muscle, histidyl dipeptides, such as carnosine (ranging between 5-10 mM) possess the abilities to bind with toxic lipid peroxidation products, such as acrolein. Preclincial studies show carnosine is depleted in the ischemic leg, increasing intramuscular carnosine improves angiogenesis, and blood flow in the ischemic leg. In humans, carnosine levels are increased in the muscle with carnosine supplementation. However, little is known whether carnosine levels are affected in PAD patients and its effect on their walking ability. Hypothesis: Carnosine supplementation will improve walking ability of PAD patients. Methods: We recruited normal (males: n=48; females: n=52; age: 50 ±10 years) and PAD subjects (males: n=52; females: n=41; age: 67±8 years; ankle brachial index (ABI): 0.65±0.18 measured urinary histidyl dipeptides, histidyl dipeptide-aldehyde conjugates, and oxidative stress markers: N-Acetyl-S-(3-hydroxypropyl)-L-Cysteine (3-HPMA) and N-acetyl-S-(2-carboxyethyl)-L-cysteine (CEMA), both are acrolein metabolites. PAD subjects (n=7; n=5 females and n=2 males; ABI: 0.68±0.08, age: 63±8 years) were supplemented with carnosine 2 g/day for 3 months, measured distance covered in a six-minute walk test (6MWT), ABI, blood profile at baseline and after completion. Results: In PAD subjects urinary carnosine was decreased (normal: 20±29 vs PAD:11±14 nmoles/mg creatinine, p<0.024), carnosine aldehdye conjugates were increased (carnosine propanal: 1.46±1.27 vs normal 0.56±1.08 nmoles/mg creatinine, p<0.0001; and carnosine propanol: 4.66±2.80 vs normal 1.52±1.21 nmoles/mg creatinine, p<0.0001). Oxidative stress markers were increased in the PAD subjects (CEMA: 341±210 vs normal: 119±86 ng/mg creatinine, p<0.0004 and 3HPMA: 6.57±6.32 ng/mg protien vs normal: 5.49±5.28 ng/mg creatinine, p<0.001). Following carnosine supplementation, blood profile was unchanged. Distance covered in 6MWT increased by 166±204 feet (before: 577±129 vs 743±152 feet after, p<0.048) and there was an increasing trend in ABI (before: 0.68±0.08 vs 0.80±0.19; after, p<0.18). Conclusion: Urinary carnosine could serve as a biomarker to evaluate PAD pathology and carnosine supplementation may improve walking ability of PAD patients.

INDICATORS OF FREE RADICAL OXIDATION AND ANTIOXIDANT PROTECTION IN THE ORAL FLUID OF CHILDREN WITH CHRONIC CATARRHAL GINGIVITIS AND DIABETIS MELLITUS

Failure of antioxidant mechanisms leads to an increase in lipid peroxidation products in the body, resulting in a non-specific cascade of cellular membrane damage. Proteins play a crucial role in metabolic processes, but when lipid peroxidation intensifies, protein modifications occur, leading to fragmentation, denaturation, and loss of biological activity. This disruption impairs tissue regenerative processes. The objective of this study was to analyze indicators of free radical oxidation and antioxidant protection in the oral fluid of children with chronic catarrhal gingivitis and type I diabetes mellitus. Materials and methods. A total of 170 children aged 12 to 16 were observed, including 130 who were examined and treated at the Endocrinology Department of the Regional Municipal Non-Profit Institution “Chernivtsi Regional Pediatric Clinical Hospital” for type 1 diabetes mellitus. Among them, 74 children had a disease duration of less than 5 years, including 65 with chronic catarrhal gingivitis. Sixty five children had diabetes for more than 5 years, with 44 of them diagnosed to have chronic catarrhal gingivitis. The children with chronic catarrhal gingivitis were further divided into groups based on their level of glycemic control: 1 child with optimal glycemic control, 66 with suboptimal glycemic control, and 42 with glycemic control at a high risk to life. The control group included 40 practically healthy children: 22 of them had clinically healthy periodontal tissues, and 18 children were diagnosed to suffer from chronic catarrhal gingivitis. Results. The study performed found out that children with chronic catarrhal gingivitis present increased indicators of lipid peroxide oxidation (protein oxidative modification, diene conjugates, malondialdehyde) and decreased activity of the enzyme in the antioxidant protective system of the oral fluid (whole protein, НS-groups, ceruloplasmin, activity of supermutase and catalase) in comparison with indicators of children without dental pathology. The most considerable changes were found in patients with chronic catarrhal gingivitis and type 1 diabetes mellitus, especially when the disease lasted more than 5 years (р<0,05).

The interplay between ferroptosis and inflammation: therapeutic implications for cerebral ischemia-reperfusion

Stroke ranks as the second most significant contributor to mortality worldwide and is a major factor in disability. Ischemic strokes account for 71% of all stroke incidences globally. The foremost approach to treating ischemic stroke prioritizes quick reperfusion, involving methods such as intravenous thrombolysis and endovascular thrombectomy. These techniques can reduce disability but necessitate immediate intervention. After cerebral ischemia, inflammation rapidly arises in the vascular system, producing pro-inflammatory signals that activate immune cells, which in turn worsen neuronal injury. Following reperfusion, an overload of intracellular iron triggers the Fenton reaction, resulting in an excess of free radicals that cause lipid peroxidation and damage to cellular membranes, ultimately leading to ferroptosis. The relationship between inflammation and ferroptosis is increasingly recognized as vital in the process of cerebral ischemia-reperfusion (I/R). Inflammatory processes disturb iron balance and encourage lipid peroxidation (LPO) through neuroglial cells, while also reducing the activity of antioxidant systems, contributing to ferroptosis. Furthermore, the lipid peroxidation products generated during ferroptosis, along with damage-associated molecular patterns (DAMPs) released from ruptured cell membranes, can incite inflammation. Given the complex relationship between ferroptosis and inflammation, investigating their interaction in brain I/R is crucial for understanding disease development and creating innovative therapeutic options. Consequently, this article will provide a comprehensive introduction of the mechanisms linking ferroptosis and neuroinflammation, as well as evaluate potential treatment modalities, with the goal of presenting various insights for alleviating brain I/R injury and exploring new therapeutic avenues.

The role of ACSL4 in stroke: mechanisms and potential therapeutic target.

Stroke, as a neurological disorder with a poor overall prognosis, has long plagued the patients. Current stroke therapy lacks effective treatments. Ferroptosis has emerged as a prominent subject of discourse across various maladies in recent years. As an emerging therapeutic target, notwithstanding its initial identification in tumor cells associated with brain diseases, it has lately been recognized as a pivotal factor in the pathological progression of stroke. Acyl-CoA synthetase long-chain family member 4 (ACSL4) is a potential target and biomarker of catalytic unsaturated fatty acids mediating ferroptosis in stroke. Specifically, the upregulation of ACSL4 leads to heightened accumulation of lipid peroxidation products and reactive oxygen species (ROS), thereby exacerbating the progression of ferroptosis in neuronal cells. ACSL4 is present in various tissues and involved in multiple pathways of ferroptosis. At present, the pharmacological mechanisms of targeting ACSL4 to inhibit ferroptosis have been found in many drugs, but the molecular mechanisms of targeting ACSL4 are still in the exploratory stage. This paper introduces the physiopathological mechanism of ACSL4 and the current status of the research involved in ferroptosis crosstalk and epigenetics, and summarizes the application status of ACSL4 in modern pharmacology research, and discusses the potential application value of ACSL4 in the field of stroke.

Cryopreservation of yellowtail kingfish (Seriola lalandi) sperm: Effects on metabolic enzymes and sperm quality

This study aimed to determine the effects of cryopreservation on sperm functionality and assess post-thaw sperm quality in yellowtail kingfish (Seriola lalandi). Milt designated for cryopreservation was frozen in a solution containing Cortland® medium +1.3 M Me2SO + 0.2 M glucose +2 % BSA in a 1:3 ratio (milt:cryoprotectant). Fresh milt was used as the control. The samples were frozen in 0.5 mL straws using a programmable freezing system. After thawing, the samples were evaluated for motility using Computer-Assisted Sperm Analysis (CASA) with a phase-contrast optical microscope. Additionally, flow cytometry and a multimodal reader were employed to assess various sperm parameters, including plasma membrane integrity (SYBR-14/PI), mitochondrial membrane potential (∆ΨMMit: JC-1), DNA fragmentation (TUNEL), ATP content (CellTiter-Glo®), intracellular calcium levels (Fluo-4 AM), lipid peroxidation (LPO), intracellular superoxide (DHE/SYTOX) and glutathione (GSH/GSSG) production, activities of glutathione peroxidase (GPx), catalase (CAT) and 8-oxoguanine glycosylase (OGG1), and detection of 8-hydroxydeoxyguanosine (8-OHdG). In cryopreserved milt, significant differences in motility (64.14 ± 2.2 %), plasma membrane integrity (61.5 ± 2.7 %), mitochondrial membrane potential (55.4 ± 2.4 %), DNA fragmentation (6.7 ± 1.1 %), ATP content (7.5 ± 1.5 nmol/109 spermatozoa), and intracellular calcium levels (59.8 ± 2.5 nM/109 spermatozoa) were observed compared to fresh milt. Differences were also noted in LPO, O2−i, GPx, GSH/GSSG, CAT, 8-OHdG and OGG1 between the cryopreserved and control groups. Additionally, correlations between sperm quality parameters, oxidative stress markers, and enzyme activity in cryopreserved spermatozoa were determined. These results suggest that cryopreservation induces significant alterations derived from oxidative damage in the functionality of yellowtail kingfish spermatozoa, and thus further research is necessary to explore the need for using antioxidants in the cryopreservation process of sperm from this species.

Assessment of clinical and laboratory parameters of oral сavity in patients with orthodontic systems

equipment (braces) on the biochemical parameters of oral fluid. Materials and methods. The study involved patients aged 18 to 22 years who are undergoing orthodontic treatment with a bracket system. Thegroup of examined persons was 25 people. The control group included 15 practically healthy people without braces. All patients underwent dental status assessment (Green – Vermillion hygiene index – simplified IG-Y; PMA inflammation index), laboratory examination of oral fluid before and after controlled oral hygiene. The biochemical analysis included determination of the acid-base balance in the oral fluid using a pH meter, assessment of free radical oxidation by chemiluminogram parameters by biochemiluminescence and by levels of molecular products of lipid peroxidation (diene and triene conjugates, Schiff bases), activity of the enzyme alkaline phosphatase, concentrations of total protein and lactate. Results and discussions. In patients undergoing orthodontic treatment with a bracket system, an insufficient level of oral hygiene and inflammatory manifestations in the gums of moderate severity were revealed. The level of the acid-base balance of the oral fluid before controlled hygiene was equal to 6.8±0.19, that is, below the range of normal pH values in the oral cavity. After hygiene, the pH of the oral fluid returned to the norm of 7.1±0.2. In patients with braces, activation of free radical oxidation and an increase in lipoperoxidation parameters – triene conjugates and Schiff bases, increased activity of alkaline phosphatase, total protein and lactate levels took place.Cоnclusion. After hygienic treatment of the oral cavity, the biochemical parameters did not differ statistically significantly from the control data.

The protective effect of Auricularia auricula polysaccharides on cyclophosphamide-induced immunosuppression and intestinal injury: A fecal microbiota transplantation study

Auricularia auricula polysaccharides (AHP) have potential immunomodulatory activities. However, the relationship between polysaccharides, intestinal microbiota, and immune activity remains unclear. Therefore, to explore this relationship, we performed immunosuppressive donor, fecal microbiota transplantation (FMT), and pseudo-sterile mouse validation experiments. Our results showed that AHP enhanced immunomodulatory activity, as evidenced by the alleviation of cyclophosphamide (Cy)-induced body weight loss, spleen and thymus atrophy, and splenic and colonic lesions. Additionally, AHP effectively regulated antioxidant enzyme activities (SOD, T-AOC, CAT, GSH-Px), lipid peroxidation product content (MDA), levels of immune factors (TNF-α, IFN-γ, IL-2, IL-4, IL-6), immunoglobulins (sIgA), and the expression of tight junction proteins (Muc2, ZO-1, Occuldin, and Claudin-1) in the colon of immunosuppressed mice. FMT results demonstrated that FAHP displayed superior immunomodulatory ability and could further affect the abundance of Dehydroepiandrosterone, Prostaglandin E2, Cholesterol and Pregnenolone by increasing the abundance of unclassified_Muribaculaceae, Desulfovibrio, Alloprevotella, and unclassified_Oscillospiraceae and decreasing that of Staphylococcus, Bacteroides, and Corynebacterium. Pseudo-sterilization experiments showed that AHP was unable to increase antioxidant enzyme activity, reduce lipid peroxidation content, or promote the release of immune factors. In brief, AHP did not alleviate Cy-induced immunosuppression after microbiota elimination. In summary, AHP exerted a protective effect against Cy-induced immunosuppression and intestinal damage by modulating intestinal microbiota and its metabolites. These findings provide new insights into the mechanisms by which AHP, as a functional food or drug, promotes immune response regulation and maintains intestinal health.

Influenza and non-influenza ARVI in children. A relationship between cytokine profile, parameters of the “lipid peroxidation – antioxidant defense system” as well as clinical and laboratory indicators

Influenza and acute respiratory viral infections (ARVI) impose a substantial damage to the population health in the Russian Federation due to their seasonal circulation and predominantly affect young children. A very few data on the cytokine profile, the nonspecific LPO — AOD system and their relationships with clinical characteristics in such diseases in preschool children are available. The aim of this study was to assess the cytokine profile, LPO — AOD system parameters and their relationship with the clinical and laboratory characteristics of diseases in preschool children with influenza and other ARVI. 86 preschool children (3–6 years old) were examined: with an established diagnosis of influenza (n = 31), non-influenza ARVI (n = 28), apparently healthy children (control group (n = 27). All pediatric samples were analyzed by enzyme-linked immunosorbent assay assessing blood serum concentrations of C-reactive protein and cytokines IL-1β, IL-4, IL-6, IL-8, TNFα, IFNα, IFNγ. Spectrophotometric, fluorometric and enzyme immunoassay methods to assess the state of the “LPO — AOD” system were used. In the group of children with influenza vs other ARVI, a higher incidence of intoxication syndrome was revealed. Cytokine profile in children from both clinical groups compared with control cohort was featured with higher indicators of both pro-inflammatory and anti-inflammatory origin. Children with non-influenza ARVI, had increased magnitude of LPO final products in the nonspecific LPO — AOD system along with lowered concentration of fat-soluble vitamins, general antioxidant activity, GSH level, and SOD activity. In the group with influenza, the level of primary and final lipid peroxidation products was increased, whereas that of for retinol,α-tocopherol, and total antioxidant activity was decreased paralleled with higher GSSG and SOD levels. Numerous correlations were noted in the group of children with ARVI: IL-1β/ketones, IL-6/ketones, IL-8/ketones, TNFα/ketones, IL-4/ketones, IFNg/shortness of breath, IFNα/cough, double bonds/fever, double bonds/AST, SO/intoxication, retinol/fever, GSSG/cough. The influenza group differed in the following relationships: IL-4/ketones, IL-4/fever, IFNα/ketones, CDs/AST. It can be concluded that in preschool children with ARVI and influenza, changes in the cytokine profile are accompanied by increased pro- and anti-inflammatory cytokine levels, increased intensity of lipid peroxidation reactions along with reduced magnitude of antioxidant factors. In the group with ARVI, there was a relationship between the final toxic products of lipid peroxidation — Schiff bases — and the intoxication index, as well as the presence of protective mechanisms in the form of connections between interferons and disease clinical manifestations. The group with influenza was distinguished by the presence of protective relations, which may have a beneficial effect in the context of developing pathological process. The data obtained will help expand the understanding of the pathogenetic mechanisms related to immune reactivity and nonspecific lipid peroxidation reactions in preschool patients and formulate appropriate measures for correction.

FSP1 Acts in Parallel with GPX4 to Inhibit Ferroptosis in COPD.

Glutathione peroxidase 4 (GPX4) has recently been reported to play an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD). Ferroptosis suppressor protein-1 (FSP1) is a protein that defends against ferroptosis in parallel with GPX4, but its role in the pathogenesis of COPD remains unexplored, and further research is needed. Normal and COPD lung tissues were obtained from lobectomy and lung transplant specimens, respectively. FSP1-overexpressing mice were established by monthly transfection with AAV9-FSP1 through modified intranasal administration. The expression of FSP1, GPX4, and prostaglandin-endoperoxide synthase 2 (PTGS2) was measured by Western blotting, immunohistochemistry and other methods. The correlation between FSP1 and ferroptosis and the role of FSP1 in COPD were explored by screening the expression of ferroptosis-related genes in a COPD cell model after the inhibition and overexpression of FSP1. We then explored the underlying mechanism of low FSP1 expression in patients with COPD in vitro by methylated RNA immunoprecipitation (MeRIP)-qPCR. We found that cigarette smoke exposure can lead to an increase in lipid peroxide production and ultimately ferroptosis, which is negatively regulated by FSP1 activity. FSP1 overexpression can prevent ferroptosis and alleviate emphysema. Next, we found that decreased FSP1 expression was caused by increased m6A modification of FSP1 mRNA. Moreover, the level of FSP1 decreased in a YTHDF2-dependent manner. These results indicate that METTL3-induced FSP1 mRNA methylation leading to low FSP1 expression is a potential therapeutic target for COPD. This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Indices of carbohydrate metabolism and antioxidant system state during germination of aged wheat and triticale seeds treated with H(2)S donor

Hydrogen sulfide is a gasotransmitter molecule involved in the realization of many functions of the plant organism, including seed germination. Aging of seeds is shown to be accompanied by oxidative stress and reduced germination. The effect of exogenous hydrogen sulfide on the germination of aged cereal seeds has not been studied. The aim of the work was to estimate the effect of priming with NaHS as an H2S donor on wheat and triticale seeds previously subjected to natural aging. Seeds of winter wheat (Triticum aestivum) and winter ×Triticosecale were stored indoors for 4 years at fluctuating temperature and humidity. Aged seeds were treated with 0.2-5 mM NaHS solution for 3 h and germinated in Petri dishes for 3 days. The hydropriming­ treatment was used as a control. Amylase activity in grains, the biomass of shoots and roots, the content of total sugars, H2O2, lipid peroxidation products and anthocyanin, and the activity of antioxidant enzymes in seedlings were determined. It was shown that after the treatment with H2S donor, the activities of catalase and guaiacol peroxidase, as well as the content of anthocyanins were increased only in triticale seedlings. Nevertheless, treatment of seeds of both cereal species was followed by enhanced growth of shoots and roots, increase in amylase and superoxide dismutase activities, decrease in H2O2 and MDA contents, and elevated accumulation of sugars in shoots. It is concluded that the increase in germination of aged cereal seeds under the influence of H2S donor is caused by increased mobilization of reserve carbohydrates and modulation of antioxidant system activity. Such treatment can be considered as an effective tool to improve seedling growth. Keywords: amylase, antioxidant system, hydrogen sulfide, oxidative stress, seeds aging, total sugar, Triticosecale, Triticum aestivum

Upregulation of PRRX2 by silencing Marveld3 as a protective mechanism against radiation-induced ferroptosis in skin cells.

Radiation-induced skin injury (RISI) represents a significant complication in patients receiving radiotherapy and individuals exposed to nuclear accidents, characterized by a protracted wound-healing process relative to injuries from other etiologies. Current preventive and management approaches remain inadequate. Consequently, investigating efficacious intervention strategies that target the disease's progression characteristics holds significant practical importance. Small interfering RNA (siRNA) and overexpression plasmid were used to modulate the expression of Marvel domain containing 3 (Marveld3) and paired related homeobox 2 (PRRX2). Protein and mRNA levels were estimated by Western Blot and real-time PCR, respectively. Intracellular levels of Malondialdehyde (MDA), a terminal product of lipid peroxidation, were measured following the manufacturer's protocol for MDA assay kit. Similarly, intracellular levels of ferrous iron (Fe2+) and reactive oxygen species (ROS) were determined using their respective assay kits. Lipid peroxidation status within the cells was evaluated via BODIPY staining. Immunohistochemistry was conducted to ascertain the expression of PRRX2 in skin tissues collected at various time points following irradiation of rats. The H-score method was used to evaluate the percentage of positively stained cells and staining intensity. RNA sequencing, Gene Ontology (GO) analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were conducted by OE Biotech Company. In this study, our findings indicated that Marveld3 suppression could effectively inhibit lipid peroxidation levels in irradiated skin cells, concomitantly reducing intracellular Fe2+ content. Additionally, the silencing of Marveld3 effectively abrogated the impact of a ferroptosis agonist on cellular viability, resulting in the upregulation of 66 and 178 genes, as well as the downregulation of 188 and 31 genes in irradiated HaCaT and WS1 cells, respectively. Among the differentially expressed genes, the PRRX2 which was found to be involved in the process of ferroptosis, exhibited statistically significant upregulation. And the upregulation of PRRX2 expression may attenuate radiation-induced lipid peroxidation in skin cells, thereby functioning as a potential stress-responsive mechanism to counteract radiation effects. This study elucidates the role of Marveld3 in radiation-induced ferroptosis in skin cells. Inhibition of Marveld3 led to the upregulation of PRRX2, which subsequently resulted in a reduction of Fe2+ and ROS levels, as well as the suppression of lipid peroxidation. These effects collectively mitigated the occurrence of ferroptosis.

Metabolic pathways for removing reactive aldehydes are diminished in the skeletal muscle during heart failure.

Muscle wasting is a serious complication in heart failure patients. Oxidative stress and inflammation are implicated in the pathogenesis of muscle wasting. Oxidative stress leads to the formation of toxic lipid peroxidation products, such as 4-hydroxy-2-nonenal (HNE), which covalently bind with proteins and DNA and activate atrophic pathways. Whether the formation of lipid peroxidation products and metabolic pathways that remove these toxic products are affected during heart failure-associated skeletal muscle wasting has never been studied. Male C57BL/6J mice were subjected to sham and transverse aortic constriction (TAC) surgeries for 4, 8 or 14weeks. Different skeletal muscle beds were weighed, and the total cross-sectional area of the gastrocnemius muscle was measured via immunohistochemistry. Muscle function and muscle stiffness were measured by a grip strength meter and atomic force microscope, respectively. Atrophic and inflammatory marker levels were measured via qRT‒PCR. The levels of acrolein and HNE-protein adducts, aldehyde-removing enzymes, the histidyl dipeptide-synthesizing enzyme carnosine synthase (CARNS), and amino acid transporters in the gastrocnemius muscle were measured via Western blotting and qRT‒PCR. Histidyl dipeptides and histidyl dipeptide aldehyde conjugates in theGastrocnemius and soleus muscles were analyzed by LC/MS-MS. Body weight, gastrocnemius muscle and soleus muscle weights and the total cross-sectional area of the gastrocnemius musclewere decreased after 14weeks of TAC. Heart weight, cardiac function, grip strength and muscle stiffness were decreased in the TAC-operated mice. Expression of the atrophic and inflammatory markers Atrogin1 and TNF-α, respectively, was increased ~ 1.5-2fold in the gastrocnemius muscle after 14weeks of TAC (p < 0.05 and p = 0.004 vs sham). The formation of HNE and acrolein protein adducts was increased, and the expression of the aldehyde-removing enzyme aldehyde dehydrogenase (ALDH2) was decreased in the gastrocnemius muscle of TAC mice. Carnosine (sham: 5.76 ± 1.3 vs TAC: 4.72 ± 0.7nmol/mg tissue, p = 0.04) and total histidyl dipeptide levels (carnosine and anserine; sham: 11.97 ± 1.5 vs TAC: 10.13 ± 1.4nmol/mg tissue, p < 0.05) were decreased in the gastrocnemius muscle of TAC mice. Depletion of histidyl dipeptides diminished the aldehyde removal capacity of the atrophic gastrocnemius muscle. Furthermore, CARNS and TAUT protein expression were decreased in the atrophic gastrocnemius muscle. Our data reveals that reduced expression of ALDH2 and depletion of histidyl dipeptides in the gastrocnemius muscle during heart failure leads to the accumulation of toxic aldehydes and might contribute to muscle wasting.

Activity of photosynthetic pigments and the antioxidant system in sunflower under drought stress

Background. The study is of particular importance in view of the increasing pace of global climate aridization. The article highlights the results of an experiment on the effect of drought on the physiological status of seedlings of cv. ‘Poseidon 625’ representing an important food crop – sunflower (Helianthus annuus L.). Materials and methods. The experiment included a group of control samples grown with sufficient moisture and four impact groups subjected to osmotic stress. The intensity of accumulation of lipid peroxidation products (LPP) was measured by the reaction of malondialdehyde (MDA) with thiobarbeturic acid; catalase activity was assessed by a photocolorimetric method based on the interaction between hydrogen peroxide and potassium iodide; the content of pigments (Chl a, Chl b, Сar) was calculated spectrophotometrically in acetone extract. Results. The degree of POL accumulation in the impact groups of the experiment was found to be many times higher than the values in the control samples, which was confirmed by a rapid increase in the MDA concentration in response to a growing water shortage. The accumulation of oxygen free radicals triggered the mechanisms of antioxidant protection of seedlings by synthesizing catalase, the concentration of which increased proportionally to the accumulation of POL. At the same time, the rapid accumulation of POL in the absence of irrigation and under osmotic stress of 3 and 5 atm led to a suppression of low-molecular-weight components of protection (carotenoids) and activation of their synthesis only when critical values of osmotic stress were reached. Conclusion. As a result of the experiment, catalase was identified as the main component of antioxidant protection in sunflower seedlings. Due to the activation of its synthesis, the concentrations of Chl a and Chl b decreased, attesting to the activation of the mechanisms protecting the photosynthetic activity in seedlings, and their antioxidant status.

Dynamics of Free Radical Oxidative Processes During the Latent Period of Experimental Metastasizing to the Liver

Purpose — to investigate the dynamics of the content of antioxidant enzymes superoxide dismutase 1 (SOD1), glutathione peroxidase 1 (GPO1), glutathione reductase (GR) and lipid peroxidation products diene conjugates (DC), malondialdehyde (MDA) in the spleen and liver during the latent period of growth and metastasis of experimental tumor.Materials and methods. Using 28 white male rats, a model of hematogenous liver metastasis was created by transplanting sarcoma 45 cells (S45) into the spleen, previously lead out under the skin 3 weeks before. Previously, was determined that a tumor visualized in the spleen at 5 weeks, and liver metastases at 7 weeks after transplantation S45. Levels of SOD1, GPO1, GR and MDA were determined using ELISA and DC by biochemical method in spleen and liver homogenates during the latent period of tumor growth and metastasis (1–2 weeks post-transplantation).Results. Significant changes (1.5–5.2 times, р &lt; 0.050–0.001) in studied factors levels were observed compared to intact rats and rats with the spleen lead out. Activation of lipid peroxidation and antioxidant system was noted in the spleen (tumor-carrying organ) during tumor growth and metastasis. At the same time, in the liver (the target organ of metastasis) observed also increased lipid peroxidation but simultaneously a pronounced decreased GR levels (5 times, p &lt; 0.001) without affecting SOD1 levels.Conclusion. Liver tissue exhibited the inferiority of antioxidant protection and the formation of pro-oxidant condition during the latent period of tumor growth, which may prepare the soil for metastasis.

Level of prooxidant-antioxidant status in highly productive cows with comorbid obstetric, gynecological and orthopedic pathology

The lipid peroxidation and antioxidant defense system is a balanced system responsible for the processing and utilization of lipids in the body’s cells. It plays an important role in lipid metabolism, cell protection and overall health of the body. Its proper functioning is necessary to ensure optimal functioning of cells and organs throughout the body. The article clinically and experimentally substantiates the pathogenetic role of lipid peroxidation products and the level of antioxidant protection in cows with endometritis and purulent-necrotic processes in the finger area, as well as with the comorbid course of endometritis and orthopedic pathology. It has been shown that multimorbid manifestation is accompanied by a more severe course than individual diseases. It has been established that the development of postpartum endometritis and purulent-necrotic lesions of the limbs in cows is accompanied by a highly significant increase in lipid peroxidation products in the blood serum against the background of a decrease in the amount of antioxidant enzymes, with the exception of ceruloplasmin. Moreover, these changes are accompanied by a sharp jump in the comorbid course of endometritis and orthopedic pathology in highly productive animals.

Lipid peroxidation products induce carbonyl stress, mitochondrial dysfunction, and cellular senescence in human and murine cells.

Lipid enals are electrophilic products of lipid peroxidation that induce genotoxic and proteotoxic stress by covalent modification of DNA and proteins, respectively. As lipid enals accumulate to substantial amounts in visceral adipose during obesity and aging, we hypothesized that biogenic lipid enals may represent an endogenously generated, and therefore physiologically relevant, senescence inducers. To that end, we identified that 4-hydroxynonenal (4-HNE), 4-hydroxyhexenal (4-HHE) or 4-oxo-2-nonenal (4-ONE) initiate the cellular senescence program of IMR90 fibroblasts and murine adipose stem cells. In such cells, lipid enals induced accumulation of γH2AX foci, increased p53 signaling, enhanced expression of p21Cip1, and upregulated the expression and secretion of numerous cytokines, chemokines, and regulatory factors independently from NF-κB activation. Concomitantly, lipid enal treatment resulted in covalent modification of mitochondrial proteins, reduced mitochondrial spare respiratory capacity, altered nucleotide pools, and increased the phosphorylation of AMP kinase. Lipid-induced senescent cells upregulated BCL2L1 (Bcl-xL) and BCL2L2 (Bcl-w). and were resistant to apoptosis while pharmacologic inhibition of BAX/BAK macropores attenuated lipid-induced senescence. Insitu, the 4-HNE scavenger L-carnosine ameliorated the development of the cellular senescence, while in visceral fat of obese C57BL/6J mice, L-carnosine reduced the abundance of 4-HNE-modified proteins and blunted the expression of senescence biomarkers CDKN1A (p21Cip1), PLAUR, BCL2L1, and BCL2L2. Taken together, the results suggest that lipid enals are endogenous regulators of cellular senescence and that biogenic lipid-induced senescence (BLIS) may represent a mechanistic link between oxidative stress and age-dependent pathologies.

Deubiquitylase USP52 Promotes Bladder Cancer Progression by Modulating Ferroptosis through Stabilizing SLC7A11/xCT.

Bladder cancer (BLCA) is a prevalent cancer with high case-fatality rates and a substantial economic burden worldwide. Understanding its molecular underpinnings to guide clinical management is crucial. Ferroptosis, a recently described non-apoptotic form of cell death, is initiated by the lethal accumulation of iron-dependent lipid peroxidation products. Despite growing interest, the roles and vulnerabilities determining ferroptosis sensitivity in BLCA remain unclear. Re-analysis of single-cell RNA data reveals a decrease in high-ferroptosis cancer cells as BLCA advances. USP52/PAN2 is identified as a key regulator of ferroptosis in BLCA through an unbiased siRNA screen targeting 96 deubiquitylases (DUBs). Functionally, USP52 depletion impedes glutathione (GSH) synthesis by promoting xCT protein degradation, increasing lipid peroxidation and ferroptosis susceptibility, thus suppressing BLCA progression. Mechanistically, USP52 interacts with xCT and enzymatically cleaves the K48-conjugated ubiquitin chains at K4 and K12, enhancing its protein stability. Clinical BLCA samples demonstrate a positive correlation between USP52 and xCT expression, with high USP52 levels associated with aggressive disease progression and poor prognosis. In vivo, USP52 depletion combined with ferroptosis triggers imidazole ketone Erastin (IKE) synergistically restrains BLCA progression by inducing ferroptosis. These findings elucidate the role of the USP52-xCT axis in BLCA and highlight the therapeutic potential of targeting USP52 and ferroptosis inducers in BLCA.

The Role of Arginase and some Parameters in Diabetic Type 2 Patients with and without Retinopathy

Background: Oxidative stress plays a major role in the pathogenesis of diabetes mellitus by damaging cellular organelles and enzymes in blood such as arginase1, insulin and glutathione s-transferase; increasing lipid peroxidation such as malondialdehyde and increasing insulin resistance which can lead to diabetic complications such as diabetic retinopathy. Objectives: To explore the relationship of oxidative stress to the development of diabetic retinopathy by measuring the levels of Arginase1, the activity of glutathione s-transferase enzyme, and the levels of malondialdehyde as a secondary product of lipid peroxidation (biomarker for oxidative stress). Patients and MethodsThis study was conducted from November 2022 to January 2023 at the Ibn Al-Haitham Teaching Eye Hospital in Baghdad, the University of Baghdad / Department of Chemistry and the National Diabetes Centre for Treatment and Research at Al-Mustansyrriah University. This study was conducted on 120 subjects distributed as follows: 40 non-diabetic obese controls, 40 type 2 diabetics with no retinopathy, and 40 type 2 diabetic retinopathy patients, between 30 and 65 years of age. All groups were subjected to tests: measuring fasting blood glucose (FBG), HbA1c, lipid profile (cholesterol, triglycerides, HDL- cholesterol, LDL- cholesterol, and VLDL cholesterol), serum total arginase1, malondialdehyde glutathione s -transferase, body mass index (BMI), and waist-to-hip ratio. Results: Mean arginase1 levels were significantly higher in diabetic patients than in diabetic retinopathy and control groups (p ≤ 0.05). The mean xidative stress marker malondialdehyde concentration was significantly higher in diabetic retinopathy patients than in type 2 diabetics and the control group (P ≤ 0.05). The mean glutathione s-transferase activity in diabetic retinopathy patients was significantly higher than in the control group and type 2 diabetics (P ≤ 0.05). Conclusion: There is a relationship between oxidative stress and the development of diabetic retinopathy, where the levels of arginase1 and malondialdehyde increased and the activity of glutathione s –transferase enzyme increased as a result of oxidative stress and inflammation associated with complications of type 2 diabetes.