Superoxide Dismutase Research Articles

Preclinical antidiabetic and antioxidant effects of Erythrophleum africanum (benth.) harms in streptozotocin-induced diabetic nephropathy.

This study investigated the antidiabetic effects of the methanolic extract of E.africanum (MEEA) stem bark on streptozotocin (STZ)-induced diabetic nephropathy (DN) in Wistar rats. The invitro enzyme (α-amylase) inhibitory activity of MEEA was measured using a standard procedure. Diabetic rats with fasting blood glucose above 250 mg/dL were considered diabetic and were divided intothe following groups: control (distilled water-treated), diabetic-control, diabetic metformin (100 mg/kg), diabetes + MEEA (150 mg/kg), and diabetes + MEEA (300 mg/kg) via oral gavage once daily for 14days. At the end of the experimental period, kidney tissues were collected for biochemical and histological analyses. Kidney apoptosis and marker gene expression were measured by real-time quantitative PCR. MEEA exhibited α-amylase inhibitory effects. MEEA significantly (p<0.05) reduced the STZ-induced increases in blood glucose, serum urea, serum creatinine, uric acid, alanine aminotransferase, alkaline phosphatase, and malondialdehyde and increased the STZ-induced decreases in superoxide dismutase, catalase, and reduced glutathione. In addition, MEEA protects against DN by significantly downregulating the mRNA expression of cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), cAMP-response binding protein (CREB), and cFOS and upregulating B-cell lymphoma 2 (Bcl-2), suggesting that the nephroprotective ability of MEEA is due to the modulation of the cAMP/PKA/CREB/cFOS signaling pathway. Furthermore, MEEA treatment protected against histopathological alterations observed in diabetic rats. The data from this study suggest that MEEA modulates glucose homeostasis and inhibits redox imbalance in DN rats.

P-002 Acetate attenuates cyclophosphamide-induced testicular toxicity by modulating steroidogenic and apoptotic signaling via an oxidative stress-dependent pathway

Abstract Study question Will acetate therapy attenuate cyclophosphamide-induced testicular toxicity when used concomitantly? Summary answer Acetate attenuated cyclophosphamide-induced testicular toxicity by modulating steroidogenic and apoptotic signaling via an oxidative stress-dependent pathway. What is known already Cyclophosphamide is an anthracycline that is effective in the management of cancers. Nonetheless, a major drawback of this drug is its testicular toxicity. Available evidence in the literature revealed that cyclophosphamide induces testicular toxicity by targeting steroidogenesis and activating oxidative stress, inflammation, and apoptosis. On the other hand, acetate has been reported to exert antioxidant, anti-inflammatory, and anti-apoptotic activities. However, there is no report on the effect of acetate on cyclophosphamide-induced testicular toxicity. Study design, size, duration This is a prospective experimental study using animal model. Twenty-four sexually mature litter-mate male Wistar rats of comparable weight were used for the study. The study lasted 2 weeks. Participants/materials, setting, methods Animals were acclimatized for two weeks, and then randomized into four groups. The control rats received 0.5mL of distilled water p.o for 14 days, acetate-treated rats received 200 mg/kg/day of acetate p.o for 14 days, cyclophosphamide-treated rats received 150 mg/kg of cyclophosphamide i.p on day 8, while acetate + cyclophosphamide-treated rats received treatment as acetate-treated and cyclophosphamide-treated. The doses of drugs used were the Human Equivalent doses for rats and as earlier reported. Main results and the role of chance Acetate attenuated cyclophosphamide-induced alterations in testicular histoarchitecture and the rise in testicular activities of gamma glutamyl transferase and lactate dehydrogenase activities, and lactate levels. In addition, acetate abrogated cyclophosphamide-induced rise in testicular malondialdehyde, TNF-α, IL-1β, NF-kB, and Bax levels, and myeloperoxidase, caspase 9, and caspase 3 activities, which was accompanied by acetate-driven blockade of cyclophosphamide-induced decline in reduced glutathione, Nrf2, and Bcl-2 levels, and glutathione peroxidase, glutathione-S-transferase, superoxide dismutase and catalase activities. Furthermore, acetate alleviated cyclophosphamide-induced suppression of circulatory levels of FSH, LH, and testosterone, testicular concentrations of 3β-HSD, 17β-HSD, and testosterone, and spermatogenesis and lowered sperm quality. Limitations, reasons for caution This was an experimental study using a rat model; hence, the results of this study should be extrapolated to humans with caution. Clinical trials are recommended to validate these findings. Wider implications of the findings The present study revealed the protective effect of acetate treatment on cyclophosphamide-induced testicular toxicity. These findings provide convincing evidence of the attenuating effect of acetate in cyclophosphamide-induced testicular toxicity. Trial registration number N/A

P-104 Cytoprotective effect of melatonin in prepubertal Sertoli cells: role in cadmium-induced idiopathic male infertility

Abstract Study question Melatonin (MLT) is a cytoprotective agent holding potential to prevent cadmium (Cd) toxicity and its impact in testicular function and fertility. Summary answer MLT produced a potent cytoprotective effect in Sertoli cells (SCs) exposed to Cd-toxicity, reducing its H2O2 generation and reductive stress effects by modulation Nrf2 activity. What is known already MLT a hormonal substance exhibiting strong antioxidative and antiapoptotic properties, has been proved to hold great potential as cytoprotec tive agent in a range of tissues and cell models exposed to Cd toxicity, including the rat bone, ovaries, human MSC, and even mammalian testes. However, cellular and mechanistic aspects of this effect of MLT in testes remain poorly understood. Cd is a pro-oxidant heavy metal reported to interfere with testicular function and fertility. Both the toxicity of Cd and the cytoprotective function of MLT involve redox-dependent mechanisms with the activity of Nrf2 transcription factor as key player on the cellular stress response. Study design, size, duration Porcine pre-pubertal SCs were maintained at 37 °C in a 5% CO2 humidified atmosphere in the absence (unexposed-control group) or presence of 5 or 10µM CdCl2 (Sigma Chemical Co.) and/or 50nM MLT (Sigma-Aldrich) for 48h in HAMF12 (Euroclone) supplemented with 0.166nM retinoic acid (Sigma-Aldrich Co.) and 5/500ml of Insulin-Transferrin-Selenium (ITS) + Premix (Cat. No. 354352; Corning). Participants/materials, setting, methods After treatment the parameters investigated in the study were: a) SCs viability (MTT test); b) SCs functionality parameters AMH and inhibin B. (real-time PCR and ELISA analysis); c) Intracellular and extracellular ROS production ( with Amplex Red and DCFH-DA fluorescent probe respectively); d) Catalase enzyme activity (spectrophotometric assay procedure); e) Nrf2 and other transcription factors (by semi-quantitative microplate confocal microscopy analysis); f) Intracellular and extracellular levels of GSH and other thiols (by HPLC); g) c-Jun and ERK1/2 (Immunoblot). Main results and the role of chance MLT co-treatment significantly prevented the effects of Cd on cell viability and abnormal ROS production. The reduction of H2O2 efflux was very important and combined with an increased CAT activity that was demonstrated upon MLT treatment significantly in SCs exposed to 5μMCd. The co-treatment with MLT compared to Cd treatment, showed a trend toward a significant recovery of AMH and inhibin B gene expression and secretion levels, respectively, demonstrating the cytoprotective effect of MLT in SCs exposed to Cd toxicity at the functional level. In Cd-treated cells, MLT significantly reduced the expression and nuclear translocation of Nrf2 protein, and decreased GSTP induction. However, MLT reduced Cys uptake and stimulated GSH efflux when the cells were treated with 10 µM Cd, suggesting a specific role for the Nrf2-GSTP axis in the cytoprotective response to MLT of Cd-treated SCs. Finally MLT induced MAPK-ERK1-2 pathway and c-Jun transcription factor when the cells were treated with Cd. MLT is a cytoprotective agent in SCs exposed to Cd toxicity capable to restore cell viability and function. Its activity is associated with the modulation of Nrf2 and other stress response genes important in the control of thiols and H2O2 metabolism of SCs, including cJun and MAPKs. Limitations, reasons for caution Other ROS-scavenging genes other than CAT may have a role in the antioxidant response to MLT of SCs, such as superoxide dismutase, glutathione peroxidases, and peroxyredoxins that are worth investigating further. Wider implications of the findings The aspects that emerged from our study and, described for the first time in these cells, warrant further preclinical and clinical investigation holding potential in the treatment of Cd-induced testicular damage and fertility problems. Trial registration number Not applicable

P-100 Prolonged exposure to polyethylene glycol (peg) 6000 severely affects the reproductive system of male Wistar rats

Abstract Study question Studies have shown that microplastics have adverse effects on aquatic reproductive health. Is there any potential effect of Polyethylene glycol (PEG)6000 on rodents reproductive system? Summary answer Despite its wide use in industry, medicine and cosmetic formulations, prolonged exposure to PEG 6000 may pose great risk, especially causing reproductive toxicity. What is known already The usage of Polyethylene glycol (PEG) 6000, a microplastic, is rising in industries, medicine, and cosmetic formulations. They serve as ointment bases, capsules, liquid prescription, suppositories and recently used to alleviate constipation. More so, it is one of the pivotal ingredients in soaps, detergents, and also used as chemical intermediates in textile, leather and plastic industries. Additionally, PEG 6000 is widely used in drug delivery systems and tissue engineering. Although approved for internal consumption by regulatory agencies like FDA, there are evidences of its developmental toxicity. Study design, size, duration Fifteen(15) rats weighing between 200-250g were used in this study. The rats were divided into three groups(A, B and C), of 5 rats in each group. Group A was designated as the control group while groups B and C were the test groups. Group A received distilled water, while groups B and C were administered 50 mg/kg and 200 mg/kg body weight of polyethylene glycol PEG6000 respectively via oral gavage daily for 6 weeks. Participants/materials, setting, methods After 6 weeks, the rats were sacrificed as reproductive organs and serum obtained. Enzyme immunosorbent assay (ELISA) was used for hormonal assays. Reproductive organs were weighed and tissue damage was determined using Haemoxylin /eosin staining. Antioxidant enzymes levels (Catalase, Superoxide dismutase), Glutathione peroxidise) and Lipid peroxidation (Malondialdehyde) were also evaluated. Computer-assisted sperm assay (CASA) was used to evaluate sperm parameters and sperm movement kinetics. Main results and the role of chance PEG 6000 caused a reduction in the weights of the rats in groups B and C when compared with control group. There was no significant change in the relative testicular and seminal vesicle organ/body weight ratio in groups B and C, however a pattern of reduction was seen in the organ/body weight ratio of epididymis and prostate, when compared to control group. Serum hormonal levels of Follicle stimulating hormone and Luteinizing hormone were reduced in the test groups when compared to control, but not significant. Testosterone was significantly reduced in groups B and C, while estradiol levels were significantly elevated in groups B and C when compared to control group. There was significant reduction in the levels of Superoxide dismutase, Catalase and Glutathione peroxidase, and a significant increase in Malondialdehyde levels in groups B and C when compared to the control group. Histopathological changes were also seen in the testes, epididymis, seminal vesicles and prostate glands of rats in the test groups. Semen analyses showed statistically significant reductions (p &amp;lt; 0.05) in sperm concentration, motility, vitality and sperm movement kinetics of rats in the test groups when compared to the control group. Limitations, reasons for caution Although this study was done in rats, it may not be easy to extrapolate the present findings to humans. However, this calls for caution as primates and rodents are both members of the superorder Euarchontoglires. Wider implications of the findings Prolonged ingestion of PEG 6000 affects male reproductive system, alters hormonal levels, reduces sperm parameters and induces oxidative stress. Interestingly, the disruptive effects of PEG 6000 are dose-dependent, and may lead to male infertility. Trial registration number not applicable

Mitigation of Abiotic Stresses in Plants through Nutrient Management

The food demand over the world is increasing due to the rapid increase in the population. Direct and indirect effects of climate change have severely affected the growth and development of crops. Of these, abiotic stress factors are reported to cause a reduction in crop productivity ranging from 51 percent to 82 percent. Abiotic stresses like drought, waterlogging stress, salt stress, soil acidity, metal toxicities and temperature variations have overwhelming impact on the growth and productivity of crops. Abiotic stress causes increase in reactive oxygen species (ROS) levels and affects various physiological processes, causing reduction in plant growth and yield. Nutrient management proves to be an effective strategy for alleviating various abiotic stress factors affecting agricultural crops. Nutrients such as nitrogen, potassium, calcium and magnesium increase the production of antioxidant enzymes such as superoxide dismutase, peroxidase, catalase and reduces ROS production. Micronutrients such as iron, boron and zinc as well as biofertilizers improve plant adaptation to various stresses through activation of antioxidant enzymes. Current review focuses on the impact of mineral nutrients, organic amendments and biofertilizers in alleviating abiotic stress in agricultural crops.

Hematological and neurological impact studies on the exposure to naturally occurring radioactive materials

Naturally Occurring Radioactive Materials (NORM) contribute to everyone's natural background radiation dose. The technologically advanced activities of the gas and oil sectors produce considerable amounts of radioactive materials as industrial by-products or waste products. The goal of the current study is to estimate the danger of long-term liability to Technologically Enhanced Naturally Occurring Radioactive Materials (TE-NORM) on blood indices, neurotransmitters, oxidative stress markers, and β-amyloid in the cerebral cortex of rats' brains. Twenty adult male albino rats were divided into two equal groups (n = 10): control and irradiated. Irradiated rats were exposed to a total dose of 0.016 Gy of TE-NORM as a whole-body chronic exposure over a period of two months. It should be ''The results showed no significant changes in RBC count, Hb concentration, hematocrit percentage (HCT%), and Mean Corpuscular Hemoglobin Concentration (MCHC). However, there was a significant increase in the Mean Corpuscular Volume of RBCs (MCV) and a significant decrease in cell distribution width (RDW%) compared to the control. Alteration in neurotransmitters is noticeable by a significant increase in glutamic acid and significant decreases in serotonin and dopamine. Increased lipid peroxidation, decreased glutathione content, superoxide dismutase, catalase, and glutathione peroxidase activities indicating oxidative stress were accompanied by increased β-amyloid in the cerebral cortex of rats' brains. The findings of the present study showed that chronic radiation liability has some harmful effects, that may predict the risks of future health problems in occupational radiation exposure in the oil industries. Therefore, the control of exposure and application of sample dosimetry is recommended for health and safety.

Protective Role of Ellagic Acid Against Ethanol-Induced Neurodevelopmental Disorders in Newborn Male Rats: Insights into Maintenance of Mitochondrial Function and Inhibition of Oxidative Stress.

Ellagic acid (EA) exerts, neuroprotective, mitoprotective, anti-oxidative and anti-inflammatory effects. We evaluated protective effect of EA on ethanol-induced fetal alcohol spectrum disorders (FASD). A total of 35 newborn male rats were used, divided into five groups, including; control (normal saline), ethanol (5.25 g/kg per day), ethanol (5.25 g/kg per day) + EA (10 mg/kg), ethanol (5.25 g/kg per day) + EA (20 mg/kg) and ethanol (5.25 g/kg per day) + EA (40 mg/kg). Thirty-six days after birth behavioral tests (Morris water maze and Elevated Plus Maze), tumor necrosis factor-α (TNF-α) levels, oxidative markers (malondialdehyde, glutathione and superoxide dismutase), mitochondrial examination such as succinate dehydrogenases (SDH) activity, mitochondrial swelling, mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) formation were analyzed. The results revealed that ethanol exposure adversely affected cognitive and mitochondrial functions and as well as induced oxidative stress and inflammation in brain tissue. However, EA (20 and 40 mg/kg) administration effectively prevented the toxic effects of ethanol in FASD model. These findings demonstrate that ethanol application significantly impairs the brain development via mitochondrial dysfunction and induction of oxidative stress. These data indicate that EA might be a useful compound for prevention of alcohol-induced FASD.

Characterization of Physiological and Biochemical Attributes of Neem (Azadirachta indica A. Juss) under Salinity Stress

Salinity poses a significant threat to agricultural productivity worldwide, with its detrimental effects on plant growth and physiological processes. Understanding the mechanisms by which plants respond to salt stress is crucial for developing strategies to mitigate its impact on crop yield and sustainability. To address this issue, a pot study was conducted to determine the effect of salt stress on the physiological and biochemical attributes of neem (Azdiarchta indica A. Juss). Plants were grown for 10 months in pots filled with soil having different salinity levels of 3, 6, 9, 12, 15, and 18 dS m−1 and compared with a control of 1.7 dS m−1. The results showed that plant growth and chlorophyll contents declined as salinity levels increased. Due to oxidative stress, the contents of H2O2 increased under higher salt levels. The mitigation of oxidative stress was achieved through the activation of antioxidant enzymes (catalase, ascorbate peroxidase, peroxidase, and superoxide dismutase). Multivariate analysis indicated that Na+ accumulation in plants was positively related to H2O2 production and enzymatic activities, and negatively related to plant biomass, chlorophyll contents, root and shoot K+ concentration, and root K+/Na+. The experimental results suggest that neem plants can be grown in moderate saline soils.

P-094 Genetic variations of antioxidant genes may coordinate with lifestyle to define male fertility: A cross-sectional study with control group

Abstract Study question Are polymorphisms of the oxidative stress-related genes different between infertile men and reproductively healthy men? Does lifestyle contribute to fertility outcomes? Summary answer A strong association between the polymorphisms of two antioxidant genes and sperm quality was identified. The infertile men had higher BMI and alcohol usage. What is known already Antioxidant genes including catalase (CAT), superoxide dismutase (SOD), and nitric oxide synthase (NOS) play a critical role in spermatogenesis and sperm functions, especially in stress-response cell signaling pathways. Polymorphisms of antioxidant genes have been reported to be associated with gene function, hence sperm quality, ultimately affecting male fertility in various cohorts. Body mass index (BMI) is universally considered a marker of health reflecting whole health, nutrition, and lifestyle. Smoking and alcohol consumption expose the body to harmful chemicals and oxidative stress. Here, we examined the polymorphisms of antioxidant genes in Vietnamese men and the link with lifestyle to regulate fertility. Study design, size, duration This cross-sectional study with control group recruited from Hanoi Medical University Hospital from 2018-2020. The participants were divided into two groups regarding semen analysis based on WHO 2010 criteria. The participants gave consent, then were clinically examined, interviewed for lifestyle, and genetically analyzed for antioxidant gene variations. An extra inclusion criterion for the control men was having at least one healthy baby. The exclusion criteria were chromosome aberrations and Y chromosome microdeletion. Participants/materials, setting, methods The control group (N = 85) and infertile group (N = 107) were recruited. Semen analysis was carried out by the CASA instrument based on WHO 2010 criteria. Lifestyle characteristics including exercise (per week), smoking (yes/no), alcohol consumption (the threshold for “yes” was five standard drinks/occasion, at least one occasion/month) were recorded. Four variants SOD1:7958G&amp;gt;A, SOD2:c.47T&amp;gt;C, CAT:-262C&amp;gt;T, and NOS3:-786C&amp;gt;T were analyzed by Sanger sequencing. Pearson’s, Chi-squared (χ2), Wilcoxon rank-sum, ANOVA test, and Bonferroni correction were applied where specifically required. Main results and the role of chance The average age of the two groups was not different (31.96 ± 4.87 vs 31.93 ± 6.3 years old, p = 0.92). The body mass index (BMI) of infertile men was significantly higher than the control (24.84 ± 2.31 vs 23.53 ± 2.55 kg/m2, p = 0.00026). No significant difference was identified regarding exercise and smoking. Notably, 96.26% of infertile men consumed alcohol in comparison with 88.23% of the control (p = 0,034). Genetically, SOD1:7958GA genotype and NOS3:-786CT genotype in the infertility group were significantly higher than in the control, OR = 2.191 (95% CI: 1.226–3.915, p = 0.004) and OR = 3.135 (95% CI: 1.591–6.180, p &amp;lt; 0.001), respectively. The frequency of the SOD2:c.47TC genotype was significantly higher in the male infertility group than in the control (OR = 1.941, 95% CI: 1.063–3.595, p = 0.029). Individuals carrying the SOD1:GA+AA, SOD2:TC+CC, and CAT:CT/TT genotype pattern had an increased risk of male infertility (OR = 7.614, p = 0.007), suggesting that gene-gene interaction plays an important role in male reproduction. To our knowledge, this is the first report worldwide on the association between the SOD1:7958G&amp;gt;A polymorphism and male infertility. Together, the data suggest that antioxidant gene polymorphisms and lifestyle, especially alcohol usage, coordinate to regulate male fertility. Limitations, reasons for caution Direct measurement of oxidative stress in the semen of men having different gene variations would be required to ascertain the link between the polymorphisms of antioxidant genes and fertility. The sample size was relatively small, a larger sample size would be required to accurately represent the Vietnamese population. Wider implications of the findings The findings can assist in infertility diagnosis focusing on the group of higher infertile risk genetic variations. Interventions such as antioxidant supplements would be developed and applied, lifestyle advice would be given specifically for a group having high-risk gene variations to accurately target fertility issues. Trial registration number Not applicable

Effects of Ultra-High-Pressure Treatment on Chemical Composition and Biological Activities of Free, Esterified and Bound Phenolics from Phyllanthus emblica L. Fruits

Phyllanthus emblica L. fruits (PEFs) were processed by ultra-pressure (UHP) treatment and then extracted by the ultrasonic-assisted extraction method. The influence of UHP on the phenolic composition, enzyme inhibitory activity and antioxidant activity of the free, esterified, and bound phenolic fractions from PEFs were compared. UHP pretreatment of PEFs significantly increased the total phenolic and flavonoid contents (p &lt; 0.05). A total of 24 chemical compositions were characterized in normal and UHP-treated PEFs by UHPLC-ESI-HRMS/MS. Compared with normal PEFs, these three different phenolic fractions had stronger antioxidant activities and inhibitory effects on the intracellular reactive oxygen species (ROS) production in H2O2-induced HepG2 cells (p &lt; 0.05). The ROS inhibition might be due to an up-regulation of the expressions of superoxide dismutase (SOD) and glutathione (GSH) activities. In addition, these three different phenolic fractions also significantly inhibited the activities of metabolic enzymes, including α-glucosidase, α-amylase and pancreatic lipase. This work may provide some insights into the potential economics and applications of PEFs in food and nutraceutical industries.

A Se Nanoparticle/MgFe-LDH Composite Nanosheet as a Multifunctional Platform for Osteosarcoma Eradication, Antibacterial and Bone Reconstruction.

Despite advances in treating osteosarcoma, postoperative tumor recurrence, periprosthetic infection, and critical bone defects remain critical concerns. Herein, the growth of selenium nanoparticles (SeNPs) onto MgFe-LDH nanosheets (LDH) is reported to develop a multifunctional nanocomposite (LDH/Se) and further modification of the nanocomposite on a bioactive glass scaffold (BGS) to obtain a versatile platform (BGS@LDH/Se) for comprehensive postoperative osteosarcoma management. The uniform dispersion of negatively charged SeNPs on the LDH surface restrains toxicity-inducing aggregation and inactivation, thus enhancing superoxide dismutase (SOD) activation and superoxide anion radical (·O2 -)-H2O2 conversion. Meanwhile, Fe3+ within the LDH nanosheets can be reduced to Fe2+ by depleting glutathione (GSH) in the tumor microenvironments (TME), which can catalyze H2O2 into highly toxic reactive oxygen species. More importantly, incorporating SeNPs significantly promotes the anti-bacterial and osteogenic properties of BGS@LDH/Se. Thus, the developed BGS@LDH/Se platform can simultaneously inhibit tumor recurrence and periprosthetic infection as well as promote bone regeneration, thus holding great potential for postoperative "one-stop-shop" management of patients who need osteosarcoma resection and scaffold implantation.

Effect of silicon spraying on rice photosynthesis and antioxidant defense system on cadmium accumulation

Cadmium (Cd) pollution is a serious threat to food safety and human health. Minimizing Cd uptake and enhancing Cd tolerance in plants are vital to improve crop yield and reduce hazardous effects to humans. In this study, we designed three Cd concentration stress treatments (Cd1: 0.20 mg·kg−1, Cd2: 0.60 mg·kg−1, and Cd3: 1.60 mg·kg−1) and two foliar silicon (Si) treatments (CK: no spraying of any material, and Si: foliar Si spraying) to conduct pot experiments on soil Cd stress. The results showed that spraying Si on the leaves reduced the Cd content in brown rice by 4.79–42.14%. Si application increased net photosynthetic rate (Pn) by 1.77–4.08%, stomatal conductance (Gs) by 5.27–23.43%, transpiration rate (Tr) by 2.99–20.50% and intercellular carbon dioxide (CO2) concentration (Ci) by 6.55–8.84%. Foliar spraying of Si significantly increased the activities of superoxide dismutase (SOD) and peroxidase (POD) in rice leaves by 9.84–14.09% and 4.69–53.09%, respectively, and reduced the content of malondialdehyde (MDA) by 7.83–48.72%. In summary, foliar Si spraying protects the photosynthesis and antioxidant system of rice canopy leaves, and is an effective method to reduce the Cd content in brown rice.

The Effects of ThymusCaramanicus Jalas Extract and Its Main Constituent Carvacrol Against Cisplatin-Induced Nephrotoxicity in Mice

Background: Cisplatin, an anti-cancer chemotherapy drug, has nephrotoxic effects. Thymus caramanicus Jalas (TCJ) has antioxidant effects due to its main components. Objectives: In the current research, we assessed the impact of TCJ extract and its main compound on cisplatin-induced nephrotoxicity in mice. Methods: Forty-two male mice were used in the study. Depending on their group, the animals received saline, carvacrol (10 mg/kg), or TCJ extract (50, 100, and 150 mg/kg) for 10 days. On the fifth day, mice received cisplatin (7.5 mg/kg, i.p.). After 10 days, serum creatinine (Cr) and blood urea nitrogen (BUN) levels were measured. Additionally, malondialdehyde (MDA) and glutathione (GSH) contents, as well as the activity levels of superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx), and total antioxidant capacity (TAC) were measured in the kidney tissues. The western blotting method was used to determine the kidney's expression of cleaved caspase-3, Bax, Bcl-2, nuclear factor kappa-B (NF-κB), and tumor necrosis factor-alpha (TNF-α). Kidney tissue damage score (KTDS) was assessed using the hematoxylin-eosin (H&amp;E) staining method. Results: Cisplatin significantly increased serum Cr, KTDS, MDA, BUN levels, NF-κB, TNF-α, cleaved caspase-3, and Bax protein expression in the cisplatin group compared to the control group (P &lt; 0.01). Additionally, cisplatin significantly decreased the kidney tissue's TAC and GSH content, activity levels of SOD, catalase, and GPx indicators, and expression of Bcl-2 protein (P &lt; 0.05). TCJ and carvacrol significantly ameliorated these indicators in the cisplatin + TCJ (150 mg/kg) and cisplatin + carvacrol (10 mg/kg) groups compared to the cisplatin group (P &lt; 0.05). Conclusions: TCJ (150 mg/kg) and its main component, carvacrol, could somewhat reduce cisplatin-induced nephrotoxicity through their anti-inflammatory, antioxidant, and anti-apoptotic effects.

P-368 In vitro culture with rutin and VEGF-A before xenotransplantation protects ovarian tissue from ischemia and oxidative damage but not from growing follicle depletion

Abstract Study question Can pre-treatment with antioxidant and vascular endothelial factor (VEGF-A) help to maintain healthy tissue structure and follicle pool in xenotransplanted human ovarian tissue? Summary answer Rutin and VEGF-A help to prevent fibrosis and oxidative stress damage in grafted ovarian tissue but cannot preserve growing follicles. What is known already Nowadays, ovarian tissue (OT) cryopreservation and autotransplantation become established fertility preservation methods in numerous countries. However, even if hundreds of children has been born thanks to this procedure, most crucial process during which 70-80% of the follicles are lost in the graft is the transplantation itself. As the graft restores vascularisation and regains its function the tissue undergoes ischemia and oxidative stress. There have been attempts made to improve this critical postoperative period by administration of different growth factors, hormones, antoxidants or mechanical tissue damage. Study design, size, duration Pieces of frozen-tharwed OT (2,5 × 2,5 mm) from 10 women were transplanted into immunodeficient mice for 4 weeks. Before xenotransplantation OT fragments were in vitro cultured for 36h in culture media supplemented with 1 μg/ml rutin and 50 ng/ml VEGF-A (n = 10 OT, pretreated grafted) or without (n = 10 OT, non-pretreated grafted). Frozen-thawed OTs were used as non-grafted controls for ischemia-reperfusion injury, apoptosis and follicle loss evaluation. Participants/materials, setting, methods OTs were received from cancer patients (n = 10) undergoing cryopreservation at National Cancer Institute. Xenotransplantation was performed at Life Sciences Centre, Vilnius University. Retrieved grafts were devided into one small piece for immunochistochemical evaluation of α-SMA and apoptosis by TUNEL, and one bigger piece for qPCR analysis of genes related to tissue integrity (PRDX1, SOD1, FN1, COL6A1, CASP3, ANGPT2, mouse CD31) and follicle development (FOXO3, FSHR, GDF9), house keeping gene being GAPDH/Gapdh. Main results and the role of chance After 4 weeks of grafting, multiple genes and protein expressions were evaluated. Vascularisation evaluation: compared to non-grafted group, in grafted OTs we observed significantly more vessels (α-SMA staining, p &amp;lt; 0.0001 and ANGPT2 expression, p &amp;lt; 0.05), as well 1,5-fold higher relative expression of mouse CD31 in pretreated grafted group vs non-pretreated grafted group. Qxidative stress and apoptosis evaluation: expression of SOD2, PRDX1 and CASP3 genes was lower in both grafted groups compared to non-grafted control, significant alterations between xenotransplanted groups were found. Structural gene evaluation: in non-grafted control we could find higher expression of COL6A1 and FN1 genes compared to grafted groups and statistical difference of COL6A1 expression in between grafted groups (p &amp;lt; 0.05). Follicle pool evaluation: primordial follicle activation was measured by FOXO3 gene expression change, though we did not find significant differences between any of three groups, p = 0.06 was observed between grafted OT groups. Maturing follicles were evaluated by expression changes in GDF9 and FSHR genes. We observed significant decrese in expression of these genes in grafted groups (p &amp;lt; 0.05). Results from TUNEL are awaited, thus are the quantification of apoptotic cells, which will provide valuable insights for correlation with vascularisation and oxidative stress. Limitations, reasons for caution This study was performed only in a long-term xenotransplantation setting, we cannot know if rutin and VEGF-A had a significant influence during the first days of OT transplantation. Due to the small size of OT grafted, counting and distribution of follicles was not performed, Wider implications of the findings This study shows that pretreatment of OTs with rutin and VEGF-A could be potentially used in human setting due to protection of ischemia and oxidative stress, especially if the survival of growing follicles is not required, as it is in cases of prepubertal girls. Trial registration number not applicable

P-082 Use of TetraSOD® in male infertility: double-blind randomized placebo-controlled study

Abstract Study question Can the use of TetraSOD® (microalgae with high content of the antioxidant enzyme superoxide dismutase) improve semen quality and reduce oxidative stress in sperm? Summary answer The use of TetraSOD® reduces the oxidative stress of sperm, increasing the total sperm count in oligozoospermia and reducing the double-stranded DNA sperm fragmentation. What is known already Male factor is present in approximately 50% of the infertile couples, a large number of them of unknown or idiopathic origin. It has been demonstrated that semen quality is deteriorating over time, perhaps as a result of exposure to several environmental factors related with lifestyle that could impact both directly and indirectly on the complex process of spermatogenesis. Some of these bad habits of lifestyle increase the levels of oxidative stress in semen, which is associated with lower sperm concentration and motility. In this scenario, the use of antioxidants can be useful to increase sperm quality. Study design, size, duration Double-blind randomized placebo-controlled clinical study to assess the effect of 250 mg of TetraSOD® (microalgae with high content of the primary antioxidant enzyme superoxide dismutase). Eighty patients with idiopathic infertility showing asthenozoospermia, oligozoospermia or oligoasthenozoospermia were recruited and randomized to receive 250 mg of TetraSOD® (TSOD group) or placebo (Placebo group) for 3 months. Participants/materials, setting, methods Men with asthenozoospermia, oligozoospemia or aligoastenozoospermia and idiopatic infertility were recruited after informed consent signature. Patients were randomized into two groups. TetraSOD® group received 250 mg of TetraSOD®during 3 months, and Placebo group received capsules without TetraSOD® during the same period of time. Seminogram, double-strand sperm DNA fragmentation (COMET assay) and sORP (static oxidation- reduction potential) were studied at baseline (V0) and after 3 months (V3). Main results and the role of chance TetraSOD® consumption for three months elicited a statistically significant increase (Wilcoxon matched-pairs signed rank test; p=0.02) in the average number of sperms per ejaculate (from 17.06 x 106 to 41.94 x 106) in participants displaying oligozoospermia (&amp;lt;39 x 106 sperms) at V0. In contrast, the lower variation in the measured values in the Placebo group was not statistically significant. Regarding sORP, a statistically significant decrease (Wilcoxon matched-pairs signed rank test; p=0.0233) was observed in participants who displayed a value at V0 between 1.34-10. After TetraSOD® consumption, the average changed from 2.96 to 1.91 mv/106 sperm/ml from V0 to V3. However, not statistically significant changes were detected in the Placebo group between sORP values at V0 and V3. Moreover, in relation to double-stranded DNA breaks, a statistically significant decrease (Wilcoxon matched-pairs signed rank test; p=0.01) in the average percentages (from 31.81% to 25.93%) was measured after consumption of TetraSOD® at V3 in participants displaying a compromised redox status (sORP&amp;gt;1.34-10) at V0. No significant changes were found in the Placebo group. Limitations, reasons for caution The study was performed only in one ethnic group, so in future studies other ethnicities should be included. Wider implications of the findings TetraSOD® may improve male fertility because increases the number of sperms per ejaculate in oligozoospermia and reduces double-stranded DNA fragmentation in sperm when individuals display an altered redox status in semen. Such results might be related to the indirect antioxidant effect of TetraSOD®, reducing oxidative stress in seminal fluid. Trial registration number Not applicable

P-101 Mitochondria-mediated apoptosis induced testicular dysfunction in bisphenol F treated rats: ameliorative effect of baicalin

Abstract Study question Will baicalin treatment ameliorate bisphenol F (BPF)-induced testicular dysfunction? Summary answer Baicalin ameliorates BPF-induced mitochondrial dysfunction-associated apoptosis What is known already BPF is a chemical presently considered a major replacement for bisphenol A and has been widely used in the canning and other food processing industries. However, a major limitation of this chemical is its gonadotoxic effect. Studies have shown that BPF disrupts testicular function via apoptotic signaling, although, its mechanism of action has not been fully explored. On the other hand, baicalin is an anti-apoptotic agent. Yet, none of these studies has reported the effect of baicalin on BPF-induced testicular injury Study design, size, duration This is a prospective experimental study using an animal model. Twenty-four sexually matured male Wistar rats of comparable weight were used for this study. All treatments were via oral gavage and lasted for 8 weeks Participants/materials, setting, methods Animals were acclimatized for two weeks, then randomized into 4 groups (n = 6); control: animals received 0.5 mL of corn oil as vehicle, BPF: animals received 30 mg/kg of BPF, baicalin: animals treated with 100 mg/kg of baicalin, BPF+ baicalin: animals treated with 30 mg/kg BPF and 100 mg/kg baicalin Main results and the role of chance : Baicalin ameliorated BPF-induced decrease in spermatogenic indices and sperm quality, circulatory levels of follicle-stimulating hormone, luteinizing hormone, and testosterone, and testicular concentrations of 3β-HSD and 17β-HSD. Also, baicalin ameliorated BPF-induced decline in testicular superoxide dismutase, catalase, total thiol, glutathione, glutathione S-transferase, and glutathione peroxidase activities and nuclear factor erythroid 2-related factor 2 concentrations. Furthermore, baicalin alleviated BPF-induced increase in testicular malondialdehyde, myeloperoxidase, TNF-α, IL-6, and NF-kB levels. These events were associated with the dampening of BPF-induced decline in NAD+/NADH and Bcl-2 activities and BPF-induced increase in Bax and caspase 3 activities by baicalin. Limitations, reasons for caution This study was conducted in a rat model; therefore the results of the present study should be extrapolated to humans with caution Wider implications of the findings The present study described the ameliorative effect of baicalin treatment on BPF-induced testicular toxicity. Trial registration number N/A

In Vitro Protective Effect of Pea-Derived Peptides (PPs) via the Keap1/Nrf2 Signaling Pathway on Alpha-Gliadin-Sensitizing Peptide Induced Cacao-2 Cells.

Celiac disease (CD) is an allergic intestinal disease caused mainly by gliadin in wheat, which is widespread in the population and currently lacks effective treatment. α-Gliadin peptides cause cellular damage by substantially increasing cellular reactive oxygen species (ROS) levels. This study investigates the protective effect of 11 pea-derived peptides (PPs) on ɑ-gliadin peptide (P31-43) treated Caco-2 cells. Results show that cells treated with PP2, PP5, and PP6 peptides significantly reduce the cell mortality caused by P31-43. Three PPs significantly reduce the P31-43-induced decrease in ROS levels to control levels, and there is no difference between them and the vitamin C (Vc) group. The results in terms of antioxidant-related enzymes show that PPs significantly decrease superoxide dismutase activity (SOD), glutathione reductases (GR), and glutathione (GSH)/oxidized glutathione (GSSG) levels, thus significantly enhancing the antioxidant level of cells. By studying the key proteins of the Kelch-like ECH-associated protein 1 (Keap1)/NF-E2-related factor 2 (Nrf2) pathway, it is found that PPs activate the Keap1/Nrf2 signaling pathway. The study finds that peptides from peas can effectively alleviate ɑ-gliadin peptide-induced cell damage. The discovery of these food-derived peptides provides novel potential solutions for the prevention and treatment of CD.

Betaine alleviates doxorubicin-related cardiotoxicity via suppressing oxidative stress and inflammation via the NLRP3/SIRT1 pathway.

Cardiotoxicity is one of the side effects of the anti-cancer drug doxorubicin (DOX) that limits its clinical application. Betaine (BT) is a natural agent with promising useful effects against inflammation and oxidative stress (OS). We assessed the effects of BT on DOX-induced cardiotoxicity in mice. Forty-two male NMRI mice were assigned to six groups: I: control; II: BT (200mg/kg; orally, alone); III: DOX (2.5mg/kg; six injections (ip)) for two weeks; IV, V, VI: BT (50mg/kg, 100mg/kg, and 200mg/kg; orally, once a day for two weeks, respectively) plus DOX administration. The cardiac enzymes like cardiac troponin-I (cTn-I), lactate dehydrogenase (LDH), and creatine kinase-MB (CK-MB) were assessed in serum. Oxidative/inflammatory markers like nitric oxide (NO), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), reduced glutathione level (GSH), and glutathione peroxidase (GPx) activities were determined in cardiac tissue. The expressions of NOD-like receptor protein 3 (NLRP3), caspase-1, interleukin (IL)-1β, and silent information regulator 1 (SIRT1) proteins were also evaluated in cardiac tissue. The results indicated that DOX significantly increased LDH, CK-MB, cTn-I, MDA, and NO levels and also the caspase-1, NLRP3, and IL-1β expression. Furthermore, DOX caused a significant reduction in the GSH levels and SOD, CAT, GPX activities, and the expression of SIRT1 protein in heart tissue. However, BT significantly improved all studied parameters. The findings were confirmed by histopathological assessments of the heart. BT can protect against DOX-induced cardiotoxicity by suppressing the activation of NLRP3 and OS by stimulating the SIRT1 pathway.

Evaluation of the sperm parameters, oxidative stress, and histopathological effects of vitamin B12 in preventing Helicobacter pylori-induced testicular toxicity: An experimental study

Background: Helicobacter pylori (H. pylori) causes sterility by affecting the reproductive system. vitamin B12 improves sperm quantity and function. Objective: Vitamin B12 protection against H. pylori adverse effects was investigated. Materials and Methods: 40 C57 male mice (6 wk) were randomly assigned to 4 equal groups (n = 10) including, group 1 (control without any intervention), group 2 (H), 3 (HP), and 4 (HB) received 1×109 colony forming unit (CFU) of H. pylori, 1×109 CFU of H. pylori+phosphate buffered saline, 1×109 CFU of H. pylori+50 μg/kg vitamin B12 intraperitoneally, respectively. In the induction groups, the H. pylori was orogasterically injected 3 times with 1 cc phosphate buffered saline throughout the day. Then testicular metrics, sperm motility, viability, quantity, and shape, plasma levels of malondialdehyde (MDA), superoxide dismutase, glutathione peroxidase, and total antioxidant capacity were measured. Also, testicular-tissue changes were examined using Johnson scores, tubular differentiation index, and spermatogenesis index. Vitamin B12, homocysteine, and testosterone serum levels were examined. Results: The results showed a significantly lower Johnson score, tubular differentiation index, and spermatogenesis index, and serum level of testosterone and homocysteine as well as a higher MDA level in the H and HP groups than the HB group (p &lt; 0.05). In contrast, the highest superoxide dismutase and glutathione peroxidase enzymes activity and total antioxidant capacity as well as the lowest serum level of MDA were found in the HB group compared to other groups (p &lt; 0.05). Conclusion: Vitamin B12 increased antioxidant enzyme activity, enhanced sperm parameters, and decreased injury to testicular tissue. It can be used as a potent antioxidant in reducing testicular damage induced by H. pylori. Key words: Helicobacter pylori, Reproductive medicine, Oxidative stress.

Dietary effects of vitamin C on antioxidant capacity, intestinal microbiota and the resistance of pathogenic bacteria in cultured Silver pomfret (Pampus argenteus).

As most teleosts are unable to synthesize vitamin C, supplemental diets containing vitamin C diets play a crucial role in fish health. The aim of this study was to investigate the effect of dietary vitamin C on the intestinal enzyme activity and intestinal microbiota of silver pomfre (Pampus argenteus). Four experimental diets were supplemented with basic diets containing 300 mg of vitamin C/kg (group tjl3), 600 mg of vitamin C/kg (group tjl6), and 1200 mg of vitamin C/kg (group tjl12), as well as vitamin C-free supplemental basic diet (group tjl0), respectively. The four diets were fed to juvenile P. argenteus (average initial weight: 4.68 ± 0.93 g) for 6 weeks. The results showed that the activity of SOD (superoxide dismutase) and CAT (catalase) increased significantly while that of MDA (malondialdehyde) decreased significantly in group tjl3 compared to vitamin group tjl0. At the genus level, groups tjl0, tjl6, and tjl12 contained the same dominant microbial community, Stenotrophomonas, Photobacterium, and Vibrio, whereas group tjl3 was dominated by Stenotrophomonas, Delftia, and Bacteroides. Among the fish fed with a basic diet containing 300 mg of vitamin C/kg, the intestines exhibited a notable abundance of probiotic bacteria, including lactic acid bacteria (Lactobacillus) and Bacillus. The abundance of Aeromonas in groups tjl3 and tjl6 was lower than that of the vitamin C-free supplemental basic diet group, whereas Aeromonas was not detected in group tjl12. In addition, a causative agent of the disease outbreak in cultured P. argenteus, Photobacterium damselae subsp. Damselae (PDD) was the dominant microbiota community in groups tjl0, tjl6 and tjl12, whereas the abundance of PDD in group tjl3 was the lowest among the diets. Taken together, the diets supplied with vitamin C could influence the composition microbial community of P. argenteus. The low level of vitamin C (300 mg of vitamin C/kg per basic diet) supplementation could not only improve the antioxidant capacity but also resist the invasion of pathogenic bacteria.