Amount Of GSH Research Articles

The Effects of Glutathione Monoethyl Ester on Different Biochemical, Oxidant, and Antioxidant Levels During Storage in Leukoreduced Red Blood Cells

Background: It is essential to maintain the quality of the stored blood, because various factors affect the stored red blood cells (RBCs) over time, some red blood cell storage lesions (RCSL) develop during storage, and it could reduce the function of the RBCs. The present study aimed to evaluate the effects of glutathione monoethyl ester on different biochemical changes, oxidant, and antioxidant levels in the leukoreduced RBCs (LR-RBCs) during storage. Materials and Methods: About 10 units of LR-RBC were collected, processed and stored according to the standard operating procedures (SOPs) of the Iranian Blood Transfusion Organization. Each unit divided into 2 equal parts; LR-RBC treated with glutathione monoethyl ester and a control group. Exposure of phosphatidylserine (PS), reactive oxygen species (ROS) and microvesicle derived from the RBCs (RBC-MVs), were measured by the flow cytometry method. ELISA was used to measure the level of glutathione, and 2, 3-diphosphoglycerate (2,3-DPG). Glucose-6-phosphate dehydrogenase (G6PD) enzyme activity was measured with a chemistry autoanalyzer. Results: The levels of glutathione reduced the initial value in the treated group (80%), and the control group (60%), respectively. Exposure of surface PS, ROS and RBC-MVs increased significantly during storage time for consecutive weeks to the amount of GSH. The levels of 2,3-DPG decreased with increasing storage time. Conclusions: Overall, The study suggest that glutathione monoethyl ester is effective to reduce the oxidative stress and the quality of RBCs can be improved.

Ethylene and hydrogen sulfide regulate hexavalent chromium toxicity in two pulse crops: Implication on growth, photosynthetic activity, oxidative stress and ascorbate glutathione cycle

Sustainable agriculture has become prime importance to feed growing population. To achieve this goal application of exogenous hormones and signaling molecules are gaining important. In this context, we have investigated potential of ethylene (25 μM ethephon; donor) and H2S (10 μM NaHS; donor) in mitigating hexavalent chromium [Cr (VI), 50 μM] toxicity in two pulse seedlings: black bean and mung bean. Cr(VI) declined growth and gas exchange parameters (photosynthetic rate, stomatal conductance, sub cellular CO2 concentration, and transpiration level) which was accompanied by intracellular accumulation of Cr in both pulse crops and the damaging effect was greater in mung bean seedlings. The suppression in the growth and related parameters was occurred due to higher buildup of oxidative stress markers; O2•‾, H2O2, lipid peroxidation (as malondialdehyde, MDA equivalents) and membrane injury in leaf and root of both pulse crops. Cr induced disturbance in AsA-GSH cycle (reduction in the activity of glutathione reductase, ascorbate peroxidase, monodehydroascorbate reductase and dehydroascorbate reductase, and the amount of ASA and GSH) could be one of the reasons for greater accumulation of H2O2. Further, exogenous application of ethylene and H2S significantly ameliorated Cr toxicity on growth and photosynthetic activity by significantly lowering the intracellular Cr accumulation and oxidative biomarkers, and also by strengthening the activity of AsA-GSH cycle. The exogenous application of biosynthesis inhibitors of ethylene (AVG) and H2S (PAG) caused greater damaging effect on these parameters due to more accumulation of Cr(VI), thereby suggesting that the endogenous levels of these regulators are critical for Cr(VI) tolerance. Interestingly, ET did not rescue adverse effects of Cr(VI) in absence of endogenous H2S, while H2S could do so even without endogenous ethylene, suggesting that H2S played downstream signaling to ethylene in regulating Cr(VI) toxicity. Hence, being cheap and easily available theses growth regulators may be considered for sustainable agriculture.

Effect of Preservation Methods on Fat-Soluble Vitamins and Stress Biomarkers in Rhus coriaria L. (Sumac) of Different Regions

The number of fat-soluble vitamins and stress biomarkers in Rhus coriaria L. samples taken from different regions was determined by HPLC before and after being subjected to different preservation methods. For this purpose, one group of the samples was analyzed immediately, while the other two groups one of which oiled, and the other group is kept as is for six months. It was determined that the amounts of vitamin A, E, β-carotene and lycopene in fresh sumac samples varied between 1.12 - 2.77, 84.40 - 230.65, 2.48 - 5.31 and 8.10 - 26.90 µg (g dw)-1, respectively. The highest loss of vitamins was observed in an unoiled group of samples. The amounts of GSH, GSSG, MDA, 4-HNE, and GSH/GSSG in the same samples varied between 1004.12 - 2550.42, 422.54 - 1375.38, 13.95 - 31.30, 7.12 - 15.40 µg (g dw)-1, and 1.16 - 3.49, respectively. While the amount of GSH and GSH/GSSG ratio in the stored sumac samples for six months decreased, on the other hand amount of MDA, GSSG, and 4-HNE increased. Differences in all examined parameters in fresh, unoiled, and oiled sumac samples are statistically significant (P

Development of a Golgi apparatus-targeting probe for revealing the fluctuations of GSH in the living cells during ferroptosis

Ferroptosis is a distinctive iron-dependent and GSH-related cell death form, and has been demonstrated to closely relate with Golgi apparatus (GA). Exploring the fluctuations of GSH level in GA during ferroptosis is of great importance for the deep research of biological functions of GA in ferroptosis. Herein, we reported a novel GA-targeting probe (NA-G) for the detection of GSH in GA during ferroptosis. The probe NA-G utilized N-oxidized pyridine sulfonyl as a responsive site for GSH, with tetradecanoyl amide serving as GA-targeting moiety. NA-G exhibited remarkable sensitivity and selectivity, manifesting a heightened response to GSH and effectively visualizing GSH through bright green fluorescence under UV light. By fluorescence imaging of cellular GSH, NA-G was demonstrated to serve as a GA-targeting probe, and could successfully monitor the dynamic changes in intracellular GSH. Further experimentation revealed that NEM stimulation resulted in the removal of a significant amount of endogenous GSH in GA, and erastin-induced ferroptosis significantly reduced GSH levels in GA. Notably, we discovered that Fer-1 and VE effectively inhibited the substantial depletion of GSH in GA during ferroptosis.

Hepatoprotective effect of amifostine and WR-1065 on acetaminophen-induced liver toxicity on Wistar rats.

The most important problem with acetaminophen is its hepatotoxicity. N-acetylcysteine (NAC) is used to treat the hepatotoxicity of acetaminophen. Due to the structural similarities of this compound with amifostine, we decided to test the effect of this substance and its metabolite, WR-1065, on the hepatotoxicity of acetaminophen. The single-dose method contained 1. Control; 2. Acetaminophen (1 g/kg, gavage); 3-5. Acetaminophen + amifostine (100, 200, 400 mg/kg, i.p.); 6-8. Acetaminophen + WR-1065 (50, 100, 200 mg/kg, i.p.); and 9. Acetaminophen + NAC (100, 200 mg/kg, i.p.). The multiple-dose method included the same groups: amifostine (50, 100, 200 mg/kg), WR-1065 (25, 50, 100 mg/kg), and NAC (100 mg/kg). Then, animals were sacrificed, and blood samples were collected for measuring ALT, AST, ALP, and T-Bil, liver tissue for histopathological examination, MDA, and GSH amounts. Acetaminophen increased the levels of MDA, T-Bil, ALT, AST, and ALP, decreased GSH levels, and augmented necrosis, neutrophils, lymphocytes, and macrophages in the port space in single-dose and multiple-dose studies. Amifostine and WR-1065 significantly reduced the levels of MDA, T-Bil, ALT, AST, ALP, increased GSH content, and ameliorated histopathological alterations in a single-dose and multiple-dose method compared to the acetaminophen group. Moreover, NAC caused a significant decrease in the levels of MDA, T-Bil, ALT, AST, and ALP, and reduced GSH amounts in single-dose and multiple-dose studies. Amifostine and WR-1065 as antioxidant and hepatoprotective compounds are effective in reducing acetaminophen-induced hepatotoxicity with a similar effect to NAC and can be administered as an adjunct in the treatment of acetaminophen overdose.

Hydrolytic degradation and in vitro studies of poly(alkylene citrate)-based polyesters modified with glutathione

Materials currently used for vascular grafts are characterized by limited biocompatibility. Especially challenging is obtaining the degradable graft to regenerate a patient's native tissue. Poly(alkylene citrates) (PACs) are promising for soft tissue engineering applications; however, their degradation must follow a specific profile and must not result in the formation of any substances cytotoxic to the human body. The aim of this study was to evaluate the processes that occur throughout the hydrolytic degradation of PACs with a 2:3 M ratio of citric acid and diol, additionally modified with an antioxidant compound: glutathione (GSH). The samples were tested after various time intervals of incubation in an aqueous environment (up to 3 months), taking into account the influence of the type of diol used (1,6-hexanediol or 1,8-octanediol), the time of polymer cross-linking (4 or 10 days) and the presence of a different amount of GSH (0.4 or 0.8 wt%). Structural changes in the samples were monitored by means of swelling capacity, mass changes, hardness tests, mass spectrometry, and UV spectrofluorimetry. After 3 months of degradation, a higher weight loss was observed for the samples based on 1,6-hexanediol (>20 wt%) than those based on 1,8-octanediol (6–16 wt%). Based on the values obtained, the approximate time of 80% of the mass loss was calculated, which was higher for samples cross-linked for 10 days than 4 days. The samples based on 1,8-octanediol degraded slower than those of 1,6-hexanediol, but their degradation pattern was more linear and the amount of heavy ions present in the supernatant was negligible. In vitro studies on L929 fibroblasts did not show a cytotoxic effect of the materials tested. The study proved that GSH does not cause significant changes in the degradation profiles, sufficiently providing the fluorescence properties of the materials. Furthermore, any drastic changes in the material for up to 3 weeks followed by progressive degradation may support the tissue before the regeneration process begins.

Effect of Plant Growth-Promoting Bacteria on Antioxidant Status, Acetolactate Synthase Activity, and Growth of Common Wheat and Canola Exposed to Metsulfuron-Methyl.

Metsulfuron-methyl, a widely used herbicide, could cause damage to the sensitive plants in crop-rotation systems at extremely low levels in the soil. The potential of plant growth-promoting bacteria (PGPB) for enhancing the resistance of plants against herbicide stress has been discovered recently. Therefore, it is poorly understood how physiological processes occur in plants, while PGPB reduce the phytotoxicity of herbicides for agricultural crops. In greenhouse studies, the effect of strains Pseudomonas protegens DA1.2 and Pseudomonas chlororaphis 4CH on oxidative damage, acetolactate synthase (ALS), enzymatic and non-enzymatic antioxidants in canola (Brassica napus L.), and wheat (Triticum aestivum L.) were investigated under two levels (0.05 and 0.25 mg∙kg-1) of metsulfuron-methyl using spectrophotometric assays. The inoculation of herbicide-exposed wheat with bacteria significantly increased the shoots fresh weight (24-28%), amount of glutathione GSH (60-73%), and flavonoids (5-14%), as well as activity of ascorbate peroxidase (129-140%), superoxide dismutase SOD (35-49%), and ALS (50-57%). Bacterial treatment stimulated the activity of SOD (37-94%), ALS (65-73%), glutathione reductase (19-20%), and the accumulation of GSH (61-261%), flavonoids (17-22%), and shoots weight (27-33%) in herbicide-exposed canola. Simultaneous inoculation prevented lipid peroxidation induced by metsulfuron-methyl in sensitive plants. Based on the findings, it is possible that the protective role of bacterial strains against metsulfuron-metil is linked to antioxidant system activation.

Renoprotective effect of thymoquinone against rhabdomyolysis-induced acute kidney injury in the rat model.

Rhabdomyolysis leads to the release of myoglobin, sarcoplasmic proteins, and electrolytes into the blood circulation causing acute kidney injury (AKI). Thymoquinone, a natural compound found in Nigella sativa seeds, has antioxidant and anti-inflammatory effects. This investigation assessed the renoprotective effect of thymoquinone on rhabdomyolysis-induced AKI in rats. Male Wistar rats were categorized into six groups (n = 6): 1. Control: (normal saline), 2. Glycerol (50 ml/kg, single dose, IM), 3-5: Glycerol + thymoquinone (1, 2.5 and 5 mg/kg, 4 days, IP), 6. Thymoquinone (5 mg/kg). On day 5, serum and kidney tissue were isolated and the amounts of serum creatinine and blood urea nitrogen (BUN), renal malondialdehyde (MDA), glutathione (GSH.), tumor necrosis factor-alpha (TNF-α), neutrophil gelatinase-associated lipocalin (NGAL), and pathological changes were evaluated. Glycerol increased creatinine, BUN, MDA, TNF-α, and NGAL levels. It decreased GSH amounts and caused renal tubular necrosis, glomerular atrophy, and myoglobin cast in kidney tissue. Co-administration of glycerol and thymoquinone reduced creatinine, BUN, histopathological alterations, and MDA levels, and enhanced GSH amounts. Administration of glycerol and thymoquinone (5 mg/kg) had no significant effect on TNF-α amount but decreased NGAL protein levels. The administration of thymoquinone (5 mg/kg) alone did not display a significant difference from the control group. Rhabdomyolysis from glycerol injection in rats can cause kidney damage. Thymoquinone may attenuate renal dysfunction and oxidative stress. However, the TNF-α level was not significantly affected. Further studies are needed to explore the potential therapeutic effects of thymoquinone in managing AKI.

Synergistic gastroprotective and antioxidative effects of natural olive oil and usnic acid isolated from Usnea longissima, a lichen species in Anatolia (Türkiye), in the indomethacin ulcer model created in rats

Usnea longissima, a medically important lichen growing up in forests in Anatolia (Turkey). In this study, the gastroprotective effect of usnic acid (UA) was investigated using an indomethacin (IND)-induced gastric ulcer model in rats. While 25, 50, 100 and 200 mg/kg UA doses were dissolved in 2 ml of olive oil (OO) and administered to rats, only OO was given to one group. In addition, lansoprazole (LAN) and ranitidine (RAN) and IND were dissolved in water and administered to rat groups. IND administration caused very high levels of damage to rat stomachs. On the other hand, when four doses of UA, OO, RAN and LAN were administered, it was determined that hyperemias in the stomach of rats was significantly reduced. After macroscopic analysis of gastric tissues, the activities of superoxide dismutase (SOD), catalase (CAT), myeloperoxidase (MPO) and nitric oxide synthase (iNOS and cNOS) enzymes as well as glutathione (GSH) and lipid peroxidation (LPO) levels were determined in these tissues. After IND application, it was detected increases in MPO, CAT and iNOS activities in gastric tissues and decreases in SOD, cNOS and GSH amounts. Four doses of UA, OO, RAN and LAN applications reversed the trend, bringing them closer to healthy levels.

Clusterin is upregulated by erastin, a ferroptosis inducer and exerts cytoprotective effects in pancreatic adenocarcinoma cells.

Ferroptosis is a novel form of cell death, which is distinguished from apoptosis and necrosis, and characterized by accumulation of lipid-based reactive oxygen species (ROS) in an iron-dependent manner. Erastin, a small molecule, was widely reported to trigger ferroptosis in various kinds of cancer cells, including pancreatic cancer cells by inducing ROS accumulation. However, how erastin treatment exerts cytotoxicity is not still fully understood. In this study, the effects of erastin in causing pancreatic cancer cell death via inducing ferroptosis and apoptosis are investigated. As expected, erastin treatment caused ROS accumulation, increase in iron concentration and non-apoptotic cell death, which is different from that of induced by apoptosis inducer, staurosporine. Interestingly, erastin treatment caused the upregulation of clusterin, which contributes to the regulation of malignant behaviors of pancreatic cancer, including preventing apoptosis and inducing chemoresistance. Without erastin treatment, overexpressed clusterin significantly promoted cell proliferation, which is consistent with its cytoprotective roles. After erastin treatment, overexpressed clusterin decreased erastin-induced ROS accumulation and cell death. By measuring iron concentration, reduced glutathione (GSH) and glutathione peroxidase 4 (GPX4), it is revealed that clusterin caused resistance to erastin-induced ferroptosis potentially via maintaining the enzymatic activity of GPX4, without disturbing GSH amount. Thus, ferroptosis inducer, erastin, may crosstalk with apoptotic cell death via regulating clusterin, indicating a more complex regulatory network between ferroptosis and apoptosis.

An off–on fluorescence sensor based on hydrogen-bonded organic frameworks for the detection of glutathione

Hydrogen-bonded organic frameworks (HOFs) have drawn great attention in recent years; however, few reports show the applications of HOFs in biosensing. Abnormal concentration of glutathione (GSH) usually relates to some diseases, thus quantifying the amount of GSH is of great importance. Fluorescence method has the advantages of high sensitivity, simplicity, and low cost, however, traditional fluorescent sensors for GSH need complex synthesis procedures for fluorescent probes. Herein, a simple off–on fluorescence sensor based on hydrogen-bonded organic frameworks (HOF-PyTTA) was constructed. Firstly, the fluorescence of HOF-PyTTA was quenched by Fe3+, and after the addition of GSH, Fe3+ reacted with GSH to restore the fluorescence intensity of HOF-PyTTA. Based on the above principles, a simple HOF-PyTTA/Fe3+ mixed system was constructed and applied to detect GSH. The concentration of GSH showed a good linear relationship with the fluorescent intensity of HOF-PyTTA/Fe3+ in the range of 0–0.40 mmol/L, and the limit of detection was 8.8 μmol/L. This method was applied to the determination of GSH in human serum samples. The recoveries were in the range of 95.9–102.5 %, and the RSD was 0.1 % − 0.64 %. The proposed method based on the off–on fluorescence change of HOF-PyTTA shows a simple operation and a rapid response within 10 min. It not only broadened the application of hydrogen-bonded organic frameworks but also indicated a great potential of HOF as an efficient fluorescence probe for biosensing.

Efficacy and Potential Mechanism of Essential Oils of Three Labiatae Plants against the Pathogenic Fungi of Root Rot Disease in Atractylodes chinensis

Atractylodes chinensis has a high medicinal value and is widely cultivated. However, root rot disease seriously affects the yield and quality of A. chinensis. To develop green and safe pesticides, the inhibitory effect of essential oils (EOs) of three Labiatae plants on the pathogenic fungi that causes root rot disease in Atractylodes chinensis was investigated. The results showed that the Origanum vulgare EO and Thymus mongolicus EO exhibited strong inhibitory effects on Fusarium oxysporum, Fusarium solani, and Fusarium redolens, with 100% inhibition rate. The low MIC values of EOs and their main components against the three pathogenic fungi indicated that all of them showed strong fungicidal effects. The MIC values of O. vulgare EO against F. oxysporum, F. solani, and F. redolens were 2.60 mg/mL, 3.13 mg/mL, and 1.56 mg/mL, respectively. Analyses using scanning electron microscopy (SEM) and transmission electron microscopy (TEM) showed that the O. vulgare EO severely damaged the cell wall and cell membrane of mycelial cells. The O. vulgare EO increased cell permeability, leading to a large leakage of cell contents (DNA and proteins). In addition, O. vulgare EO inhibited F. oxysporum by inducing ROS production and reducing the amount of intracellular GSH, leading to a large accumulation of ROS. This study showed that plant EOs have excellent fungicidal activity and can be used as novel natural and environmentally friendly pesticides for the control of root rot in A. chinensis.

Vitamin D ameliorates Aeromonas hydrophila-induced iron-dependent oxidative damage of grass carp splenic macrophages by manipulating Nrf2-mediated antioxidant pathway

Aeromonas hydrophila (A. hydrophila) is one of major pathogenic bacteria in aquaculture and potentially virulent to grass carp (Ctenopharyngodon idella). As an essential nutrient for fish, vitamin D3 (VD3) has been reported to play a role against oxidative stress, but the exact mechanism remains to be elusive. In this study, we found that A. hydrophila induced ferrugination and macrophage aggregation in the spleen of grass carp. Along this line, using the splenic macrophages as the model, the effects of VD3 on A. hydrophila-caused iron deposition and subsequent injuries were determined. In the context, 1,25D3 (the active form of VD3) significantly reduced cellular free Fe2+, lipid peroxidation and lactic dehydrogenase (LDH) release induced by A. hydrophila in the splenic macrophages, indicating the protective effects of VD3 on A. hydrophila-led to ferroptosis-related injuries. In support of this notion, 1,25D3 was effective in hindering ferroptosis inducers-stimulated LDH release in the same cells. Mechanically, 1,25D3 enhanced iron export protein (ferroportin1) and glutathione peroxidase 4 (GPX4) protein levels, and glutathione (GSH) contents via vitamin D receptor (VDR). Moreover, NF-E2-related factor 2 (Nrf2) pathway mediated the regulation of 1,25D3 on GPX4 protein expression and GSH synthesis. Meanwhile, 1,25D3 maintained the stability of Nrf2 proteins possibly by attenuating its ubiquitination degradation. Furthermore, in vivo experiments showed that 1,25D3 injection could not only improve the survival of fish infected by A. hydrophila, but also enhance GSH amounts and decrease malonaldehyde (MDA) contents and iron deposition in the spleen. In summary, our data for the first time suggest that VD3 is a potential antioxidant in fish to fight against A. hydrophila induced-ferroptotic damages.

Generation of kokumi γ-glutamyl short peptides in Spanish dry-cured ham during its processing

The typical dry-cured ham flavor is rich in umami and brothy perceptions, for which short peptides may contribute. Particularly, γ-glutamyl peptides could be the responsible of these previously reported attributes, as they exert a synergistic interaction with other basic tastes and modify the intensity of salty, sweet, and umami tastes. The content of peptides has been reported to evolve along the processing, but no kokumi γ-glutamyl peptides have been identified in Spanish dry-cured hams yet. In this research, nine γ-glutamyl dipeptides (γ-EA, γ-EC, γ-EE, γ-EF, γ-EL, γ-EM, γ-EV, γ-EW, and γ-EY) and two γ-glutamyl tripeptides (GSH and γ-EVG) have been quantitated at 6, 12, 18 and 24 months of traditional processing of Spanish dry-cured ham by performing a Q Exactive Orbitrap-based tandem mass spectrometry. The results show an increase of γ-EA, γ-EE, γ-EF, γ-EL, γ-EM and γ-EVG, obtaining maximums at 24 months of curing ranging from 0.14 (γ-EVG) to 18.86 (γ-EL) μg/g dry-cured ham. Otherwise, γ-EV, γ-EW and γ-EY accumulated until the 18th month of storage to 15.10, 0.54 and 3.17 μg/g dry-cured ham, respectively; whereas γ-EC and GSH amounts decreased starting from 0.0676 and 4.41 μg/g dry-cured ham, respectively at earlier stages. The concentration dynamics of these compounds may be linked with proteolytic and oxidative reactions during processing. In addition, due to their synergistic effect on kokumi activity, this could constitute insights of the brothy perceptions of dry-cured ham, and these peptides probably contribute to the sensory differences existing in long processed Spanish dry-cured hams.

A Dual Stimuli-Responsive Nanoplatform Loaded PtIV -Triptolide Prodrug for Achieving Synergistic Therapy toward Breast Cancer.

To strengthen the antitumor efficacy and avoid toxicity to normal cells of cisplatin and triptolide,herein, an acid and glutathione (GSH) dual-controlled nanoplatform for enhanced cancer treatment through the synergy of both "1+1" apoptosis and "1+1" ferroptosis is designed. Remarkably,ZIF8in response totumor microenvironmentenhances drug targeting and protects drugs from premature degradation. Meanwhile, the PtIV center can be easily reducedtocisplatin because of the large amount of GSH, thus liberating the triptolide as the coordinated ligand. The releasedcisplatin and hemin in turn boost the tumor cell "1+1" apoptosis through chemotherapy and photodynamic therapy, respectively. Furthermore, GSH reduction throughPtIV weakens the activation of glutathione peroxidase 4 (GPX4) effectively. The released triptolide can inhibit the expressions of GSH by regulating nuclear factor E2 related factor 2 (Nrf2), further promoting membrane lipid peroxidation, thus "1+1"ferroptosis can be achieved. Bothinvitroandinvivoresults demonstrate that the nanosystem can not only perform superior specificity and therapeutic outcomes but also reducethe toxicity to normal cells/tissues of cisplatin and triptolide effectively. Overall, the prodrug-based smart system provides an efficient therapeutic strategy for cancer treatment by virtue of the effect of enhanced "1+1" apoptosis and "1+1" ferroptosis therapies.

Fluoxetine induces oxidative stress-dependent DNA damage in human hepatoma cells

<abstract> <p>Fluoxetine is a selective serotonin reuptake inhibitor that is a commonly used drug for the treatment of depression and obsessive-compulsive disorders. Despite the positive effects of this drug, it seems to be associated with various side effects. Genotoxicity or DNA damage is an important side effect of some kinds of drugs. To date, the genotoxicity and cytotoxicity of fluoxetine are partially unknown. In the present study, some oxidative stress methods were used, such as ROS, MDA and GSH evaluation methods in HepG2 cells treated with fluoxetine (1–10 µM). A comet assay was used to evaluate the genotoxic effects of fluoxetine, and flow cytometry was used for apoptosis detection in these hepatic cells. Our data have shown that fluoxetine increased MDA and intracellular concentration of ROS significantly (P < 0.001), while the amount of GSH was reduced significantly (P < 0.001). Our results also indicated that fluoxetine increased the DNA damage of HepG2 cells. The tail percentage of DNA for control cells was 4%, but this percentage was 19%, 28% and 32% for 1, 5 and 10 µM of fluoxetine concentration, respectively (P < 0.01 and P < 0.001). The flow cytometry results have also shown increases in early and late apoptosis for fluoxetine (13.31% and 9.54%, respectively). In conclusion, the present study has shown that fluoxetine is able to induce oxidative stress-dependent DNA damage. Anyway, more studies are needed to accurately explore the molecular and cellular aspects of fluoxetine.</p> </abstract>

Double protein directed synthesis of chemically etched sulfur doped quantum dots for signal "on-off-on" sensing of glutathione mediated by copper ions.

In this study, a novel "on-off-on" fluorescent probe was suggested for sensitive and selective assay of glutathione (GSH). The as-fabricated nanoswitch employs a Cu2+-sulfur quantum dot system (SQ-dots/Cu2+). The surface reactivity and water solubility of SQ-dots were improved through capping with egg white and bovine serum albumin proteins. The surface functional groups on the surface of double protein-protected SQ-dots enhanced the interaction with Cu2+ ions, resulting in the aggregation induced quenching of SQ-dots. Addition of GSH, a strong Cu2+ ion chelator, disassembles the large aggregates into relatively smaller ones, restoring the fluorescence emission of SQ-dots. Under optimized conditions, the fluorescence intensity was increased by increasing GSH amounts within the range of 0.13-550 μM with a detection limit (S/N = 3) of 0.04 μM. The SQ-dots/Cu2+ system was successfully applied for the detection of GSH in different matrices such as dietary supplements, human serum, and vegetable extract samples. The as-fabricated probe holds great potential for the synthesis of other functionalized SQ-dots for (bio) sensing.

Kadmiyuma Maruz Bırakılan Farelerin Oksidatif Stres Parametreleri ve Kan Lipid Profili üzerinde Resveratrolun Dokuya Özgü İyileştirici Etkisi

Cadmium can cause oxidative damage in organisms through overproduction of free radicals and suppression of the antioxidant defense system. Resveratrol is known as a nutraceutical with antioxidant properties accepted to have protective effects to the toxicity of heavy metals. In this study, we investigated if resveratrol could overwhelm the toxic effect of sub-acute cadmium exposure. Animals were divided to the following groups: 1) Control, 2) Vehicle control, 3) Cadmium (Cd), 4) Resveratrol (Res), 5) Exposure to both Cd and Res (Cd+Res). Increased lipid peroxidation and total sialic acids were determined in testis and lung tissues of mice exposed to Cd with the decreased in paraoxonase (PON1) level and GSH amount. Interestingly, we also found an increased lipid peroxidation and NO levels in lung tissue of mice exposed to Res and Cd + Res, but not in testis tissue. Moreover, increased triglycerides, total cholesterol, very low-density lipoproteins, and low-density lipoproteins as well as reduced high-density lipoproteins were found in mice exposed to Cd and Cd + Res compared to controls. Our results revealed that cadmium induces oxidative damage in tissues, via increased lipid peroxidation, total sialic acids and decreased antioxidant levels as well as disturbs lipid profile. Moreover, our findings indicate that resveratrol may act as “ameliorative” factor against the cadmium exposure in only testis tissue. Thus, we may suggest that ameliorative effect of resveratrol may vary depending on the exposure dose, exposure duration and exposed tissues of animals to the heavy metals.

The Effect of Thyme Essential Oil on Liver Injuries Caused by Renal Ischemia-Reperfusion in Rats.

Liver damage occurs following renal ischemia-reperfusion (RIR) that can cause inflammation and inflammatory cytokines activated after kidney injury. In this study, thyme essential oil (TE) with antioxidant and anti-inflammatory properties was used to reduce liver damage induced by renal IR. 32 male rats were randomly divided into 4 equal groups: (1) control, (2) RIR, (3) RIR+TE, and (4) TE. Rats received TE as a pretreatment at a dose of 0.5 ml/kg for one week. Then, under anesthesia for 45 minutes for ischemia, the kidneys of the animals were closed with clamps, and reperfusion was performed for 24 hours. Animal serum was isolated to evaluate alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) parameters. The liver of rats was examined for the measurement of malondialdehyde (MDA), nitric oxide (NO), glutathione (GSH), glutathione peroxidase (GPX), catalase (CAT), and expression of genes such as interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and caspase-3. ALP, AST, ALT, MDA, NO, IL-6, TNF-α, and caspase-3 increased significantly in the RIR group compared to the control group (p < 0.05). GSH, GPX, and CAT decreased significantly in the RIR group compared to the control group (p < 0.05). TE caused a decrease in ALP, AST, ALT, MDA, NO, IL-6, and TNF-α compared to the RIR group and caused an increase in the amount of GSH, GPX, and CAT in the RIR group (p < 0.05). This study showed that TE has antioxidant and anti-inflammatory properties that reduce liver damage induced by RIR.

P042: Metabolic regulation of adaptive response to arginine deprivation in Hodgkin Lymphoma

Our group has previously shown an altered aminoacidic homeostasis in the microenvironment of classic Hodgkin`s Lymphoma (HL), due to arginine deprivation, which plays a major role in the immune-escape mechanisms and T-cell anergy induction. To study the metabolic adaptive response to arginine deprivation in vitro, we cultured three HL human cell lines (L428, L540, KMH2) with customized complete media or lacking Arg (R0), supplemented with 10% dialyzed fetal bovine serum, in six independent experiments to collect their global metabolomic analysis by gas chromatography-mass spectrometry (GC/MS) and liquid chromatography-tandem mass spectrometry (LC/MS/MS) platforms by Metabolon Inc and transcriptome profiling by RNA-seq. Apotpostosis and mitochondrial depolarization were measured by FACS. Findings were validated by qRT-PCR and Western Blot analysis. Gene set enrichment analysis (GSEA) showed deep transcriptome rearrangements in KMH2 and HDMYZ cell lines, involving upregulation of genes required for the unfolded protein response, p53 pathway and networks and proteosome degradation, with a minimal effect on metabolism features, except for the genes involved in lactate metabolism. Arg deprivation caused mitochondrial distress and transcriptional reprogramming, via induction of oxidative stress, affecting the mitochondrial activity, switching from glucose-based metabolism to mitochondrial oxidation of fatty acids, requiring the transfer of fatty acids from lipid droplets to mitochondria. RNA-seq analysis showed that induction of ferroptosis key genes GPX4, TXNRD1 and ACSL4 together with reduced intracellular GSH levels and increased amount of cystine and methionine sulfoxide occurred upon arginine deprivation. The low-energy metabolic state induced by the adaptive response to arg-deprivation posed KMH2-HL cells into a quiescence state, with elevated HMOX1 and HMOX-2 to scavenge the excess ROS with subsequent genome instability as shown by increased γHA2X+/ATM+ cells. In HDMYZ, the increased oxidative stress due to arginine deprivation induced the engagement of the UFMylation pathway. These findings were confirmed in vivo, since in peripheral blood of HL patients we found reduced amount of GSH, glutamine and S-adenosylhomocysteine, citrate, methionine (p<0.001), as detected by HPLC. Taken together, our data suggest how arginine deprivation can regulate lipid trafficking, ferroptosis and UFMylation, novel potential targets to overcome drug resistance in cHL.