Total Glutathione Content Research Articles

Mitigating aluminum chloride-induced toxicity in Drosophila melanogaster with peptide fractions from Euphorbia species

This study assessed the antioxidative and protective effects of peptide extracts from selected Euphorbia species on Aluminum Chloride (AlCl3)-induced toxicity in Drosophila melanogaster. Euphorbia humifusa (EHU), Euphorbia hirta (EHI), and Euphorbia graminae (EHG) were screened for bioactive peptides. The crude peptide extract and partially purified peptide fractions of all the plants were subjected to preliminary antioxidants activities through 2, 2-diphenyl-1-picrylhyhdrazyl (DPPH), and nitric oxide (NO) scavenging activities. The most active peptide fraction was subjected to biochemical studies using Drosophila melanogaster. Flies were treated with AlCl3 (100 mg/kg diet), peptide fraction (5 and 10 mg/kg diet), and cotreatment of AlCl3 and the fraction, respectively. After treatment, flies were homogenized for the determination of total thiol and Glutathione (non-protein thiol) content, catalase and Glutathione-S-transferase activities, and nitric oxide (nitrite/nitrate) and hydroperoxide levels. The antioxidant screening revealed that the peptide fraction from Euphorbia humifusa (PEHU) was the most significant compared to the control (ascorbic acid). The PEHU (5 and 10 mg/kg diet) maintained the redox status of the flies in the biochemical study. The PEHU significantly counteracted AlCl3-induced reduction in antioxidants (catalase, GST, GSH and Total thiol), increased nitric oxide levels, and acetylcholinesterase activity and prevented behavioral deficits flies. Hence, the peptide fraction of Euphorbia humifusa may shield against the life-threatening effects of free radicals associated with aluminum chloride toxicity.

Oxidative state is associated with migration distance, but not traits linked to flight energetics

Flight can be highly energy demanding, but its efficiency depends largely on flight style, wing shape and wing loading, and a range of morphological and lifestyle adaptations that can modify the cost of sustained flight. Such behavioural and morphological adaptations can also influence the physiological costs associated with migration. For instance, during intense flight and catabolism of reserves, lipid damage induced by pro‐oxidants increases, and to keep oxidative physiological homeostasis under control, the antioxidant machinery can be upregulated. Studies on the oxidative physiology of endurance flight have produced contradictory results, making generalization difficult, especially because multispecies studies are missing. Therefore, to explore the oxidative cost of flight and migration, we used samples collected during the breeding season from 113 European bird species and explored the associations of measures of antioxidant capacity (total antioxidant status, uric acid and glutathione concentration) and oxidative damage of lipids (malondialdehyde) with variables reflecting flight energetics (year‐round or specifically during migration) using a phylogenetic framework. We found that none of the traits predicting year‐round flight energy expenditure (flight style, wing morphology and flight muscle morphology) explained any measures of oxidative state. Our results suggest that birds endure their everyday flight exercise without or with low oxidative cost. However, oxidative damage to lipids and one component of the endogenous antioxidant system (uric acid), measured after the end of spring migration on breeding adult birds, increased with migration distance. Our results suggest that migration could have oxidative consequences that might be carried over to subsequent life‐history stages (breeding).

Electroacupuncture improves retinal function in myopia Guinea pigs probably via inhibition of the RhoA/ROCK2 signaling pathway

ObjectiveTo investigate the effect of electroacupuncture (EA) on retinal function in guinea pigs with negative lens-induced myopia (LIM) by inhibiting the RhoA/ROCK2 signaling pathway. MethodsGuinea pigs were randomly divided into normal control (NC) group, LIM group, EA group, SHAM acupoint (SHAM) group, and electro-acupuncture + ROCK pathway inhibitor Y27632 (EA + Y27632) group. The refraction, axial length, retinal blood flow density, choroidal vascular index, retinal physiological function, the contents of total antioxidant capacity (T-AOC), catalase (CAT), glutathione (GSH), superoxide dismutase (SOD) and malondialdehyde (MDA) of each group were determined. The changes in retinal tissue structure were observed by hematoxylin and eosin (H&E) staining, and the expression of the RhoA/ROCK2 signaling pathway-related molecules in the retina was measured by real-time quantitative polymerase chain reaction (qPCR) and Western blot. ResultsMyopic refraction, AL, and MDA content in the LIM and SHAM groups were significantly increased, retinal blood flow density and CVI, SOD, GSH, CAT, T-AOC content were decreased. After EA intervention, myopic refraction, AL, and MDA content decreased, retinal blood flow density and CVI, SOD, GSH, CAT, T-AOC content were increased. H&E staining showed that the thickness of the guinea pig retina, the thickness of the inner and outer layers of the nucleus, and the number of cells were significantly increased after EA intervention. qPCR and western blot analyses showed that the expression of RhoA、ROCK2、MLC、CollagenⅠ、MMP-2、TIMP-2 and α-SMA were elevated in the LIM and SHAM group than those in the NC group. Compared with the LIM group, the expression of EA group was significantly decreased. ConclusionsElectroacupuncture can improve retinal function by improving retinal blood flow, reducing retinal oxidative damage, inhibiting RhoA/ROCK2 signaling pathway and controlling extracellular matrix remodeling, thus delaying the occurrence and development of myopia.

Noninvasive assessment of liver fibrosis in mini pigs using an 18F–AlF-NOTA-RGD2 PET/CT molecular probe

To evaluate the efficacy of the 18F–AlF-NOTA-RGD2 positron emission tomography (PET)/computed tomography (CT) molecular probe for the noninvasive staging of liver fibrosis in mini pigs, a potential alternative to invasive diagnostic methods was revealed. This study used 18F–AlF-NOTA-RGD2 PET/CT imaging of mini pigs to assess liver fibrosis. The methods included synthesis and quality control of the molecular probe, establishment of an animal model of liver fibrosis, blood serum enzymatic tests, histopathological examination, PET/CT imaging, collagen content and expression, and mitochondrial reserve function assessment. The 18F–AlF-NOTA-RGD2 PET/CT molecular probe effectively differentiated various stages of liver fibrosis in mini pigs. Blood serum enzymatic tests revealed distinct stages of liver fibrosis, revealing significant increases in AST, ALT, TBIL, and DBIL levels as fibrosis advanced. Notably, ALT levels increased markedly in severe fibrosis patients. A gradual increase in collagen deposition and increasing α-SMA RNA expression and protein levels effectively differentiated between mild and severe fibrosis stages. Pathological examinations and Sirius Red staining confirmed these findings, highlighting substantial increases in collagen accumulation. PET/CT imaging results aligned with histopathological findings, showing that increased radiotracer uptake correlated with fibrosis severity. Assessments of mitochondrial function revealed a decrease in total liver glutathione content and mitochondrial reserve capacity, especially in patients with severe fibrosis. The 18F–AlF-NOTA-RGD2 PET/CT molecular probe is a promising tool for the noninvasive assessment of liver fibrosis, offering potential benefits over traditional diagnostic methods in hepatology.

STAT3 Regulates the Redox Profile in MDA-MB-231 Breast Cancer Cells.

Unbalanced redox status and constitutive STAT3 activation are related to several aspects of tumor biology and poor prognosis, including metastasis and drug resistance. The triple-negative breast cancer (TNBC) is listed as the most aggressive and exhibits the worst prognosis among the breast cancer subtypes. Although the mechanism of reactive oxygen species (ROS) generation led to STAT3 activation is described, there is no data concerning the STAT3 influence on redox homeostasis in TNBC. To address the role of STAT3 signaling in redox balance, we inhibited STAT3 in TNBC cells and investigated its impact on total ROS levels, contents of hydroperoxides, nitric oxide (NO), and total glutathione (GSH), as well as the expression levels of 3-nitrotyrosine (3NT), nuclear factor (erythroid-derived 2)-like 2 (Nrf2), and nuclear factor kappa B (NF-κB)/p65. Our results indicate that ROS levels depend on the STAT3 activation, while the hydroperoxide level remained unchanged, and NO and 3NT expression increased. Furthermore, GSH levels, Nrf2, and NF-κB/p65 protein levels are decreased in the STAT3-inhibited cells. Accordingly, TNBC patients' data from TCGA demonstrated that both STAT3 mRNA levels and STAT3 signature are correlated to NF-κB/p65 and Nrf2 signatures. Our findings implicate STAT3 in controlling redox balance and regulating redox-related genes' expression in triple-negative breast cancer.

Effects of Drought at Anthesis on Flag Leaf Physiology and Gene Expression in Diverse Wheat (Triticum aestivum L.) Genotypes

The current study aimed to quantify the effects of two drought intensities achieved by deprivation of watering for 45 and 65% of the volumetric soil moisture content (VSMC) for 14 days after wheat anthesis to identify physio-biochemical and molecular changes associated with drought tolerance in six genotypes with different drought tolerance. Drought at 65% of VSMC induced a significant decrease in the chlorophyll a content in the drought-sensitive genotype, which indicated a strong loss of photosynthetic reaction centres. Further, in the drought-tolerant genotype, the content of carotenoids, which could play a vital role in resisting water shortage stress, tended to increase. The increased production of malondialdehyde showed that the antioxidant system in the drought-sensitive genotypes was not properly activated. A significant decrease in catalase (CAT; EC 1.11.1.6) was observed at a 45% reduction in VSMC, compared to the control, in the drought-sensitive genotype, and at a reduction in VSMC of 65%, in all medium sensitive genotypes. Further, the drought-tolerant and -medium tolerant genotypes responded to drought with a decline in total glutathione concentrations with the intention to reinforce their defence system. Thereby, dehydroascorbate reductase (DHAR; EC 1.8.5.1), monodehydroascorbate reductase (MDHAR; EC 1.6.5.4), and glutathione reductase (GR; EC 1.6.4.2) were critical enzymes involved in the ascorbate–glutathione cycle together with CAT, showing their main role in the detoxification of ROS produced with the reduction in VSMC by 65%. The results of gene expression analysis showed that severe drought increased the levels of the DHN5 and WZY2 genes (that were significantly positively correlated) in the drought-tolerant genotype, whose grain weight, area, and length did not change in maturity. Also, it was seen that DHN5 expression showed a significant positive correlation with grain length and proline content at a 45% reduction in VSMC. The identification of different mechanisms under drought can contribute to the selection of drought-tolerant genotypes.

Characterization of biologically synthesized copper nanoparticles by Aspergillus carbonarius and evaluation of their activity in reducing the pathogenic stress of tomatoes infected with Fusarium oxysporum

This study was conducted to evaluate the efficiency of the fungus Aspergillus carbonarius in the synthesis of copper nanoparticles (CuNPs) from copper chloride solution, with the characterization of these nanoparticles and the demonstration of their effectiveness in inhibiting the pathogenic fungus Fusarium oxysporum causing tomato root rot disease. The results indicated that the treatment of copper chloride solution at 150 mM with A. carbonarius filtrate led to the production of CuNPs which characterized by a color change from blue to green, with the highest absorbance observed at 300 nm. X-ray diffraction (XRD) results revealed the crystalline structure with peaks at angles 28.47286, 47.43258, and 56.20644 ◦ corresponding to (111), (220), and (311) planes, respectively, matching the JCPDS card number 01-081-1841. Scanning electron microscopy images showed cubic, pyramidal, and prismatic shapes with dimensions ranging from 15 - 84.291 nm. In laboratory experiment, CuNPs inhibited the growth of the pathogenic fungus, with an inhibition zone of 1.6 cm, compared to copper chloride which recorded 0.9 cm. In the field, , the treatment of CuNPs led to an increase in the stress resistance indicators, reaching the highest values for CuNPs when combined with the fungicide Medazim in the American tomato cultivar. The concentrations of total phenols and glutathione in this treatment were 2.56 mg/g and 0.322 µg/g, respectively. The lowest concentration of proline was recorded as 4.31 µg/g in the treatment involving CuNPs against the fungal infection. CuNPs with the fungicide medazim, significantly outperformed other treatments, registering lowest disease severity at 12.22% in the American cultivar, compared to the highest disease severity was recorded in the treatment with the pathogenic fungus alone, reaching 91.16% in the Dutch cultivar. CuNPs significantly increased tomato fruit weight, both in healthy and infected plants. The American, Turkish, and Dutch tomato cultivars infected with the pathogenic fungus and treated with under + medazim showed higher productivity, reaching 6432.55, 6334.34, and 6305.73 g.plant-1, compared to the lowest productivity in the treatment of these cultivars with the pathogenic fungus, reaching 1008.34, 977.45, and 833.78 g.plant-1, respectively

Pulsed Electric Field Technology for the Extraction of Glutathione from Saccharomyces cerevisiae.

Glutathione is a potent antioxidant that has shown promise in enhancing the processing of various foods and drinks such as bread and wine. Saccharomyces cerevisiae stands as a primary microorganism for glutathione production. This study sought to assess the potential of pulsed electric fields (PEFs) in extracting glutathione from S. cerevisiae cells. Yeast cells were subjected to PEF treatment (12 kV/cm, 150 µs) followed by incubation at varying pH values (4.0, 6.0, and 8.0) and temperatures (4 °C and 25 °C). Glutathione and protein extraction were assessed at different incubation times. Within one hour of incubation, PEF-treated yeast cells released over 60% of their total glutathione content, irrespective of pH and temperature. Notably, the antioxidant activity of the resulting extract surpassed that obtained through complete mechanical cell destruction and hot water, which form the conventional industrial extraction method in the glutathione industry. These results suggest that PEF could offer a rapid and more selective procedure, improving the extraction of this bioactive compound.

Carnitine modulates antioxidative defense in ABI2 mutant under salt stress

Carnitine, a ubiquitous compound in living organisms, fulfills diverse roles in energy metabolism, stress resilience, and detoxification. Its antioxidant and osmolyte traits offer relief to stressed plants. Antagonizing abscisic acid (ABA), carnitine influences ABA-responsive genes. Our study, using Arabidopsis thaliana wild-type Ler. (Landsberg erecta) and ABA-insensitive abi2-1 mutants, explored carnitine’s impact on antioxidative responses and ABI2’s role in salt-induced carnitine metabolism. The application of 5 µM carnitine has alleviated the decrease in RWC, shoot weight, and rosette diameter WT plants caused by 80 mM salt stress for 4 days. Carnitine reduced cell membrane damage and salinity effects, evidenced by decreased lipid peroxidation and H2O2. In contrast, the impaired ABI2 of abi2-1, due to deficient phosphatase activity, further exacerbated the inhibitory effect of carnitine on the enzymes of the ascorbate-glutathione cycle, consequently reducing stress mitigation. While abi2-1 mutants exhibited unchanged superoxide dismutase (SOD) activity, they demonstrated increased catalase and peroxidase activity following carnitine treatment under salt stress compared to WT plants. Conversely, wild-type WT plants treated with carnitine exhibited elevated total glutathione content under salt stress, a response not observed in abi2-1 mutants under carnitine treatment. These results underscore the crucial role of ABI2-dependent ABA signaling in regulating plant carnitine metabolism.

Dietary fatty acids modulate oxidative stress response to air pollution but not to infection.

Anthropogenic changes to the environment expose wildlife to many pollutants. Among these, tropospheric ozone is of global concern and a highly potent pro-oxidant. In addition, human activities include several other implications for wildlife, e.g., changed food availability and changed distribution of pathogens in cities. These co-occurring habitat changes may interact, thereby modulating the physiological responses and costs related to anthropogenic change. For instance, many food items associated with humans (e.g., food waste and feeders for wild birds) contain relatively more ω6-than ω3-polyunsaturated fatty acids (PUFAs). Metabolites derived from ω6-PUFAs can enhance inflammation and oxidative stress towards a stimulus, whereas the opposite response is linked to ω3-derived metabolites. Hence, we hypothesized that differential intake of ω6-and ω3-PUFAs modulates the oxidative stress state of birds and thereby affects the responses towards pro-oxidants. To test this, we manipulated dietary ω6:ω3 ratios and ozone levels in a full-factorial experiment using captive zebra finches (Taeniopygia guttata). Additionally, we simulated an infection, thereby also triggering the immune system's adaptive pro-oxidant release (i.e., oxidative burst), by injecting lipopolysaccharide. Under normal air conditions, the ω3-diet birds had a lower antioxidant ratio (GSH/GSSG ratio) compared to the ω6-diet birds. When exposed to ozone, however, the diet effect disappeared. Instead, ozone exposure overall reduced the total concentration of the key antioxidant glutathione (tGSH). Moreover, the birds on the ω6-rich diet had an overall higher antioxidant capacity (OXY) compared to birds fed a ω3-rich diet. Interestingly, only the immune challenge increased oxidative damage, suggesting the oxidative burst of the immune system overrides the other pro-oxidative processes, including diet. Taken together, our results show that ozone, dietary PUFAs, and infection all affect the redox-system, but in different ways, suggesting that the underlying responses are decoupled despite that they all increase pro-oxidant exposure or generation. Despite lack of apparent cumulative effect in the independent biomarkers, the combined single effects could together reduce overall cellular functioning and efficiency over time in wild birds exposed to pathogens, ozone, and anthropogenic food sources.

Foliar Application of Moringa Seed Extract Alone or in Combination with Salicylic Acid Enhanced Growth, Bioactive, and Phytohormone Compositions of Cancer Bush Plants under Heat Stress

Cancer bush (Lessertia frutescens L.) is an important medicinal plant that is rich in health beneficial compounds. It is commonly used in traditional medicine and as an ornamental plant. Heat stress is the most threatening abiotic factor restricting plant growth, thus causing crop yield and economic losses worldwide. The application of plant-derived biostimulant is as an innovative and promising approach for improving plant growth and productivity. The study was aimed to investigate the effect of moringa (Moringa oleifera Lam.) seed extract (MSE; 5%) either alone or in combination with salicylic acid (SA; 40 mg/L) on the growth, bioactive, and phytohormone attributes of cancer plants subjected to heat stress (38 °C for 2 hours for 5 days). Plants that were not treated were used as control. Plant pots were arranged in a randomized complete block design (RCBD) for treatments (MSE, SA, and MSE + SA) at 7-day intervals during the experiment. Both MSE and MSE + SA foliar application effectively increased plant growth characteristics and total carotenoids contents, and reduced electrolyte leakage and had no symptoms of wilting compared with SA and control. Plants treated with MSE showed higher number of branches and concentrations of abscisic acid (ABA), jasmonic acid (JA), and indole-3-acetic acid (IAA), and lower superoxide and hydrogen peroxide compared with other treatments and control. Also, plants treated with MSE + SA showed higher total chlorophylls and glutathione concentrations compared with other treatments and control. Overall, the application of MSE either alone or in combination with SA enhanced plant growth and productivity of heat-stressed cancer bush plants.

Estimating the synaptic density deficit in Alzheimer's disease using multi-contrast CEST imaging.

In vivo noninvasive imaging of neurometabolites is crucial to improve our understanding of the underlying pathophysiological mechanism in neurodegenerative diseases. Abnormal changes in synaptic organization leading to synaptic degradation and neuronal loss is considered as one of the primary factors driving Alzheimer's disease pathology. Magnetic resonance based molecular imaging techniques such as chemical exchange saturation transfer (CEST) and magnetic resonance spectroscopy (MRS) can provide neurometabolite specific information which may relate to underlying pathological and compensatory mechanisms. In this study, CEST and short echo time single voxel MRS was performed to evaluate the sensitivity of cerebral metabolites to beta-amyloid (Aβ) induced synaptic deficit in the hippocampus of a mouse model of Alzheimer's disease. The CEST based spectra (Z-spectra) were acquired on a 9.4 Tesla small animal MR imaging system with two radiofrequency (RF) saturation amplitudes (1.47 μT and 5.9 μT) to obtain creatine-weighted and glutamate-weighted CEST contrasts, respectively. Multi-pool Lorentzian fitting and quantitative T1 longitudinal relaxation maps were used to obtain metabolic specific apparent exchange-dependent relaxation (AREX) maps. Short echo time (TE = 12 ms) single voxel MRS was acquired to quantify multiple neurometabolites from the right hippocampus region. AREX contrasts and MRS based metabolite concentration levels were examined in the ARTE10 animal model for Alzheimer's disease and their wild type (WT) littermate counterparts (age = 10 months). Using MRS voxel as a region of interest, group-wise analysis showed significant reduction in Glu-AREX and Cr-AREX in ARTE10, compared to WT animals. The MRS based results in the ARTE10 mice showed significant decrease in glutamate (Glu) and glutamate-total creatine (Glu/tCr) ratio, compared to WT animals. The MRS results also showed significant increase in total creatine (tCr), phosphocreatine (PCr) and glutathione (GSH) concentration levels in ARTE10, compared to WT animals. In the same ROI, Glu-AREX and Cr-AREX demonstrated positive associations with Glu/tCr ratio. These results indicate the involvement of neurotransmitter metabolites and energy metabolism in Aβ-mediated synaptic degradation in the hippocampus region. The study also highlights the feasibility of CEST and MRS to identify and track multiple competing and compensatory mechanisms involved in heterogeneous pathophysiology of Alzheimer's disease in vivo.

Involvement of GLR-mediated nitric oxide effects on ROS metabolism in Arabidopsis plants under salt stress

Plant glutamate receptor-like channels (GLRs) play important roles in plant development, immune response, defense signaling and Nitric oxide (NO) production. However, their involvement in abiotic stress responses, particularly in regulating Reactive Oxygen Species (ROS), is not well understood. This study aimed to investigate GLR-mediated NO production on ROS regulation in salt-stressed cells. To achieve this, Arabidopsis thaliana Columbia (Col-0) were treated with NaCl, glutamate antagonists [(DNQX (6,7-dinitroquinoxaline-2,3-dione and AP-5(D-2-amino-5-phosphono pentanoic acid)], and NO scavenger [cPTIO (2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt)]. Salt-stressed plants in combination with DNQX and AP-5 have exhibited higher increase in lipid peroxidation (TBARS), hydrogen peroxide (H2O2) and superoxide radical (O−2) contents as compared to solely NaCl-treated plants. Furthermore, NO and total glutathione contents, and S-nitrosoglutathione reductase (GSNOR) activity decreased with these treatments. AP-5 and DNQX increased the activities of NADPH oxidase (NOX), catalase (CAT), peroxidase (POX), cell wall peroxidase (CWPOX) in salt-stressed Arabidopsis leaves. However, their activities (except NOX) were significantly inhibited by cPTIO. Conversely, the combination of NaCl and GLR antagonists, NO scavenger decreased the activities of ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione reductase (GR), dehydroascorbate reductase (DHAR) and monodehydroascorbate reductase (MDHAR) resulting in elevated GSSG levels, a low GSH/GSSG ratio, impaired ROS scavenging, excessive ROS accumulation and cell membrane damage. The findings of this study provide evidence that GLR-mediated NO plays a crucial role in improvement of the tolerance of Arabidopsis plants to salt-induced oxidative stress. It helps to maintain cellular redox homeostasis by reducing ROS accumulation and increasing the activity of SOD, GSNOR, and the ASC-GSH cycle enzymes.

Rats' testicular toxicity induced by bisphenol A is lessened by crocin via an antiapoptotic mechanism and bumped P-glycoprotein expression

Bisphenol A (BPA) engenders testicular toxicity via hydroxyl free radical genesis in rat striatum and depletion of the endogenous antioxidants in the epididymal sperms. The multi-drug resistance efflux carrier; P-glycoprotein (P-gp) expel the BPA from the testis and is responsible for the testicular protection through the deactivation of numerous xenobiotics. In our study, we investigated whether the BPA-induced testicular toxicity could be circumvented through administration of an antioxidant; crocin (Cr). Implication of P-gp expression was also investigated. Rats administered BPA (10 mg/kg b.w. orally for 14 days), dropped the body weight, testes/body weight ratio, total protein content, testosterone, follicle stimulating hormone, luteinizing hormone, and sperm motility & count, total antioxidant status, glutathione content and antioxidant enzymes (superoxide dismutase and catalase), concomitant with the elevation of the percentage abnormal sperm morphology, as well as testicular lipid peroxides and nitrite/nitrate levels. Histopathological examination showed spermatogenesis disorders after the BPA rats exposure. The immunohistochemical study showed up-regulation of the P-gp as evident by increasing immunoreactivity in interstitial cells, with positive localization in some spermatogonia cells. The BPA-treated rats showed positive immunoreactivity against caspase-3. The co-intake of Cr (200 mg/kg b.w./day, i.p. 14 days) along with the BPA, significantly ameliorated all the mentioned parameters, boosted histopathological image, fell the caspase-3 up-regulation, and perched the P-gp expression. We showed that, Cr promotes P-gp as an approach to nurture the testicles against the BPA toxicity. In conclusion; Cr lessens the oxidative stress conditions to safeguard rats from the BPA-induced testicular toxicity and sex hormones abnormalities, reducing apoptosis and up-regulating P-gp.

Amitriptyline ecotoxicity in Danio rerio (Hamilton, 1822) embryos – similar toxicity profile in the presence of nanoplastics

Interaction of nanoplastics (NPls) with other environmental contaminants could affect their uptake by the organisms and their toxicity. Thus, the present study aims to investigate the polystyrene NPls (44 nm) interaction with the antidepressant amitriptyline (AMI) and its toxicity to Danio rerio embryos. A similar toxicological profile for NPls + AMI exposure was found for most of the evaluated endpoints, comparing with AMI single exposure, showing that the presence of NPls did not modulate the AMI toxicity. However, the behavioral assessment showed a different pattern with hypoactivity for the NPls + AMI exposure (NPls – hyperactivity; AMI – no effect). Interaction effects between NPls and AMI were also found in the protein contents (antagonism) and in the total glutathione content (synergism). This study highlights the complexity and unpredictability of NPls interaction with pharmaceuticals, important for an accurate environmental risk assessment and for the developing of effective strategies and interventions against plastic pollution.

Loss of testosterone induces postprandial insulin resistance and increases the expression of the hepatic antioxidant flavin-containing monooxygenases in mice exposed to intermittent hypoxia.

We tested the hypothesis that low testosterone alters the effects of intermittent hypoxia (IH) on glucose homeostasis, hepatic oxidative stress, and transcriptomic profile in male mice. We used sham-operated or orchiectomized (ORX) mice exposed to normoxia (Nx) or IH for 2 weeks. We performed fasting insulin and glucose tolerance tests and assessed fasting and postprandial insulin resistance with the HOMA-IR. The activity of hepatic prooxidant (NADPH oxidase-NOX), antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase-SOD, Cat, GPx), lipid peroxidation (MDA concentration), and the total concentration of glutathione (GSH) were measured under postprandial conditions. mRNA sequencing and pathway enrichment analyses were used to identify hepatic genes underlying the interactions between IH and testosterone. In Sham mice, IH improves fasting insulin sensitivity and glucose tolerance, while there are no effects of IH in ORX mice. In ORX mice, IH induces postprandial hyperinsulinemia, insulin resistance, and a prooxidant profile of enzyme activity (low SOD activity) without altering hepatic MDA and GSH content. ORX and IH altered the expression of genes involved in oxidoreductase activities, cytochromes-dependent pathways, and glutathione metabolism. Among the genes upregulated in ORX-IH mice, the flavin-containing monooxygenases (FMO) are particularly relevant since these are potent hepatic antioxidants that could help prevent overt oxidative stress in ORX-IH mice. Low levels of testosterone in male mice exposed to IH induce post-prandial hyperinsulinemia and insulin resistance and determine the mechanisms by which the liver handles IH-induced oxidative stress.

Alkaloidal Extracts from Avicennia africana P. Beauv. (Avicenniaceae) Leaf: An Antiplasmodial, Antioxidant, and Erythrocyte Viable.

The emergence of drug-resistant parasites impedes disease management and eradication efforts. Hence, a reinvigorated attempt to search for potent lead compounds in the mangroves is imperative. This study evaluates in vitro antiplasmodial activity, antioxidant properties, and cytotoxicity of A. africana leaf alkaloidal extracts. The A. africana leaves were macerated with 70% ethanol to obtain a total crude extract. Dichloromethane and chloroform-isopropanol (3 : 1, v/v) were used to extract the crude alkaloids and quaternary alkaloids from the total crude. The antiplasmodial activities of the alkaloidal extracts were performed against 3D7 P. falciparum chloroquine-sensitive clone via the SYBR Green I fluorescence assay with artesunate serving as the reference drug. The alkaloidal extracts were further evaluated for antioxidant properties via the total antioxidant capacity (TAC), the total glutathione concentration (GSH), the DPPH (2,2-diphenyl-1-picrylhydrazyl) assay, and the ferric-reducing antioxidant power (FRAP) methods. The cytotoxic activity of the alkaloidal extracts was tested on erythrocytes using a 3-(4,5-dimethylthiazol-2-yl)-5-diphenyltetrazolium bromide-MTT assay with little modification. The phytocompounds in the alkaloidal extracts were identified via gas chromatography-mass spectrometry (GC-MS) techniques. The total crude extract showed good antiplasmodial activity (IC50 = 11.890 µg/mL). The crude and quaternary alkaloidal extracts demonstrated promising antiplasmodial effects with IC50 values of 6.217 and 6.285 µg/mL, respectively. The total crude and alkaloidal extracts showed good antioxidant properties with negligible cytotoxicity on erythrocytes with good selectivity indices. The GC-MS spectral analysis of crude alkaloidal extracts gave indole and isoquinoline alkaloids and several other compounds. Dexrazoxane was found to be the main compound predicted, with an 86% peak area in the quaternary alkaloidal extract. The crude and quaternary alkaloidal extracts exhibited antiplasmodial activities and ability to inhibit oxidative stress with negligible toxicity on erythrocytes. This may be good characteristics to avoid oxidative stress related to Plasmodium infection in the treatment of malaria.

Metodologias para indicar estresse causado por jejum em poedeiras comerciais

ABSTRACT: Plasma corticosterone concentration (CORT), heterophil:lymphocyte ratio (H:L), catalase activity (CAT), total glutathione concentration (GSH), and thiobarbituric acid reactive substance levels (TBARS) were evaluated in 48 Hy-Line Brown laying hens, at 79 weeks of age, after being fasted for 10 consecutive days. Blood was collected on days zero, two, four, six, eight, and 10 of fasting, and a completely randomized design was adopted, with eight replicates on each day of collection, with each bird being an experimental unit. The time of maximum stress was determined for each method, using the polynomial regression analysis. The Pearson correlation analysis was also performed to determine whether the methods were interchangeable. CORT and GSH concentrations indicated that the time of maximum stress occurred at 4.3 days of fasting, whereas, the H:L and CAT activity indicated that the time of maximum stress occurred at 10 days of fasting. The malondialdehyde concentration detected by the TBARS method was highest at day zero and lowest at day 5.5 of fasting, but this method was not a reliable measure of stress. The low Pearson correlation coefficients observed among the methods made it impossible to designate only one of the tested methods as a replacement for the others, to measure the stress in laying hens during fasting.

Potential involvement of ferroptosis in BPA-induced neurotoxicity: An in vitro study

BackgroundFerroptosis is a newly recognized cell death pathway having distinct characteristics compared to traditional cell death pathways such as apoptosis, necroptosis, or autophagy. However, the potential involvement of ferroptosis in bisphenol A (BPA)-induced neurotoxicity has not been well explored so far. In present study, we analyzed the relationship between ferroptosis and BPA-induced neurotoxicity. MethodsIn this study, a human neuroblastoma cell line, SH-SY5Y, was treated with BPA, ferrostatin-1 (FS-1, ferroptosis inhibitor) and RSL-3 (ferroptosis inducer). The cell viability was measured using MTT assay. Additionally, the levels of lipid peroxidation, total iron content, reactive oxygen species (ROS) generation, and nitrite content were measured to evaluate the key markers of ferroptosis. To further confirm the involvement of ferroptosis in BPA-induced neurotoxicity, other ferroptosis markers such as glutathione peroxidase (GPx) activity, total glutathione contents and antioxidant parameters were also evaluated. ResultsThe cell viability of SH-SY5Y cells was down-regulated by BPA treatment in a concentration-dependent manner, the cell viability at 0.1 µM concentration was 97.63% whereas at highest BPA concentration i.e. 10 µM, the cell viability was 86.05% (p < 0.0001). Also the antioxidant parameters including catalase and superoxide dismutase activity of neuronal cells were down-regulated upon BPA exposure. However, the levels of lipid peroxidation, total iron, reactive oxygen species, and nitrite contents were increased in a concentration-dependent manner which could be rescued by FS-1 and exacerbated by RSL-3. The total iron in SH-SY5Y cells at 0.1 µM concentration was found to be 1.2 fold (p < 0.05) of control and at highest BPA concentration total iron was about 1.41 fold (p < 0.001) of control. ConclusionsThe present study indicated that, ferroptosis plays an important role in the progression of BPA-induced neurotoxicity, and ferroptosis may become a novel target in the treatment of various neurological disorders.

Effects of dietary methionine supplementation on the growth performance, immune responses, antioxidant capacity, and subsequent development of layer chicks

Deficiencies or excesses of dietary amino acids, and especially of methionine (Met), in laying hens can lead to abnormal protein anabolism and oxidative stress, which affect methylation and cause cellular dysfunction. This study investigated the effects of dietary methionine (Met) levels on growth performance, metabolism, immune response, antioxidant capacity, and the subsequent development of laying hens. A total of 384 healthy 1-day-old Hyline Grey chicks of similar body weight were randomly allocated to be fed diets containing 0.31%, 0.38%, 0.43% (control group), or 0.54% Met for 6 wk, with 6 replicates of 16 chicks in each. The growth performance of the chicks was then followed until 20 wk old. The results showed dietary supplementation with 0.43% or 0.54% Met significantly increased their mean daily body weight gain, final weight, and Met intake. However, the feed:gain (F/G) decreased linearly with increasing Met supplementation, from 0.31 to 0.54% Met. Met supplementation increased the serum albumin, IgM, and total glutathione concentrations of 14-day-old chicks. In contrast, the serum alkaline phosphatase activity and hydroxyl radical concentration tended to decrease with increasing Met supplementation. In addition, the highest serum concentrations of IL-10, T-SOD, and GSH-PX were in the 0.54% Met-fed group. At 42 d of age, the serum ALB, IL-10, T-SOD, GSH-PX, T-AOC, and T-GSH were correlated with dietary Met levels. Finally, Met supplementation reduced the serum concentrations of ALP, IL-1β, IgA, IgG, hydrogen peroxide, and hydroxyl radicals. Thus, the inclusion of 0.43% or 0.54% Met in the diet helps chicks achieve superior performance during the brooding period and subsequently. In conclusion, Met doses of 0.43 to 0.54% could enhance the growth performance, protein utilization efficiency, antioxidant capacity, and immune responses of layer chicks, and to promote more desirable subsequent development during the brooding period.