Thiol Content Research Articles

Crocin Improves Cognitive Impairment in LPS-treated Rats through Anti-Apoptotic, Anti-Inflammatory, and Antioxidant Activities.

Brain inflammation and oxidative stress play critical roles in neuronal apoptosis and memory dysfunction in Alzheimer's disease. Crocin, a natural carotenoid in the stigma of saffron, possesses radical scavenging, anti-inflammatory, and anti-apoptotic properties. This study investigates the protective impact of crocin on neuronal apoptosis, oxidative stress, neuroinflammation, and memory deficits induced by lipopolysaccharide (LPS) in rats. Male Wistar rats received 100mg/kg of crocin for 12 days, with LPS (1mg/kg, ip) injected on days 8-12. Spatial learning and memory were evaluated in the Morris water maze two hours after LPS injection. Gene expression of nuclear factor kappa B (NF-κB), tumor necrosis factor-α (TNF-α), caspase 3, and lipid peroxidation was assessed in hippocampal homogenates at the end of the behavioral test. Histopathological changes in the hippocampus and cerebral cortex were evaluated using H&E staining. The results indicated that LPS administration caused spatial learning and memory dysfunction (P = 0.001, P < 0.01) accompanied by upregulation of Nfkb, Tnfα, and Casp3 mRNA expression (P < 0.0001), increased TNF-α (P < 0.01) and lipid peroxidation level (P < 0.01), decreased total thiol concentration (P < 0.05), tissue damage and neuronal loss in the hippocampus (P < 0.0001). Furthermore, crocin treatment at a dosage of 100mg/kg attenuated learning and memory impairments (P = 0.001, P < 0.01), downregulated Nfkb, Tnfα, and Casp3 mRNA expression (P < 0.0001), decreased TNF-α level (P < 0.01) and lipid peroxidation (P < 0.05) and increased total thiol level (P < 0.05) in the hippocampus. Crocin also ameliorated LPS-induced pathological changes and neuronal loss in the hippocampus (P < 0.001) and cerebral cortex (P < 0.01). In conclusion, the neuroprotective effects of crocin against LPS-induced histopathological and behavioral changes could be attributed to its anti-apoptotic, anti-inflammatory, and radical-scavenging activities in the rat brain.

Combination of Essential Oils and Extracts Enriched with Tocopherols in Diet of Holstein Steers and Its Effect on Ruminal Fermentation, Antioxidant and Anti-Inflammatory Response, Feed Efficiency and Fatty Acid Profile in Meat

Background: This experiment evaluated whether the combination of essential oils (thyme, rosemary, orange) with extracts of Cinnamomum and Quillaja saponaria (enriched with tocopherols) in steers’ diet has positive effects on the health, digestibility, growth, oxidative stress, and fatty acid profile of meat. Methods: The experiment included 24 Holstein steers, divided into two groups, randomly divided (treated = phytogenic mixture (n = 12); control = monensin (n = 12)), undergoing a growth phase of 120 days and 90 days of finishing in individual pens, with a monthly collection of blood, rumen fluid, feces, and feed. Results: In the finishing phase (days 150 to 210), there was a tendency (p = 0.07) toward more significant average daily weight gain when cattle consumed a phytogenic mixture than monensin. The use of the phytogenic product showed greater feed efficiency in the period from 150 to 240 days (p &lt; 0.05), with higher levels of short-chain fatty acids (SCFAs), without differences in the composition of these SCFAs between the control and treated groups, with a reduction in apparent digestibility in the treated group. In blood parameters, the treated group showed higher erythrocyte counts (p &lt; 0.05) without changes in leukocytes, lymphocytes, granulocytes, monocytes, and platelets. The phytogenic treatment demonstrated higher total protein and globulin levels, with increased glucose levels on days 30 and 210 (p &lt; 0.05). Ceruloplasmin and haptoglobin levels increased in the treated group without changes in ferritin, transferrin, C-reactive protein, IgA, and heavy-chain immunoglobulins (p &lt; 0.05). A reduction in oxidative stress was observed using the phytogenic agent, with a marked decrease in the levels of reactive oxygen species (ROS) and carbonyl proteins, with a tendency to increase glutathione S-transferase (p &lt; 0.05). In meat, the phytogenic reduced saturated fatty acids with increased polyunsaturated fatty acids and increased the omega 6/omega 3 ratio. Lower levels of thiobarbituric acid reactive substances and ROS were detected in the meat combined with a higher concentration of total thiols in the treated group than in the control group. Conclusions: The use of the phytogenic in steers’ diets has shown to be a viable candidate for replacing monensin, presenting equal or superior performance results with systemic modulating effects on oxidative stress, meat quality, and serum and biochemical parameters that contribute to more substantial health and efficiency.

Gallic acid attenuates lipopolysaccharide - induced memory deficits, neurochemical changes, and peripheral alterations in purinergic signaling.

Neuroinflammation is associated with many neurological disorders. Gallic acid (GA) has attracted significant attention due to its biological properties, such as neuroprotective, anti-inflammatory, and antioxidant effects. In this study, we evaluated the effects of GA in memory, TNF-α levels, oxidative stress, and activities of acetylcholinesterase (AChE), Na+, K+-ATPase and Ca2+-ATPase in the brain of mice exposed to lipopolysaccharide (LPS). Additionally, we evaluated alterations in adenine nucleotides and nucleosides in the serum. Male mice were orally pretreated with vehicle or GA (50 or 100mg/kg) for 14 days. Between days 8 and 14, the animals also received LPS injection (250µg/kg) or saline. At the end of the experimental protocol, the animals were submitted to object recognition test, euthanized and cerebral cortex, hippocampus, striatum and blood were collected. LPS induced memory deficits, which were prevented by GA treatment. GA protected against LPS-induced oxidative damage in the cerebral cortex, hippocampus and striatum by reducing reactive oxygen species and nitrite levels, while increasing total thiol content and activities of antioxidant enzymes. GA also prevented LPS-induced alterations in AChE, Na+, K+-ATPase, and Ca2+-ATPase activities in brain structures. LPS elevated TNF-α levels in the hippocampus and cerebral cortex, which were attenuated by GA treatment. Furthermore, LPS caused a reduction in ADP and AMP hydrolysis and an increase in adenosine deamination in the serum, which were also prevented by GA. The effects of GA against neuroinflammation may be attributed to its potent antioxidant and anti-inflammatory properties, which modulate various pathways, including those involved in memory mechanisms.

Changes in Biomarkers of Inflammation and Oxidative Status in Dogs Subjected to Celiotomy or Video-Assisted Ovariohysterectomy.

We evaluated the surgical stress response of dogs undergoing three ovariohysterectomy (OVH) techniques. Twenty-nine healthy females were allocated into groups: celiotomy and ligature (CelioSut), celiotomy and bipolar energy (CelioBip), and a video-assisted technique using two portals and bipolar energy (VidBip). Clinical evaluation was performed, and the following blood analyses were determined: acute phase proteins (C-reactive protein and haptoglobin), white blood cell counting (WBC), and biomarkers of oxidative status. The VidBip required more time despite a smaller incision, lower heart rate, and earlier feeding and urination. All groups had high white blood cells counts; the C-reactive protein (CRP) levels peaked at 6 and 12 h in all groups and was higher in the CelioBip and VidBip groups compared to CelioSut; haptoglobin concentrations peaked at 48 h in all groups and increased in dogs of the CelioSut group. Trolox equivalent antioxidant capacity, ferric reducing ability of plasma, cupric reducing antioxidant capacity, and advanced oxidation protein were not significantly different among the groups or time points. Total thiol concentrations were lower in CelioBip and CelioSut groups. All surgical techniques induced an inflammatory and oxidative stress response, but the video-assisted technique produced early clinical recovery. The bipolar device produces fewer disturbances than suspensory ligament rupture and ligature.

Effects of aging and repeated freeze-thaw cycles on quality attributes, physicochemical and biochemical properties, and sensory characteristics of beef sirloins

This study aimed to evaluate the influence of aging before freezing and repeated freezing on quality attributes of beef sirloins. At 5 days postmortem, pairs of sirloins (M. gluteus medius) were collected from 20 beef carcasses (USDA Choice grade). The sirloin samples from each side were assigned to either no additional aging (A0) or 4 weeks of aging (A4). Samples were then subjected to either single freeze-thawing (A0FT and A4FT) or repeated freeze-thawing (A0FT × 2 and A4FT × 2) over 3 weeks. Following aging (2 °C), freezing (−20 °C), and thawing (2 °C), the samples were evaluated for water-holding capacity (WHC), Warner-Bratzler shear force (WBSF), myofibrillar fragmentation index (MFI), color, 2-thiobarbituric acid reactive substances (TBARS), carbonyl and thiol contents, and consumer sensory evaluation (n = 120). Freezing without aging increased freeze-thaw and drip losses in A0FT and A0FT × 2 compared to A4FT and A4FT × 2 (p < 0.05). Freezing and repeated freezing decreased WBSF values (p < 0.05) regardless of aging status. Similarly, MFI was higher in A0FT and A4FT compared to A0 and A4, respectively (p < 0.05). Both freezing and repeated freezing increased TBARS levels (p < 0.05). Moreover, freezing without aging reduced CIE L*, a*, b* and chroma values (p < 0.05). However, aged/frozen samples retained similar color attributes to never frozen counterparts (p > 0.05). Consumer sensory panel reported improved sensory attributes of aged/frozen samples, without any adverse effects from repeated freezing. These findings indicate that aging can improve WHC, color, and tenderness of single or repeated frozen/thawed beef sirloins, suggesting it as a practical strategy for the industry to improve meat quality and value of frozen/thawed beef.

A Novel Concept for Cleavable Linkers Applicable to Conjugation Chemistry - Design, Synthesis and Characterization.

Linkers with disulfide bonds are the only cleavable linkers that utilize physiological thiol gradients as a trigger to initiate the intracellular drug release cascade. Herein, we present a novel concept exploiting the thiol gradient phenomena to design a new class of cleavable linker with no disulfide bond. To support the concept, an electron-deficient sulfonamide-based cleavable linker amenable to conjugation of drug molecules with targeting agents, was developed. Modulating the electron-withdrawing nature of the aryl sulfonamide was critical to the balance between the stability and drug release. Favorable stability and payload release in human serum under physiologically relevant thiol concentrations was demonstrated with two potent cytotoxics. Intracellular payload release was further validated in cell-based assay in context of antibody-drug conjugate generated from monoclonal antibody and sulfonamide containing linker. To support the proposed release mechanism, possible downstream by-products formed from the drug-linker adduct were characterized.

Darbepoetin alpha has an anxiolytic and anti-neuroinflammatory effect in male rats

AimsWe aimed to investigate the anxiolytic effect of darbepoetin alpha (DEPO), an erythropoietin derivative, in a neuroinflammation model regarding different behaviors and biological pathways.MethodsForty adult male Wistar albino rats were divided into four groups (control, LPS, DEPO, and DEPO + LPS). The rats were treated with 5 µg /kg DEPO once a week for four weeks, after which neuroinflammation was induced with 2 mg/kg lipopolysaccharide (LPS). The elevated plus maze, open-field, and light‒dark box tests were conducted to assess anxiety levels. Harderian gland secretions were scored via observation. Tumor necrosis factor alpha (TNF-α), Interleukin-1-beta (IL-1β), brain-derived growth factor (BDNF), serotonin, cortisol, total antioxidant/oxidant (TAS/TOS), and total/free thiol levels were measured in the prefrontal cortex, striatum, and serum.ResultsDEPO had a potent anxiolytic effect on both DEPO and DEPO + LPS groups. Compared to the control group, DEPO administration caused an increase in serotonin and BDNF levels and decreased basal cortisol and TNF-α levels in naive rats. IL-1β did not alter after DEPO administration in naive rats. Prophylactic DEPO treatment remarkably downregulated cortisol, IL-1β, and TNF-α in the DEPO + LPS group. In addition, prophylactic DEPO administration significantly attenuated the decrease in serotonin and BDNF levels in the DEPO + LPS group. Furthermore, DEPO ameliorated excessive harderian gland secretion in the DEPO + LPS group. Compared with those in the control group, the free thiol content in the serum increased after DEPO administration. No similar effect was seen in the DEPO + LPS group receiving prophylactic DEPO. TAS showed no difference among all experimental groups. DEPO administration increased TOS and OSI in the serum and prefrontal cortex but not in the striatum. This effect was not seen in the DEPO + LPS group.ConclusionDarbepoetin alpha had an anxiolytic effect on many physiological mechanisms in a neuroinflammation model and naive rats.

Low feed intake at weaning reduces intestinal glutathione levels and promotes cysteine oxidation to taurine in pigs.

Weaning stress in pigs is associated with low feed intake and poor nutrient utilization. Cysteine is a sulfur amino acid with key roles in pig production, but how cysteine metabolism and requirements are affected by weaning stress should be better defined. The objective of this study was to determine the collective impact of weaning and feed restriction on tissue cysteine metabolism. Pigs were weaned at 21-d age without access to feed (W; 6.90 ± 0.81kg; n = 9; reflecting acute nutritional stress) or were not weaned and remained with the sow (NW; 6.81 ± 0.65kg; n = 8). At euthanasia (23-d age), blood, bile, and liver, jejunum, and ileum tissues were collected. Plasma, bile, and tissue amino acid and amino thiol concentrations were analyzed by HPLC. Activity of glutamate cysteine ligase (GCL) and glutathione synthetase (GSS), enzymes needed for glutathione (GSH) production, and cysteine dioxygenase 1 (CDO1) were determined with activity assays followed by HPLC analysis of reaction products. Plasma (271 versus 192 ± 19 µmol/L; P < 0.001) and liver (417 versus 298 ± 33 nmol/g; P < 0.05) Cys concentrations were increased in W compared to NW pigs. Despite greater plasma Cys, jejunum and ileum Cys content were not affected by weaning (P > 0.10), whereas γ-glutamylcysteine (γ-GlyCys), the immediate precursor of GSH, declined in both jejunum (14.3 versus 9.7 ± 1.4 nmol/g; P < 0.01) and ileum (11.2 versus 6.4 ± 0.8 nmol/g; P < 0.001) in W pigs. Glutathione content was lower in the jejunum (1379 versus 1720 ± 70 nmol/g; P < 0.05) and ileum (1497 versus 1740 ± 74 nmol/g; P < 0.05) in W pigs. In the jejunum, GCL activity tended to be greater (0.56 versus 0.39 ± 0.07 nmol γ-GluCys • mg-1 • min-1; P < 0.10), whereas GSS activity tended to be lower (1.11 versus 1.38 ± 0.10 nmol GSH • mg-1 • min-1; P < 0.10) in W compared to NW pigs. In the ileum, the activities of GCL and GSS were not affected by weaning (P > 0.10). Although liver CDO1 activity was not different between groups (P < 0.10), liver taurine was greater in W compared to NW pigs (5115 versus 2336 ± 912 nmol/g; P = 0.001). Bile concentrations of Cys (1203 versus 279 ± 103 µmol/L; P < 0.001) and cysteinylglycine (203 versus 117 ± 33 µmol/L; P < 0.10), the direct product of GSH degradation, were greater in W compared to NW pigs. Collectively, these results suggest that systemic Cys is not effectively utilized for gut GSH production in newly weaned pigs and is instead oxidized to taurine and eliminated in bile.

OsbHLH6, a basic helix-loop-helix transcription factor, confers arsenic tolerance and root-to-shoot translocation in rice.

Arsenic (As) is extremely toxic to plants, posing a serious concern for food safety. Identification of genes responsive to As is significative for figuring out this issue. Here, we identified a bHLH transcription factor OsbHLH6 that was involved in mediating the processes of As tolerance, uptake, and root-to-shoot translocation in rice. The expression of OsbHLH6 gene was strongly induced after 3 and 48 h of arsenite [As(III)] treatment. The OsbHLH6-overexpressed transgenic rice (OE-OsbHLH6) was sensitive to, while the knockout mutant of OsbHLH6 gene (Osbhlh6) was tolerant to As(III) stress by affecting the contents of reactive oxygen species (ROS) and non-protein thiols (NPT), etc. Knockout of OsbHLH6 gene increased significantly the As concentration in roots, but decreased extensively As accumulation in shoots, compared to that in OE-OsbHLH6 and WT plants. The transcripts of phytochelatins (PCs) synthetase encoding genes OsPCS1 and OsPCS2, as well as As(III) transporter encoding genes OsLsi1 and OsABCC1 were greatly abundant in Osbhlh6 mutants than in OE-OsbHLH6 and WT plants under As(III) stress. In contrast, the expression of OsLsi2 gene was extensively suppressed by As(III) in Osbhlh6 mutants. OsbHLH6 acted as a transcriptional activator to bind directly to the promoter and regulate the expression of OsPrx2 gene that encodes a peroxidase precursor. Moreover, overexpression of OsbHLH6 gene resulted in significant change of expression ofamounts of abiotic stress-related genes, which might partially contribute to the As sensitivity of OE-OsbHLH6 plants. These findings may broaden our understanding of the molecular mechanism of OsbHLH6-mediated As response in rice and provide novel useful genes for rice As stress-resistant breeding.

Redox proteomic analysis of H2O2 -treated Jurkat cells and effects of bicarbonate and knockout of peroxiredoxins 1 and 2

Oxidation of thiol proteins and redox signaling occurs in cells exposed to H2O2 but mechanisms are unclear. We used redox proteomics to seek evidence of oxidation of specific proteins either by a mechanism involving reaction of H2O2 with CO2/bicarbonate to give the more reactive peroxymonocarbonate, or via a relay involving peroxiredoxins (Prdxs). Changes in oxidation state of specific Cys-SH residues on treating Jurkat T lymphoma cells with H2O2 were measured by isotopically labeling reduced thiols and analysis by mass spectrometry. The effects of bicarbonate and of knocking out either Prdx1 or Prdx2 were examined. Approximately 14,000 Cys-peptides were detected, of which ∼ 1% underwent 2-10 fold loss in thiol content with H2O2. Those showing the most oxidation were not affected by the presence of bicarbonate or knockout of either Prdx. Consistent with previous evidence that bicarbonate potentiates inactivation of glyceraldehyde-3-phosphate dehydrogenase, the GAPDH active site Cys residues were significantly more sensitive to H2O2 when bicarbonate was present. Several other proteins were identified as promising candidates for further investigation. Although we identified some potential protein candidates for Prdx- dependent oxidation, most of the significant differences between KO and WT cells were seen in proteins for which H2O2 unexpectedly increased their CysSH content over untreated cells. We conclude that facilitation of protein oxidation by bicarbonate or Prdx-mediated relays is restricted to a small number of proteins and is insufficient to explain the majority of the oxidation of cell thiols that occurs in response to H2O2.

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.

Evaluation of pregnancy toxemia in goats: Metabolic profile, hormonal findings, and redox balance

Pregnancy toxemia entails the development of hyperketonemia due to the negative energy balance that occurs in the last trimester of pregnancy in dairy goats. There are a limited number of studies on new diagnostic biomarkers and the pathophysiology of pregnancy toxemia in goats. This study assessed the hormones spexin and irisin and redox biomarker and metabolic profile changes in goats with pregnancy toxemia. The study included 36 hair goats, with 12 in the subclinical pregnancy toxemia (SPT) group, 12 in the clinical pregnancy toxemia (CPT) group, and 12 in the control group (CG). Spexin, irisin, and insulin hormone concentrations were determined by ELISA, while metabolic profile parameters were evaluated with an automatic chemical analyzer and redox balance by spectroscopy. In the goats with pregnancy toxemia, serum levels of spexin, irisin, insulin, and glucose were significantly lower than those of CG goats. The SPT group had significantly higher ischemia-modified albumin concentrations than CG goats, while the CPT group had significantly lower native thiol concentrations compared to CG animals. The concentrations of beta-hydroxybutyric acid and non-esterified fatty acids in the groups with pregnancy toxemia were significantly higher than those of control animals. In the diagnosis of pregnancy toxemia, spexin had sensitivity of 90 %, specificity of 95 %, and an area under the curve value of 0.988, while irisin had sensitivity of 90 %, specificity of 91 %, and an AUC value of 0.979. In this study, both spexin and irisin hormones showed excellent performance in the diagnosis of pregnancy toxemia. In addition, pregnancy toxemia was found to cause increased oxidative stress and significant changes in metabolic and endocrine metabolism in goats. Measuring these biomarkers in both healthy dairy goats and those with pregnancy toxemia may contribute significantly to the management and diagnosis of herd health.

Inhibitory effect of caffeic acid and dihydrocaffeic acid on the formation of advanced glycation end products: Mechanism analysis based on intermolecular interaction

The accumulation of advanced glycation end products (AGEs) generated from glycation reaction accelerates the development and deterioration of some diseases. Herein, the AGEs model of fructose-induced glycation of human serum albumin (HSA) was constructed and characterized. Both caffeic acid (CA) and dihydrocaffeic acid (DHCA) can inhibit the formation of AGEs, and the inhibitory effect of CA was more pronounced, as confirmed by fluorescence spectroscopy, gel electrophoresis, scanning electron microscopy, and carbonyl and thiol contents analysis. The binding of CA and DHCA to HSA was investigated by fluorescence spectroscopy and molecular docking. The primary inhibitory mechanism was the occupation of the glycation site LYS199 of HSA, and the more significant inhibitory effect of CA was related to its stronger binding affinity. Furthermore, the conformational changes caused by glycation and CA/DHCA binding were monitored using fluorescence, circular dichroism, and dynamic light scattering analysis. The results would help to elucidate the inhibitory mechanism of CA and DHCA on AGEs formation based on their different structures and inhibitory effects, and provide guidance for choosing natural inhibitors for diabetes.

Antioxidant effects of a novel pioglitazone analogue (PA9) in a rat model of diabetes: Modulation of redox homeostasis and preservation of tissue architecture

Oxygen-free radicals have been implicated in the initiation of diabetic complications. Thiazolidinediones (TZDs), known for their antidiabetic properties, also demonstrate notable antioxidant and anti-inflammatory effects. Although a recently developed imidazolyl analogue of pioglitazone (PA9) has exhibited superior glucose-lowering efficacy compared to pioglitazone, its antioxidant effects remain unexplored. Thus, the objective of this study is to evaluate the antioxidant properties of PA9 in animal models with diabetes.Rats were randomly separated into the following four groups: control, diabetic, and two groups treated orally with pioglitazone as a standard drug and PA9 for ten days. Upon completion of the experiment, tissues from the liver, heart, brain, pancreas, spleen, and kidneys were collected to assess oxidant/antioxidant markers and histological alterations. The administration of PA9 resulted in a noteworthy reduction in malondialdehyde (MDA) levels compared to the diabetic group (p < 0.05). The group receiving PA9 displayed elevated levels of three antioxidant markers, catalase (CAT), superoxide dismutase (SOD), and total thiol, in pancreatic tissue compared to diabetic rats (p < 0.05).Furthermore, increased content of CAT was evident in the heart (p < 0.05), spleen (p < 0.001), brain, and kidney tissues in the PA9-treated group, along with augmented thiol content in the spleen compared to the diabetic group. Remarkably, no significant histological changes were observed in the liver, pancreas, heart, brain, spleen, and kidneys of the PA9-treated groups relative to diabetic rats. PA9 effectively mitigates oxidative stress, modulates redox homeostasis, and shows promise in preventing diabetic complications. The proven safety profile of this analogue underscores its potential, warranting comprehensive clinical evaluation to thoroughly understand its therapeutic scope and efficacy in the management of diabetes.

Herniarin ameliorates acrylamide-induced neurotoxicity in rat: involvement of neuro-inflammation and acetylcholinesterase

This study aimed to investigate the protective effects of herniarin against acrylamide neuro-toxicity. Rats were administered 50 mg/kg of acrylamide (Acr) along with oral doses of herniarin at 50, 100, and 200 mg/kg for 11 days. After treatment, the animals were sacrificed and biochemical markers including superoxide dismutase (SOD), catalase activity, malondialdehyde (MDA), nitric oxide (NO), thiols and acetylcholine esterase (AChE) level were evaluated. Moreover, mRNA expression of neuro-inflammatory cytokines including TNF-α, IL-1β, and IL-6 were measured using qRT-PCR method. As results, Acr increased MDA with subsequent reduction in SOD, catalase, and thiols content. In contrast, administration of herniarin remarkably normalised the antioxidants and decreased lipid peroxidation. Furthermore, it downregulated mRNA expression of TNF-α, IL-1β, and IL-6 (p < 0.001) as well as decreased NO (p < 0.01) and AchE levels (p < 0.001) in the Acr-injured brains. Herniarin ameliorated Acr-induced brain injury via modulating redox hemostasis, cholinergic function, as well as inhibiting neuro-inflammation in rats.

Effect of green tea or black tea extract on lipid and protein oxidation in Cantonese sausage.

Natural polyphenols offer a safer alternative to synthetic antioxidants in meat products. This study investigated the efficacy of green tea and black tea extracts as natural antioxidants in Cantonese sausages to inhibit lipid and protein oxidation. Sausages were prepared with the addition of different concentrations - 100, 300, and 600 mg kg-1 total polyphenols (TP) - of green tea or black tea extract. Oxidation of the sausages was assessed through thiobarbituric acid reactants, carbonyl content, and thiol content, whereas consumer acceptability was evaluated based on texture, color, and sensory analysis. The tea extracts inhibited malondialdehyde production and reduced the thiobarbituric acid reactive substance value from 23.72 mmol MDA g-1 to less than 1.94 mmol MDA g-1. However, the addition of tea extracts decreased the thiol content and caused the loss of myosin heavy chain and actin bonds in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Although the addition of tea extracts increased the redness and hardness of the sausage, no significant difference in consumer acceptance between the control and treatment groups was observed in the sensory analysis. The tea extract inhibited the oxidation of lipids in Cantonese sausage. There was no negative effect on the sensory characteristics of sausages. The use of tea extracts as natural antioxidants in Cantonese sausage is therefore feasible. © 2024 Society of Chemical Industry.

8624 Maternal ZIKV Infection Causes Oxidative Damage in Placenta of Both Female and Male Mice

Abstract Disclosure: V.M. Nascimento: None. I.S. Andrade: None. C.B. Andrade: None. R.S. Alves: None. R.P. Sousa: None. A.C. Valim: None. L.S. Dias: None. K.N. Fontes: None. I.F. Miranda: None. S.A. Coelho: None. L.B. Arruda: None. R.S. Fortunato: None. T.M. Ortiga-Carvalho: None. The placenta is a maternal-fetal organ with nutritional, exchanging, hormonal and protecting functions that are vital for fetal growth and development during gestation. However, these functions can be disturbed by vertically transmitted pathogens such as Zika virus (ZIKV). In 2015, ZIKV was responsible for an increasing number of newborns with congenital abnormalities in Brazil, known as Congenital Zika Syndrome (CZS). Nonetheless, the extension of ZIKV infection in the placenta, leading to CZS, is still unclear. It is known that ZIKV causes oxidative stress in trophoblasts. However, no one has ever studied placental oxidative stress caused by ZIKV infection in vivo. Here, we investigated whether maternal ZIKV infection could cause sex-specific oxidative stress in murine placenta. At gestational day (GD)12.5, pregnant mice (C57BL/6; 8-12 weeks old) were infected with ZIKV-BRPE243 (5x107 PFU; ZIKV group, n=22) or Mock (supernatants of noninfected C6/36 cells; control group, n=28). At GD18.5, c-section was performed and maternal spleens, placentas, fetuses and fetal brains were collected. We measured placental carbonylated protein levels (n=7), by 2D OxyBlot, nitrotyrosine residues (n=5), by immunohistochemistry, reduced thiol content (n=9), total superoxide dismutase (SOD; n=9) and total glutathione peroxidase (GPx; n=9) activities, by spectrophotometry. Data were analyzed by unpaired Student’s t test and Two-Way ANOVA with Tukey’s post test and represented as mean±SEM. ZIKV infection increased maternal spleen weight (p&amp;lt;0.0001) and decreased both female (p=0.0429) and male (p=0.0171) placental weight. Although placental weights were affected by the infection, female and male fetal weight and fetal crown-rump length were not changed. Conversely, maternal ZIKV infection increased only female fetal brain weight (p=0.0028). Moreover, we found that ZIKV infection increased nitrotyrosine residues in the placental labyrinth region of both female (p&amp;lt;0.0001) and male (p=0.0007) fetuses. Meanwhile, nitrotyrosine residues were increased by the infection only in the placental junctional zone of female fetuses (p=0.0131). Although ZIKV increased this oxidative damage biomarker, placental carbonylated protein levels were not changed by the infection in both sexes. Also, reduced thiol content and total SOD and GPx activities were not affected. Together, our data show that maternal ZIKV infection caused oxidative stress in both placental regions of female and male fetuses, as we saw that ZIKV increased oxidative damage, although antioxidant defenses were not changed by the infection, at least at term (GD18.5). ZIKV infection may have disturbed placental growth, as we found that placental weight was decreased in both sexes. Moreover, we believe that the increase in the female fetal brain weight could be a suggestive feature of cerebral edema or hydrocephalus. Presentation: 6/2/2024

8624 Maternal ZIKV Infection Causes Oxidative Damage in Placenta of Both Female and Male Mice

Abstract Disclosure: V.M. Nascimento: None. I.S. Andrade: None. C.B. Andrade: None. R.S. Alves: None. R.P. Sousa: None. A.C. Valim: None. L.S. Dias: None. K.N. Fontes: None. I.F. Miranda: None. S.A. Coelho: None. L.B. Arruda: None. R.S. Fortunato: None. T.M. Ortiga-Carvalho: None. The placenta is a maternal-fetal organ with nutritional, exchanging, hormonal and protecting functions that are vital for fetal growth and development during gestation. However, these functions can be disturbed by vertically transmitted pathogens such as Zika virus (ZIKV). In 2015, ZIKV was responsible for an increasing number of newborns with congenital abnormalities in Brazil, known as Congenital Zika Syndrome (CZS). Nonetheless, the extension of ZIKV infection in the placenta, leading to CZS, is still unclear. It is known that ZIKV causes oxidative stress in trophoblasts. However, no one has ever studied placental oxidative stress caused by ZIKV infection in vivo. Here, we investigated whether maternal ZIKV infection could cause sex-specific oxidative stress in murine placenta. At gestational day (GD)12.5, pregnant mice (C57BL/6; 8-12 weeks old) were infected with ZIKV-BRPE243 (5x107 PFU; ZIKV group, n=22) or Mock (supernatants of noninfected C6/36 cells; control group, n=28). At GD18.5, c-section was performed and maternal spleens, placentas, fetuses and fetal brains were collected. We measured placental carbonylated protein levels (n=7), by 2D OxyBlot, nitrotyrosine residues (n=5), by immunohistochemistry, reduced thiol content (n=9), total superoxide dismutase (SOD; n=9) and total glutathione peroxidase (GPx; n=9) activities, by spectrophotometry. Data were analyzed by unpaired Student’s t test and Two-Way ANOVA with Tukey’s post test and represented as mean±SEM. ZIKV infection increased maternal spleen weight (p&amp;lt;0.0001) and decreased both female (p=0.0429) and male (p=0.0171) placental weight. Although placental weights were affected by the infection, female and male fetal weight and fetal crown-rump length were not changed. Conversely, maternal ZIKV infection increased only female fetal brain weight (p=0.0028). Moreover, we found that ZIKV infection increased nitrotyrosine residues in the placental labyrinth region of both female (p&amp;lt;0.0001) and male (p=0.0007) fetuses. Meanwhile, nitrotyrosine residues were increased by the infection only in the placental junctional zone of female fetuses (p=0.0131). Although ZIKV increased this oxidative damage biomarker, placental carbonylated protein levels were not changed by the infection in both sexes. Also, reduced thiol content and total SOD and GPx activities were not affected. Together, our data show that maternal ZIKV infection caused oxidative stress in both placental regions of female and male fetuses, as we saw that ZIKV increased oxidative damage, although antioxidant defenses were not changed by the infection, at least at term (GD18.5). ZIKV infection may have disturbed placental growth, as we found that placental weight was decreased in both sexes. Moreover, we believe that the increase in the female fetal brain weight could be a suggestive feature of cerebral edema or hydrocephalus. Presentation: 6/2/2024

The Antidiabetic Drug Metformin Attenuated Depressive and Anxietylike Behaviors and Oxidative Stress in the Brain in a Rodent Model of Inflammation Induced by Lipopolysaccharide in Male Rats.

Inflammation is considered to be a link between diabetes and central nervous system (CNS) disorders, including depression and anxiety. Metformin is suggested to have antioxidant, anti-inflammatory, and mood-improving effects. The aim of the current research was to investigate the effects of the antidiabetic drug metformin on depressive- and anxiety- like behaviors and oxidative stress in the brain in a rodent model of inflammation induced by lipopolysaccharide (LPS) in male rats. The rats were treated as follows: (1) Vehicle instead of metformin and lipopolysaccharide, (2) Lipopolysaccharide (1 mg/ kg) + vehicle instead of metformin, (3-5) Lipopolysaccharide + 50, 100, or 150 mg/ kg of metformin. After the behavioral tests, including open field (OF), elevated pulse maze (EPM), and force swimming (FS) tests, the brains were removed, and malondialdehyde (MDA), nitric oxide (NO) metabolites, total thiol, catalase (CAT) activity, interleukin-6 (IL-6) and superoxide dismutase (SOD) activity were determined. In the EPM, metformin increased the open arm time and entry and decreased closed arm time and entry. In the FS test, metformin lowered the immobility and increased active time compared to lipopolysaccharide. In the OF test, metformin increased total crossing and total distance, time spent, traveled distance, and crossing number in the central zone. As a result of metformin administration, IL-6, MDA, and NO metabolites were decreased while thiol content, SOD, and CAT activity were increased. The results indicated that the well-known antidiabetic drug metformin attenuated depressive- and anxiety-like behaviors induced by inflammation in rats. These beneficial effects are suggested to be due to their attenuating effects on neuroinflammation, oxidative stress, and NO in the brain.

Multiple Biomarker Responses in Aegla castro Exposed to Copper: A Laboratory Approach.

Although some biomarkers have already been determined in aeglids collected in the field, data from laboratory exposures are scarce. To our knowledge, no studies have investigated oxidative stress biomarkers in aeglids exposed to metals in the laboratory, or performed hemocyte counts and the comet assay using gill and hepatopancreas of aeglids. Thus, we investigated the effects of acute Cu exposure on intermolt males of Aegla castro, collected from a reference stream, acclimated for 6days in the laboratory, and then exposed to 11μgL-1 of dissolved Cu (Cu 11) or only to water (CTR), for 24h. Gill and hepatopancreas samples were used to determine Cu accumulation, DNA damage, and metallothionein content (MT), while hemolymph samples were used to determine Cu accumulation, DNA damage, and hemocyte counts. Muscle samples were used to determine Cu accumulation and acetylcholinesterase activity (AChE). Non-protein thiol content (NPSH), catalase (CAT), glutathione S-transferase activities (GST), lipoperoxidation (LPO), and protein carbonylation content (PCC) were measured only in the hepatopancreas. Aegla castro exposed to Cu accumulated this metal in gills and activated detoxification mechanisms, through increased MT content in the gill, and showed an immune response, evidenced by an increase in hyaline hemocytes. Therefore, gill and hemocytes appear to have a protective role in preventing the transport and bioavailability of Cu through the body. On the other hand, we observed a decrease in MT content in the hepatopancreas of crabs exposed to Cu, suggesting the excretion of MT in association with Cu bound to the sulfhydryl groups of this protein.