Markers Of Antioxidant Activity Research Articles

Revealing quality chemicals of Tetrastigma hemsleyanum roots in different geographical origins using untargeted metabolomics and random-forest based spectrum-effect analysis

Tetrastigma hemsleyanum root is a popular functional food in China, and the price varies based on the origin of the product. The link between the origin, metabolic profile, and bioactivity of T. hemsleyanum must be investigated. This study compares the metabolic profiles of 254 samples collected from eight different areas with 49 potential key chemical markers using plant metabolomics. The metabolic pathways of the five critical flavonoid metabolites were annotated and enriched using the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. Moreover, a random forest model aiding the spectrum-effect relationship analysis was developed for the first time indicating catechin and darendoside B as potential quality markers of antioxidant activity. The findings of this study provide a comprehensive understanding of the chemical composition and bioactive compounds of T. hemsleyanum as well as valuable information on the evaluation of the quality of various samples and products in the market.

Research on the quality markers of antioxidant activity of Kai-Xin-San based on the spectrum-effect relationship.

Background: Kai-Xin-San (KXS) is one of the classic famous traditional Chinese medicine prescriptions for amnesia, which has been applied for thousands of years. Modern pharmacological research has found that KXS has significant therapeutic efficacy on nervous system diseases, which is related to its antioxidant activity. However, the antioxidant material basis and quality markers (Q-makers) of KXS have not been studied. Objective: The objective of this study is to explore the Q-makers of antioxidant activity of KXS based on spectrum-effect relationship. Methods: Specifically, the metabolites in KXS extracts were identified by UPLC-Q-Exactive Orbitrap MS/MS. The fingerprint profile of KXS extracts were established by high-performance liquid chromatography (HPLC) and seven common peaks were identified. Meanwhile, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) test was used to evaluate the free radical scavenging ability of KXS. The spectrum-effect relationship between its HPLC fingerprint and DPPH free radical scavenging activity was preliminarily examined by the Pearson correlation analysis, grey relation analysis (GRA), and orthogonal partial least squares discrimination analysis (OPLS-DA). Further, the antioxidant effect of KXS and its Q-makers were validated through human neuroblastoma (SH-SY5Y) cells experiment. Results: The results showed that 103 metabolites were identified from KXS, and the similarity values between HPLC fingerprint of twelve batches of KXS were greater than 0.900. At the same time, the results of Pearson correlation analysis showed that the peaks 8, 1, 14, 17, 18, 24, 16, 21, 15, 13, 6, 5, and 3 from KXS were positively correlated with the scavenging activity values of DPPH. Combined with the results of GRA and OPLS-DA, peaks 1, 3, 5 (Sibiricose A6), 6, 13 (Ginsenoside Rg1), 15, and 24 in the fingerprints were screen out as the potential Q-makers of KXS for antioxidant effect. Besides, the results of CCK-8 assay showed that KXS and its Q-makers remarkably reduced the oxidative damage of SH-SY5Y cells caused by H2O2. However, the antioxidant activity of KXS was decreased significantly after Q-makers were knocked out. Conclusion: In conclusion, the metabolites in KXS were successfully identified by UPLC-Q-Exactive Orbitrap MS/MS, and the Q-makers of KXS for antioxidant effect was analyzed based on the spectrum-effect relationship. These results are beneficial to clarify the antioxidant material basis of KXS and provide the quality control standards for new KXS products development.

Validation of an alternative quantitative method for determination of antioxidant potential in Rosmarinus officinalis L. and Chilli pepper

AbstractWe provide proof-of-concept for the innovative method of 2,6-dichloroindophenol (DCPIP) for measuring the antioxidant activity of plant extracts. Antioxidant content can be determined using the standard DCPIP test and compare the results with the DPPH results as conventional method. DCPIP operates on the premise that the deep blue color of the oxidized dye is reduced to an invisible solution. Ascorbic acid’s antioxidant activity was measured by DCPIP at different times (from 1 to 60 min) and was concentration-dependent, with the maximum activity being at 400 g/mL. In addition, when compared to other incubation durations, the ascorbic acid standard, a natural antioxidant, gave the maximum activity within the first five minutes of incubation with DCPIP. DCPIP is a marker of antioxidant activity both against vitamin C and plant extracts. The DCPIP approach is quick and unaffected by pH variation. The stability of the DCPIP reagent over time (5 and 30 min) and color reduction by ascorbic acid as a natural antioxidant standard were demonstrated using a straightforward and quick method.

Effect of Copper and Zinc Ions on Biochemical and Molecular Characteristics of Calcium Oxalate Renal Stones: a Controlled Clinical Study.

Contradictory results are existed in the literature regarding the impact of trace elements on the pathogenesis of calcium oxalate (CaOx) stone patients. Therefore, the aim of our study was to investigate the effect of Cu and Zn on biochemical and molecular characteristics of CaOx stones. Plasma and urine concentrations of Cu and Zn in 30 CaOx stones patients and 20 controls were determined by flame atomic absorption spectrometry (FAAS). Urinary levels of citric acid and oxalate were measured by commercial spectrophotometric kits. Blood levels of glutathione reduced (GSH) and catalase (CAT) were determined as markers of antioxidant activity, while blood malondialdehyde (MDA) and urine level of nitric oxide (NO) were used to assess oxidative stress. Gene expression of MAPk pathway (ERK, P38, and JNK) were estimated. The plasma and urine levels of Cu were significantly increased in the patient group compared to those of controls, while the levels of Zn were decreased. Excessive urinary excretion of citric acid and oxalate were found among CaOx stone patients. The GSH and CAT concentration were significantly reduced in CaOx stones patients compared to healthy group. The plasma MDA and urine NO concentration were significantly increased in CaOx stones patients compared to control group. The expressions of the studied genes were significantly increased in CaOx stones patients. These findings suggest that alteration in Cu and Zn might contribute to pathogenesis of CaOx patients through oxidative stress and MAPK pathway genes (ERK, P38 and JNK).

Lycopene ameliorates DEHP exposure-induced renal pyroptosis through the Nrf2/Keap-1/NLRP3/Caspase-1 axis

Di (2-ethylhexyl) phthalate (DEHP) is commonly used as a plasticizer in plastic products, and due to its unique chemical composition, it frequently dissolves and enters the environment. Lycopene as a natural carotenoid has been shown to have powerful antioxidant capacity and strong kidney protection. This study aimed to investigate the role of the interplay between oxidative stress and the classical pyroptosis pathway in LYC alleviating DEHP-induced renal injury. ICR mice were given DEHP (500 mg/kg/d or 1000 mg/kg/d) and/or LYC (5 mg/kg/d) for 28 days to explore the underlying mechanisms of this hypothesis. Our results indicated that DEHP caused the shedding of renal tubular epithelial cells, increased the content of kidney injury molecule-1 (Kim-1) and neutrophil gelatinase-associated lipocalin (NGAL) in the tissue, the decrease of antioxidant activity markers and the increase of oxidative stress indexes. It is gratifying that LYC alleviates DEHP-induced renal injury. The expression of nuclear factor erythrocyte 2-related factor 2 (Nrf2) and its downstream target genes is improved in DEHP induced renal injury through LYC mediated protection. Meanwhile, LYC supplementation can inhibit DEHP-induced Caspase-1/NLRP3-dependent pyroptosis and inflammatory responses. Taken together, DEHP administration resulted in nephrotoxicity, but these changes ameliorated by LYC may through crosstalk between the Nrf2/Keap-1/NLRP3/Caspase-1 pathway. Our study provides new evidence that LYC protects against kidney injury caused by DEHP.

Protective role of curcumin on aflatoxin B1-induced TLR4/RIPK pathway mediated-necroptosis and inflammation in chicken liver

This study set out to assess the mitigative effects of curcumin on AFB1-induced necroptosis and inflammation in chicken liver. Ninety-six one-day-old AA broiler chickens were separated into four groups, including control group, AFB1 (1 mg/kg) group, curcumin (300 mg/kg) + AFB1 (1 mg/kg) group and curcumin (300 mg/kg) group. After 28 days treatment, livers were collected for different experimental analyses. The morphological observation results showed obvious necrotic characteristics, including cell swelling, rupture of cell and mitochondrial membranes and inflammation in chicken livers. AFB1 exposure increased oxidative stress index (ROS and MDA) and decreased the antioxidant activity markers (SOD, CAT and GSH) and ATPase activities in chickens’ liver. ELISA results showed that AFB1 exposure significantly induced the cytokines (TNF-α, iNOS, IL-6 and IL-1β) release from the liver tissues. While, western blot and qRT-PCR results showed that the protein and mRNA expressions of inflammatory (TLR4/myd88/NF-κB) and necroptosis (RIPK1/RIPK3/MLKL) genes were up-regulated by AFB1 exposure. We suspect that signal crosstalk between TLR4 and TNF-α triggers inflammation and RIPK1/RIPK3 mediating necroptosis in AFB1-induced chicken liver injury. Curcumin can regulate the TLR4/RIPK signaling pathway, reduced oxidative stress biomarkers and inflammatory cytokines levels and attenuated the expression of necroptosis and inflammation genes altered by AFB1 to reduce necroptosis of chicken liver tissue. In conclusion, curcumin can protect against AFB1-induced necroptosis and inflammation by TLR4/RIPK pathway in chicken liver.

Nanoencapsulated Extract of a Red Seaweed (Rhodophyta) Species as a Promising Source of Natural Antioxidants.

Marine seaweed species represent an important source of bioactive compounds possessing antioxidant activity. This study aimed at evaluating the antioxidant capacity of the Jania rubens algal extract by means of two antioxidant assays, i.e., 2,2-diphenyl-1-picrylhydrazyl and ferric-reducing antioxidant power. The seaweeds’ total phenolic and flavonoid contents were also assayed as markers of antioxidant activity. To identify active agents responsible for the antioxidant activity, gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry were used for comprehensive metabolites characterization. To enhance the Jania rubens efficacy, the extract was nanoencapsulated using an ionic gelation method by means of high-pressure homogenization. The optimum nanoformulation had a particle size of 161 nm, a ζ potential of 31.2 mV, a polydispersity index of 0.211, and entrapment efficiency of 99.7%. The in vitro phytochemicals’ release profiles of Jania rubens chitosan nanoparticles in comparison to the concentration of the raw algal extract were studied by the dialysis bag diffusion method revealing that the extract was released in a controlled pattern. The results indicated the potential advantages of the encapsulated Jania rubens extract, with its potent antioxidant activity, for use in different applications where sustained release is useful.

Association between markers of oxidative stress and cognitive functioning in schizophrenia

High levels of oxidative stress have been implicated in the pathogenesis of schizophrenia. Our objectives were to review the association between oxidative stress markers and neurocognitive functioning in schizophrenia. Electronic search of MEDLINE, ProQuest, Scopus, and Google Scholar databases was carried out in September 2022 to identify relevant English language articles. Articles identified were grouped under the following themes: comparison of oxidative stress or antioxidant biomarkers between patients with schizophrenia and general population, association between biomarkers of oxidative stress and cognitive dysfunction in schizophrenia, and clinical trials evaluating the effect of antioxidant compounds on cognitive functioning in schizophrenia. We reviewed 36 eligible papers: 30 observational studies and 6 intervention trials. Marked heterogeneity was noted in patient population, oxidative stress markers examined, and cognitive measures. Majority (83.3%) of the observational studies were cross-sectional in design; half of them (50%) were conducted on chronic, stable, medicated patients. Most of the observational studies showed increased levels of oxidative stress biomarkers and decreased antioxidant activity in schizophrenia compared to controls. A significant positive association was noted between markers of antioxidant activity and cognitive performance in this group. Treatment trials of antioxidant agents showed positive but inconsistent evidence for benefits on cognitive measures and clinical symptomatology in schizophrenia. Our findings are broadly consistent with the redox imbalance hypothesis in schizophrenia. The significant associations noted between antioxidant markers and cognitive functioning may have key neurobiological and therapeutic implications.

The effect of complex herbal extract and methotrexate on suppressing adjuvant arthritis in rats

The present study evaluated the therapeutic benefits of complex herbal preparation named CBMDS, consisting of turmeric (Curcuma longa), Boswellia (Boswellia serrata), Methylsulphonylmethane, Devil’s Claw (Harpagophytum procumbens) and Silymarin, using it in combination with methotrexate, in order to suppress adjuvant arthritis in rats, and to attenuate methotrexate-induced liver damage. Adjuvant arthritis was induced in 28 rats by a single subplantar injection of complete Freund’s adjuvant (0.1 mL) into the left hind paw. The animals were divided into four groups (with seven animals in each). Group I received CBMDS, Group II - CBMDS in combination with methotrexate, and Group III just methotrexate. The treatment lasted from day 0 to day 17 (CBMDS was given daily except weekends in a dose of 160 mg/kg, methotrexate - 2 mg/kg once a week). Group IV was the control group. Clinical (body weight, hind paw volume, erythrocyte sedimentation rate, leukocyte count), biochemical (serum pro-/antioxidant activity markers), immunological (serum interleukin levels) and histological changes in joint and liver tissues were evaluated. CBMDS significantly alleviated arthritis and reduced hepatic damage, which was more evident in the methotrexate group. The combined treatment also markedly reduced arthritic symptoms and levels of malondialdehyde. Antioxidant activity was significantly higher in treated Groups I and II. CBMDS and its combination with methotrexate promoted anti-arthritic action, reduced histological changes in the joint tissues, and minimized methotrexate-induced liver toxicity.

Nonaqueous Polyethylene Glycol as a Safer Alternative to Ethanolic Propolis Extracts with Comparable Antioxidant and Antimicrobial Activity.

We compared the chemical composition, antioxidant and antimicrobial activity of two propolis extracts: one obtained with nonaqueous polyethylene glycol, PEG 400 (PgEP), and the other obtained with ethanol (EEP). We analyzed the total phenolic content (TPC) and the concentrations of ten markers of propolis antioxidant activity with HPLC-UV: caffeic acid, p-coumaric acid, trans-ferulic acid, trans-cinnamic acid, kaempferol, apigenin, pinocembrin, chrysin, CAPE, and galangin. Antioxidant activity was tested using DPPH and FRAP assay, and antimicrobial activity was assessed through minimum inhibitory concentrations (MICs) and minimum biofilm eradication concentration (MBEC) determination. Maceration gave the yield of propolis of 25.2 ± 0.08% in EEP, and 21.5 ± 0.24% in PgEP. All ten markers of antioxidant activity were found in both extracts, with all marker concentrations, except kaempferol, higher in EEP. There was no significant difference between the TPC and antioxidant activity of the PgEP and the EEP extract; TPC of PgEP was 16.78 ± 0.23 mg/mL, while EEP had TPC of 15.92 ± 0.78 mg/mL. Both extracts had antimicrobial activity against most investigated pathogens and Staphylococcus aureus, Acinetobacter baumannii, and Escherichia coli biofilms. EEP was more effective against all tested susceptible pathogens, except E. coli, possibly due to higher content of kaempferol in PgEP relative to other polyphenols. Nonaqueous PEG 400 could be used for propolis extraction. It gives extracts with comparable concentrations of antioxidants and has a good antioxidant and antimicrobial activity. It is a safe excipient, convenient for pediatric and veterinary formulations.

Lycopene prevents DEHP-induced hepatic oxidative stress damage by crosstalk between AHR–Nrf2 pathway

Di (2-ethylhexyl) phthalate (DEHP) is a widespread plasticizer that persists in the environment and can significantly contribute to serious health hazards of liver especially oxidative stress injury. Lycopene (LYC) as a carotenoid has recently gained widespread attention because of antioxidant activity. However, the potential mechanism of DEHP-induced hepatotoxicity and antagonism effect of LYC on it are still unclear. To explore the underlying mechanisms of this hypothesis, the mice were given by gavage with LYC (5 mg/kg) and DEHP (500 or 1000 mg/kg). The data suggested that DEHP caused liver enlargement, reduction of antioxidant activity markers, increase of oxidative stress indicators and disorder of cytochrome P450 enzymes system (CYP450s) homeostasis. DEHP-induced reactive oxygen species (ROS) activated the NF-E2-relatedfactor2 (Nrf2) and nuclear xenobiotic receptors (NXRs) system including Aryl hydrocarbon receptor (AHR), Pregnane X receptor (PXR) and Constitutive androstane receptor (CAR). Interestingly, these disorders and injuries were prevented after LYC treatment. Taken together, DEHP administration resulted in hepatotoxicity including oxidative stress injury and disordered CYP450 system, but these alterations might be ameliorated by LYC via crosstalk between AHR–Nrf2 pathway.

Phenolic Contents and Antioxidant Activity of Helicteres Hirsuta Extracts

Helicteres hirsuta, which belongs to the Malvaceae family, is used in traditional medicine to treat malaria, diabetes and cervical cancer. While these uses have not been validated in clinical studies thus far, extracts from H. hirsuta exhibit confirmed antioxidant and potential anti-cancer activity that warrant critical assessment. The aim of this study was to determine the antioxidant properties of different parts of the plant H. hirsuta. The antioxidant capacities of methanol extracts of different plant parts, and specific solvent fractions thereof were determined on the basis of 2,2-diphenyl−1−picrylhydrazyl (DPPH) radical scavenging and total antioxidant activity assays. The ethyl acetate fraction from the leaf material of H. hirsuta has the most potent antioxidant activity with the lowest half maximal inhibitory concentration (IC50) value of 9.50 μg/mL, that is 4 times lower than the IC50 of curcumin. The amounts of methyl gallate and rutin dominated the extracts at 8.62 ± 0.01 and 6.76 ± 0.02 mg/g, respectively, out of a total of 21.00 ± 0.01 mg/g active phenolic fraction. The amount of methyl gallate strongly correlated with total phenolic content and total antioxidant capacity, thus this compound may be useful as a marker of antioxidant activity in common medicinal plants.

Phenolic Fractions from Vaccinium vitis-idaea L. and Their Antioxidant and Anticancer Activities Assessment.

Lingonberry leaves and fruits are associated with a range of potential bioactivities related to their phenolic content and composition, but the identification of major biological activity markers remains limited. The present study aimed at the isolation of lingonberry phenolic fractions and biological activity evaluation of them. Crude dry extracts of lingonberry leaves and fruits were fractionated by chromatography using Sephadex LH-20 and analyzed by validated HPLC-PDA method. For each fraction, the anticancer activity against human clear cell renal cell carcinoma (CaKi-1), human colon adenocarcinoma (HT-29), and human malignant melanoma (IGR39) cell lines was determined using MTT assay, and the radical scavenging, reducing, and chelating activities were investigated using ABTS, FRAP, and FIC assays, respectively. Further, 28 phenolics were identified and quantified in the crude extract of lingonberry leaves and 37 in the extract of fruits. These compounds, during fractionation steps, were selectively eluted into active fractions, enriched with different groups of phenolics—monophenols, anthocyanins, phenolic acids, catechins, flavonols, or proanthocyanidins. Fractions of lingonberry leaves and fruits, obtained by the last fractionation step, proved to be the most active against tested cancer cell lines and possessed the greatest antioxidant activity. In this perspective, the predominant compounds of these fractions—polymeric and mainly A-type dimeric proanthocyanidins—also quercetin can be considered to be anticancer and antioxidant activity markers of lingonberries.

Análise do estresse oxidativo em ratos Wistar submetidos a treina-mento intervalado de alta intensidade

Physical exercise is a known factor that can promote oxidative stress and may result in cellular damage if not neutralized by antioxidant mechanisms. The aim of this study was to determine if the level of hepatic oxidative stress resulting from high-intensity interval training (HIIT) is affected by the frequency pattern (consecutive vs. non-consecutive) of the training sessions. Thirty-two Wistar rats were divided equally into four groups: two control groups, (CS1) and (CS2), remained sedentary throughout the experiment, and two test groups, (CT1) and (CT2), and were submitted to HIIT for 12 consecutive and nonconsecutive (12 sessions, 3 times/wk over four weeks) days, respectively. There were no significant differences in markers of oxidative damage measured in hepatic tissue (TBARS) and markers of antioxidant activity (Sulfhydryl, FRAP), as well as markers of hepatic damage (AST and ALT) and antioxidant defense (Uric Acid) measured in plasma of both HIIT groups after training compared to the control groups. The results indicate that both HIIT performed for 12 consecutive and nonconsecutive sessions did not promote hepatic oxidative damage in rats.

Consumption of Anthocyanin-Rich Mulberry Fruit Jelly with a High-Fat Meal Decreases Postprandial Serum Cardiometabolic Risk Factors in Dyslipidemia Subjects

Anthocyanin content in berries has been reported to promote antioxidant properties that mitigate the occurrence of noncommunicable diseases. However, only a few studies have investigated the benefits of anthocyanin-rich food products from mulberry fruit to reduce the cardiometabolic risk factor in dyslipidemia subjects. Anthocyanin-rich mulberry fruit jelly was formulated using mulberry fruit powder (MFP), and its activities on serum cardiometabolic risk factors in dyslipidemia subjects were studied. Morus alba var. Chiang Mai was used as the ingredient for MFP jelly containing 14 g MFP (191 mg anthocyanin) per serving size (170 g). To investigate the effect of MFP jelly on reduction of cardiometabolic risk factors, sixteen dyslipidemia subjects were given one serving of MFP jelly every day for seven days. After MFP jelly intervention, fasting blood cholesterol, low-density lipoprotein (LDL), and inflammatory markers including interleukin-6 levels of the subjects were significantly lower. Postprandial blood parameters were measured at 0–240 min after consuming a high-fat meal before and after MFP jelly intervention. Postprandial blood glucose at 30 min (p<0.05) and insulin at 60 and 90 min (p<0.01) were lower in MFP than in placebo jelly. The area under the curve of insulin in MFP jelly was smaller than in placebo by 31.2%. Therefore, MFP jelly intervention increased insulin sensitivity. For antioxidant activity markers, postprandial oxygen radical absorbance capacity after MFP jelly intervention gave a smaller decrease after high-fat meal intake compared to after placebo jelly intervention. Moreover, for the oxidative stress markers, postprandial malondialdehyde level was significantly lower in MFP jelly. Seven days of intervention by one serving size of MFP jelly containing 191 mg of anthocyanins reduced cardiometabolic risk factors by lowering blood total cholesterol, LDL, and inflammation, and improving insulin sensitivity and postprandial blood antioxidant-oxidative stress activity in dyslipidemia subjects. This trial is registered with TCTR20200415003.

A novel antifibrotic strategy utilizing conditioned media obtained from miR-150-transfected adipose-derived stem cells: validation of an animal model of liver fibrosis

The limitations of stem cells have led researchers to investigate the secretome, which is the secretory materials in stem cells, since the principal mechanism of action of stem cells is mediated by the secretome. In this study, we determined the antifibrotic potential of the secretome released from miR-150-transfected adipose-derived stromal cells (ASCs). The secretome released from ASCs that were transfected with antifibrotic miR-150 was obtained (referred to as the miR-150 secretome). To validate the antifibrotic effects of the miR-150 secretome, we generated in vitro and in vivo models of liver fibrosis by treating human hepatic stellate cells (LX2 cells) with thioacetamide (TAA) and subcutaneous injection of TAA into mice, respectively. In the in vitro model, more significant reductions in the expression of fibrosis-related markers, such as TGFβ, Col1A1, and α-SMA, were observed by using the miR-150 secretome than the control secretome, specifically in TAA-treated LX2 cells. In the in vivo model, infusion of the miR-150 secretome into mice with liver fibrosis abrogated the increase in serum levels of systemic inflammatory cytokines, such as IL-6 and TNF-α, and induced increased expression of antifibrotic, proliferation, and antioxidant activity markers in the liver. Our in vitro and in vivo experiments indicate that the miR-150 secretome is superior to the naive secretome in terms of ameliorating liver fibrosis, minimizing systemic inflammatory responses, and promoting antioxidant enzyme expression. Therefore, we conclude that miR-150 transfection into ASCs has the potential to induce the release of secretory materials with enhanced antifibrotic, proliferative, and antioxidant properties.

Astilbin protects chicken peripheral blood lymphocytes from cadmium-induced necroptosis via oxidative stress and the PI3K/Akt pathway.

Astilbin (ASB), a dihydroflavonol glycoside, is widely found in a variety of plants and in functional foods and acts as a powerful antioxidant. The aim of this study was to investigate the underlying mechanisms involved in the antagonistic effects of ASB on cadmium (Cd)-induced necroptosis in chicken peripheral blood lymphocytes. Peripheral blood lymphocytes were aseptically collected from Roman white hens and then randomly divided into five groups: the control group was incubated without additional reagents, while the other groups were incubated with Cd, ASB, a combination of Cd and ASB, and 0.1% DMSO. After a 24h treatment, cell samples were collected. The results showed that some morphological changes consistent with necroptosis were observed in the Cd-treated groups, suggesting the occurrence of necroptosis. Simultaneously, antioxidant activity markers (CAT, SOD, GSH, GSH-px, and T-AOC) decreased and indicators of oxidative stress (MDA, iNOS, NO, H2O2, ·OH and ROS) increased. The production of ROS induced the activation of the PI3K/Akt signaling pathway, as the expression levels of PI3K, Akt and PDK1 were significantly elevated. Additionally, the expression levels of RIPK3, RIPK1, MLKL, TAK1, TAB2 and TAB3 were increased and that of Caspase-8 was decreased, which could cause the necroptosis. However, the most important our results was that ASB supplements remarkably attenuated the Cd-induced effects. We conclude that the Cd treatment promoted an imbalance of the antioxidant status and activated the PI3K/Akt pathway, leading to necroptosis in chicken peripheral blood lymphocytes, and that ASB was able to partially ameliorate the effect of Cd-induced necroptosis.

Efficacy of sodium butyrate in alleviating mammary oxidative stress induced by sub-acute ruminal acidosis in lactating goats

Sub-acute ruminal acidosis (SARA) [1] is one of the most common problems of dairy animals causing great economical loss due to decreased milk production. Here we determined the antioxidant effect of sodium butyrate (NaB) [2] in experimentally induced SARA and its effects on mammary epithelial tissues of goat. Goats (n = 12) were equally divided into two groups: high-concentrate (HC) as control group fed with HC diet (concentrate: forage = 6:4) whereas HC + NaB as treatment group fed HC diet with NaB at 1% by weight for 24 weeks. Mammary epithelial tissue samples were analyzed for the expression of genes and proteins responsible for oxidative stress as well as biochemical markers of antioxidant activity in the form of Reactive Oxygen Species (ROS). The total antioxidant capacity (T-AOC) of antioxidant enzymes was also calculated. Butyrate induced antioxidant effect by increasing mRNA and protein abundance of antioxidants in mammary gland of HC + NaB group compared to HC group. Likewise, the total antioxidant capacity (T-AOC) was significantly increased and Malondialdehyde (MDA) concentration was decreased in HC + NaB group compared to HC group. It is concluded that oxidative stress in mammary gland of goats induced by high concentrate diet was alleviated by NaB supplementation.

Different Extraction Solvents and Sample Preparation Methods Affect the Antioxidant Activities and Total Phenol Contents of Theobroma Cacao Powder (P01-016-19)

ObjectivesCacao(Theobroma cacao) has been widely consumed since more than 4000 years ago which was regarded as sacred food. Cacao has been considered as a strong antioxidant due to its abundant phenolic and especially flavonoid contents. In this experiment, different extraction solvents and methods were used to analyze whether these factors affect the antioxidant activities and phytochemical compounds of cacao powder extracts. MethodsFour different cacao extract samples were examined to identify their anti-oxidative activities; raw cacao powder extracted in 70% methanol solution(RM), raw cacao powder extracted in 80% ethanol solution(RE), freeze dried cacao powder extracted in 70% methanol solution(FM), freeze dried cacao powder extracted in 80% ethanol solution(FE). Total phenolic compound content was calculated based on gallic acid standard curve and total flavonoid content was calculated based on the standard curve of quercetin. ABTS scavenging activity was experimented with ABTS reagent diluted by PBS(pH7.4) to make the O.D. value 0.7 ± 0.02 at 734 nm before using 2.5 mM ABTS. The DPPH radical scavenging activity was assessed by 1 mM DPPH methanol solution. ResultsThe contents of total phenol compounds were 45.4 mg GAE/g(RM), 37.3 mg GAE/g(RE), 50.6 mg GAE/g(FM), and 52.7 mg GAE/g(FE). The contents of total flavonoid compounds were 34.3 mg QE/g(RM), 29.7 mg QE/g(RE), 85.1 mg QE/g(FM), and 89.7 mg QE/g(FE). FM group showed the highest ABTS radical scavenging activity which is a marker of antioxidant activity but the other sample groups represented almost as high ABTS radical scavenging activity as FM group. RM group showed the highest DPPH radical scavenging activity which is also a marker of antioxidant activity. ConclusionsThe results show that different extract solvents and sample preparation methods affect DPPH radical scavenging activity, total phenol and flavonoid contents of cacao powder extracts. Therefore, appropriate methods should be applied according to the purpose of the study. Funding SourcesDepartment of Food and Nutrition, Sookmyung Women’s University, Republic of Korea.

Hercynine content in widely consumed commercial beverages

Hercynine, the main biosynthetic precursor and oxidative metabolite of ergothioneine, was measured by an LC-ESI-MS/MS method in tea, coffee, beer, and wine samples. Results showed that hercynine was detectable and measurable in all beverage. Among teas, the higher concentration was in the black variety (170.45 ± 7.84 ng/mg of dry weight) followed by the white (130.63 ± 8.79 ng/mg of dry weight) and green ones (71.62 ± 6.13 and 47.43 ± 7.66 ng/mg of dry weight). Compared to teas, coffees had less amount of hercynine with more homogeneous levels that appear unlinked to the kind of mixture (100% Arabica 12.92 ± 1.01 ng/mg of dry weight, 50/50 Arabica/Robusta 10.52 ± 0.38 ng/mg of dry weight, and decaffeinated 8.59 ± 0.75 ng/mg of dry weight). Overall, the red wines had the highest values of hercynine compared to the whites (379.57 ± 238.15 vs. 575.51 ± 62.60 nmol/L). In the latter, hercynine concentrations showed a trend toward an increase with increasing pH values. Hercynine concentrations in the beers were similar to the levels in wines, and there was no difference between traditional and gluten-free sample (679.24 ± 0.92 vs. 570.58 ± 0.88 nmol/L). These data highlight the potential of hercynine as a possible marker of antioxidant activity of ergothioneine in beverages and food.