Free Radical Scavengers Superoxide Research Articles

Structural characteristics and antioxidant activities of a novel polysaccharide from Euphorbia himalayensis root

Euphorbia himalayensis Boiss. is an alpine member of the Euphorbiaceae family. Its dried roots have been used to treat digestive problems and chest congestion in traditional Tibetan and Mongolian medicine. Despite thousands of years of use in medicine, the bioactive compounds of the root remain unknown. Herein, we isolated a novel aqueous-soluble polysaccharide (EHP2) from the E. himalayensis root and determined its structural characteristics via high-performance gel permeation chromatography, Fourier-transform infrared spectroscopy, gas chromatography–mass spectrometry, and nuclear magnetic resonance spectrometry. The homogeneous molecular weight of EHP2 was 23.6 kDa with narrow polydisperity (Mw/Mn = 1.4), and EHP2 mainly comprised of glucose (86.4%), galactose (11.9%) and mannose (1.7%). The major backbone of EHP2 was →4)-α-D-GalAp-(1 → 4)-α-D-Glcp-(1 → and the branch chain was α-D-Glcp-(1→. The antioxidant activity of the EHP2 was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide anion radical scavenging assays, and antioxidant enzyme activity (SOD, GSH and MDA) was determined in human umbilical vein endothelial cells (HUVECs). The EHP2 demonstrated lower potential scavenging effects on DPPH and superoxide free radical scavenger than ascorbic acid, and in HUVECs, it led to increased SOD and GSH activities and decreased MDA levels. This study is the first to describe an E. himalayensis polysaccharide compound with potential antioxidant activity.

Comprehensive In Silico Analysis of Uncaria Tomentosa Extract: Chemical Profiling, Antioxidant Assessment, and CLASP Protein Interaction for Drug Design in Neurodegenerative Diseases.

Uncaria tomentosa is a traditional medicinal herb renowned for its anti-inflammatory, antioxidant, and immune-enhancing properties. In the realm of neurodegenerative diseases (NDDS), CLASP proteins, responsible for regulating microtubule dynamics in neurons, have emerged as critical players. Dysregulation of CLASP proteins is associated with NDDS, such as Alzheimer's, Parkinson's, and Huntington's diseases. Consequently, comprehending the role of CLASP proteins in NDDS holds promise for the development of innovative therapeutic interventions. The objectives of the research were to identify phytoconstituents in the hydroalcoholic extract of Uncaria tomentosa (HEUT), to evaluate its antioxidant potential through in vitro free radical scavenging assays and to explore its potential interaction with CLASP using in silico molecular docking studies. HPLC and LC-MS techniques were used to identify and quantify phytochemicals in HEUT. The antioxidant potential was assessed through DPPH, ferric reducing antioxidant power (FRAP), nitric oxide (NO) and superoxide (SO) free radical scavenging methods. Interactions between conventional quinovic acid, chlorogenic acid, epicatechin, corynoxeine, rhynchophylline and syringic acid and CLASP were studied through in silico molecular docking using Auto Dock 4.2. The HEUT extract demonstrated the highest concentration of quinovic acid derivatives. HEUT exhibited strong free radical-scavenging activity with IC50 values of 0.113 μg/ml (DPPH) and 9.51 μM (FRAP). It also suppressed NO production by 47.1 ± 0.37% at 40 μg/ml and inhibited 77.3 ± 0.69% of SO generation. Additionally, molecular docking revealed the potential interaction of quinovic acid with CLASP for NDDS. The strong antioxidant potential of HEUT and the interaction of quinovic acid with CLASP protein suggest a promising role in treating NDDS linked to CLASP protein dysregulation.

Photoprotective and antioxidant properties of scytonemin isolated from Antarctic cyanobacterium Nostoc commune Vaucher ex Bornet & Flahault and its potential as sunscreen ingredient

In the cosmetic industry there is an increasing demand for substances obtained from natural sources that can replace synthetic ones. Due to consumer demand for a protective filter with (SPF) labels in sunscreens, moisturizers, face make-up, and lipsticks worldwide, they produce tonnes of such products every year. Many species of cyanobacteria live in extreme environments, including sites with excessive doses of sunlight and drought. To survive in such extreme conditions, they produce compounds that allow both protection against ultraviolet radiation (UV), as well as the substances that are responsible for reducing oxidative stress. The aim of this study was to isolate, identify, and test the biological potential of the secondary metabolite scytonemin from the cyanobacterium Nostoc commune Vaucher ex Bornet et Flahault collected in Antarctica. The photoprotective effect was evaluated by the measurement of the sun protection factor (SPF) and the antioxidant activity was determined by two different assays including superoxide anion scavenging activity and free radical scavenging activity based on the amount of substance. An estimated SPF value of 33.34 ± 0.02 demonstrated that scytonemin might serve as a topically applicable ingredient for natural UV sunscreen cream.

Structural characterization, antioxidant and antimicrobial activities of polysaccharide from Akebia trifoliata (Thunb.) Koidz stem

Polysaccharides have a variety of beneficial pharmacological impact on human health. Akebia trifoliata (Thunb.) Koidz. has promising development prospects as a food resource with medicinal value. The aim of this study was to investigate the structural characterization, antioxidant, and antibacterial properties of A. trifoliata (Thunb.) Koidz polysaccharides (ATKPs). ATKP-II was purified from ATKP by DEAE-cellulose column with NaCl solution as eluent. ATKP and ATKP-II structures were characterized by high performance gel permeation chromatography, gas chromatography, ultraviolet-visible, Fourier transform infrared spectroscopy, thermogravimetry analysis and scanning electron microscopy. ATKP and ATKP-II were primarily composed of rhamnose, arabinose, xylose, mannose, glucose, and galactose in a molar percent of 1.6: 22.1: 3.6: 6.3: 55.7: 10.7, and 0.5: 22.1: 3.7: 10.2: 42.1: 21.4, respectively. Their structure may contain β-D-glucopyranose. The thermogravimetry analysis showed that ATKP and ATKP-II have good thermal stability at 230 °C and 200 °C, respectively. ATKP had the best antioxidant activities for 2, 2-diphenyl-1-picrylhydrazyl, hydroxyl, and superoxide free radical scavenging activities in vitro, and reducing ability than that of the purified polysaccharides. Moreover, ATKP was demonstrated an appreciable in vitro antibacterial activity, against Staphylococcus aureus, Bacillus subtilis, Salmonella, Penicillium italicum, Rhizopus and Aspergillus niger, but showed no activity against Escherichia coli and Saccharomycetes. These results demonstrated that ATKP displayed excellent antioxidant and antibacterial activities. This study provides a basis for the development and utilization in ATKP.

In Vitro Antioxidant and Fibroblast Migration Activities of Fractions Eluded from Dichloromethane Leaf Extract of Marantodes pumilum.

(1) The complexity of diabetes and diabetic wound healing remains a therapeutic challenge because proper and systematic wound care and management are essential to prevent chronic microbial infection and mechanical damage to the skin. Marantodes pumilum, locally known as 'Kacip Fatimah', is an herb that has been previously reported to possess anti-inflammatory, analgesic, antinociceptive and antipyretic properties. The current study aims to assess the antioxidant and fibroblast cell migration activities of the fractions eluded from the dichloromethane extract of M. pumilum leaves. (2) The total antioxidant capacity of M. pumilum was assessed using the total proanthocyanidins and phosphomolybdenum assays, while DPPH, nitric oxide, hydrogen peroxide and superoxide free radical scavenging assays were tested to determine the antioxidant potential of M. pumilum. An in vitro scratch wound assay was performed to measure the fibroblast cell migration rate using normal and insulin-resistant human dermal fibroblast cells. (3) All M. pumilum fractions exhibited good antioxidant and fibroblast cell migration activity, among which fractions A and E displayed the greatest effect. (4) M. pumilum's fibroblast migration activity could be attributed to its strong antioxidant properties along with its previously reported properties.

Screening and Inhibition Mechanism of Xanthine Oxidase Inhibitors in Ethanolic Extracts of Chimonanthus salicifolius Hu Leaves

This study aimed to evaluate the inhibition of the ethanol elutions of Chimonanthus salicifolius Hu leaves (CsHL) against xanthine oxidase (XO). The results of XO inhibition assay and enzymatic superoxide free radical scavenging assay in vitro showed that 70 % ethanol eluate (EE) had the best inhibitory effect and followed by 40 % EE. High performance liquid chromatograph analysis showed that quercetin and kaempferol were the potential active components of XO inhibition. The inhibition mechanism of quercetin and kaempferol on XO was investigated by kinetic analysis and fluorescence quenching titration assay. The molecular simulation further revealed that quercetin and kaempferol bind to XO mainly by hydrogen bonding and van der Waals, blocking the entry of substrates and leading to the inhibition of XO. In conclusion, the CsHL have inhibitory effects on XO activity, which provides a theoretical basis for relieving or preventing hyperuricemia and gout as a natural food or medicinal plant in the future.

Anticancer, Antioxidant Activity and Molecular Docking Studies of Saccharumoside-B

Objectives: The present study reports antioxidant potential, the in-vitro antiproliferative activity of saccharumoside-B, and molecular docking studies on the binding affinity of saccharumoside-B towards various proteins involved in breast cancer pathogenesis. Methods: The in-vitro antioxidant activity of saccharumoside-B was determined by DPPH, superoxide and nitric oxide free radical scavenging assays. The antiproliferative activity was evaluated in-vitro by MTT assay. The binding affinity of saccharumoside-B towards the HSP90, HER2, human estrogen receptor and tyrosine-protein kinase C-SRC were determined by the molecular docking studies. Results: Saccharumoside-B showed a significant dose-dependent antioxidant activity and potent dose-dependent antiproliferative effect on the MCF-7 breast cancer cell line (IC50 = 22.57±0.39µM) among all other cell lines studied. Tamoxifen was used as a positive control for MCF-7 cell line (IC50 = 27.97±1.07µM). The IC50 of saccharumoside-B on normal MCF-10A cell line (IC50>1000µM) showed a promising safety profile, whereas tamoxifen’s IC50 on MCF-10A normal cell line was found to be 29.6 ± 0.84 µM. The molecular docking analysis revealed that saccharumoside-B was inserted into the active site pockets of all the tested proteins involved in the breast cancer pathogenesis with varying binding affinities. Conclusion: The antioxidant studies revealed the potential antioxidant effect of saccharumoside-B. The in-vitro and in-silico studies are indicating the safe and multiple target protein inhibitory potential of saccharumoside-B against breast cancer. This study suggests that saccharumoside-B can be developed into a new class of anticancer drugs with a high safety profile in the future.

Gallic acid a flavonoid isolated from Euphorbia hirta antagonizes gamma radiation induced radiotoxicity in lymphocytes in vitro.

The current study was executed to isolate and evaluate gallic acid from Euphorbia hirta for in vitro radioprotective potentials against gamma irradiation caused radiotoxicity in human lymphocytes. The defatted E.hirta plant material was treated to methanol extraction using the soxhlet device. Bioflavonoids were isolated from the E.hirta methanol extract using column chromatography. In human cells exhibited to gamma radiation, separated flavonoid gallic acid was examined for in vitro radioprotective potentials using the micronucleus test, DNA fragmentation assay, superoxide free radical scavenging method, and apoptic assay. The frequency of micronuclei was considerably declined when cells were preprocessed with gallic acid (25g/mL) before being exhibited to 2Gy gamma radiation, as determined by the cytokinesis blocked micronucleus test. Similarly, pre-gamma radiation treatment of human cells with gallic acid led in markedly less DNA injury, as assessed by comet metrics like olive tail moment and percent tail DNA. Gallic acid (25g/mL) given to lymphocytes prior to gamma irradiation considerably decreased the percentage of apoptotic bodies. Gallic acid also considerably lowered the reactive oxygen species concentrations elicited by gamma radiation. Our findings showed that gallic acid protects lymphocytes isolated from human blood from gamma radiation-induced DNA destruction and anti-apoptotic activity, which could be because of inhibition of free radicals formed by gamma radiation as well as the decline of gamma radiation-induced oxidative stress.

Salidroside intensifies mitochondrial function of CoCl2-damaged HT22 cells by stimulating PI3K-AKT-MAPK signaling pathway

BackgroundSalidroside (Sal), an active component from Rhodiola crenulata, has been confirmed to exert neuroprotective effects against hypoxia. However, its molecular mechanisms of intensifying mitochondrial function still largely unknown. In the present study, we aimed to explore the mechanisms by which Sal heightened mitochondrial function in CoCl2-induced HT22 hypoxic injury. MethodsThe hypoxic condition of HT22 cells was performed by CoCl2 stimulus. We then investigated the effects of Sal on the viability of hypoxic HT22 cells by cell counting kit-8. The contents of lactate dehydrogenase (LDH) release in cultured supernatant were detected by using commercial biochemical kit. Superoxide free radical scavenging activity, total antioxidant capacity assay kit with ferric reducing ability of plasma and 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) methods were employed to detect the free radical scavenging ability and antioxidant capacity of Sal. Meanwhile, intracellular reactive oxygen species (ROS), Ca2+ and mitochondrial membrane potential (MMP) were determined by corresponding specific labeled probes. Mitochondrial morphology was tested by Mito-tracker green with confocal microscopy. Hoechst 33342 and Annexin V-FITC/propidium iodide staining were also employed to evaluate the effect of Sal on cell apoptosis. Oxygen consumption rate (OCR), real-time ATP production and proton efflux rate were measured using a Seahorse analyzer. Additionally, the potential interactions of Sal with PI3K-AKT signaling pathway-related proteins were predicted and tested by molecular docking, molecular dynamics simulation (MDS) and localized surface plasmon resonance (LSPR) techniques, respectively. Furthermore, the protein levels of p-PI3K, PI3K, p-AKT, AKT, p-JNK, JNK, p-p38 and p38 were estimated by western blot analysis. ResultsSal alleviated CoCl2-induced hypoxic injury in HT22 cells as evidenced by increased cell viability and decreased LDH release. In vitro antioxidant test confirmed that Sal had marvelous antioxidant abilities. The protected mitochondrial function by Sal treatment was illustrated by the decrease of ROS, Ca2+, mitochondrial fragment and the increase of MMP. In addition, Sal ameliorated the apoptosis of HT22 cells by decreasing Hoechst 33342 positive cells and the rate of apoptotic cells. Enhancement of energy metabolism in HT22 by Sal was demonstrated by increased OCR, real-time ATP generation and proton efflux rate. The molecular docking confirmed the potential binding of Sal to PI3K, AKT and CaMK II proteins with calculated binding energy of -1.32, -4.21 and -4.38 kcal/mol, respectively. The MDS test revealed the average hydrogen bond of complex Sal-PI3K and Sal-AKT were 0.79 and 4.46, respectively. The results of LSPR verified the potential binding of Sal to proteins PI3K, AKT and HIF-1α with affinity values of 5.20 × 10 − 3, 2.83 × 10 − 3 and 3.97 × 10 − 3 KD, respectively. Western blot analysis further argued that Sal consolidated the levels of p-PI3K and p-AKT. Meanwhile, Sal could downregulate the proteins expression of p-JNK and p-p38. ConclusionCollectively, our findings suggested that Sal can intensify mitochondrial function of CoCl2-simulated hypoxia injury in HT22 cells by stimulating PI3K-AKT-MAPK signaling pathway. Sal is a potential agent for mitochondrial protection against hypoxia with the underlying molecular mechanisms of energy metabolism being further elucidated.

A Comparative Study of Isolated Secondary Metabolites from Lichens and Their Antioxidative Properties.

Free radicals play a critical role in the chemical processes that occur in all cells. Pharmaceutical companies manufacture a variety of synthetically prepared antioxidants, but it is known that many of these can be carcinogenic. As a result, efforts are being made to find natural antioxidants that do not have these side effects. Lichens may be suitable candidates because they contain secondary metabolites with proven antioxidant properties. This could be explained by the presence of compounds with phenolic groups in lichens. The radical scavenging reaction is a chemical reaction governed by stoichiometry, and our aim is to determine the efficacy of these reactions. The aim of this study is to compare metabolite activity based on the same amount of substance involved in radical scavenging, calculated in micromoles rather than weight concentration. This provides an accurate way of comparing radical scavenging activity. We tested superoxide anion scavenging activity and free radical scavenging activity of isolated lichen secondary metabolites and their mixtures in different ratios. The following compounds were isolated and tested for antioxidant activity: gyrophoric acid (Umbilicaria hirsuta), evernic acid (Evernia prunastri), physodic acid, 3-hydroxyphysodic acid, physodalic acid and atranorin (Hypogymnia physodes), and usnic acid (as a synthetic compound). Of all the tested compounds, 3-hydroxyphysodic acid, as well as mixtures containing this metabolite, showed the strongest scavenging activity. The results also demonstrated that calculation by amount of substance leads to a new consideration of antioxidant activity.

Antioxidative, antimicrobial and anti-inflammatory activities and release of ultra-filtered antioxidative and antimicrobial peptides during fermentation of sheep milk: In-vitro, in-silico and molecular interaction studies

The antioxidative, antimicrobial, and anti-inflammatory properties of fermented sheep milk with L. fermentum (KGL4), as well as the generation of antioxidative and antimicrobial peptides, are evaluated in the study. Antioxidative and antibacterial activities in sheep milk fermented with KGL4 increased with incubation hours, along with varied antioxidative properties (ABTS assay: 35.12%, hydroxyl free radical scavenging assay: 29.12%, superoxide free radical scavenging activity: 36.38%). Antimicrobial activity of fermented sheep milk against E. faecalis (19 mm), S. typhimurium (15.67 mm), B. cereus (14 mm), and E. coli (13 mm) was also observed. Furthermore, after 48 h, the KGL4 showed maximum proteolysis (10.40 mg/ml) at 2.5% rate of addition sheep milk. The antioxidative and antimicrobial activities of fermented sheep milk fractions (3 kDa and 10 kDa permeates and retentates) were also investigated. The highest ABTS activity (26.90%) was found in 10 kDa permeate, whereas the 3 kDa retentate had higher hydroxyl free radical scavenging activity (69.20%) and 3 kDa permeate showed maximum superoxide free radical scavenging activity (32.85%). The 10 kDa retentate had shown maximum antimicrobial activity against S. typhimurium (13.67 mm) and E. faecalis (17.00 mm). Proteins spotted on 2D gel electrophoresis of KGL4 were varied from 10 to 70 kDa. RP-LC/MS was used to identify 5 novel peptide sequences from 2-D gel spots. Searches in the BIOPEP database confirmed the antioxidative and antimicrobial effects of the novel fermented sheep milk peptides. Fermented sheep milk with KGL4 (SMKGL4) significantly reduced excessive TNF-α, IL-6, & IL-1β production in LPS-activated RAW 264.7 cells.

Enhancing the In Vitro Biological Activity of Degraded Silk Sericin and Its Analog Metabolites.

Two sericins of high and low molecular weight (HS and LS) were prepared from commercial silkworm cocoon silk with a boiling water and Ca(OH)2 solution with ultrasonic treatments, respectively. This study first investigated the release concentration of the two abovementioned sericins in simulated saliva, gastric juice, and intestinal juice (pH 6.8, 2.0, and 7.4, respectively) within 10 h. The results showed that the order of sericin release rate and its amount in the simulated environment was gastric juice > saliva > intestinal juice. Second, the molecular weights of both sericin metabolites formed by in vitro enzymatic degradation were lower than 15 kDa. The α-glucosidase inhibitory activities of both sericins and their analog metabolites were positively correlated with their concentrations. The IC50 values of the HS- and LS-derived metabolites were 1.02 ± 0.12 mg/mL and 0.91 ± 0.15 mg/mL, respectively, which were five to seven times lower than those of both original sericins. The total antioxidant capacities and hydroxyl radical scavenging capacities of both metabolites were enhanced by one- to three-fold compared with HS and LS. These results indicate that both sericins, regardless of molecular size, have significantly enhanced antioxidant, superoxide free radical scavenging, and glycosidase inhibitory activities after simulated metabolism, and that LS is better than HS regardless of simulated digestion. These results confirm that sericin is important in the sustainable development and utilization of silk resources, especially the reduction in environmental pollution, and provides new ideas for the development of adjuvant treatments for diabetes and the development of foods with anti-hyperglycemic functions.

High-level production and extraction of C-phycocyanin from cyanobacteria Synechococcus sp. PCC7002 for antioxidation, antibacterial and lead adsorption

Global warming and climate change because carbon dioxide (CO2) release to atmosphere is the forecasting challenges to human being. We are facing how to overcome the dilemma on the balance between economic and environment, thus taking more efforts on green processes to meet agreement of sustainable society are urgent and crucial. The absorption of CO2 by microalgae reduces the impact of CO2 on the environment. In this study, the CO2 removal efficiency was the highest in the culture of Cyanobacterium Synechococcus sp. PCC7002 (also called blue-green algae), at 2% CO2 to reach a value of 0.86 g-CO2/g-DCW. The main product of PCC7002 is C-phycocyanin (C-PC) which regarding to phycobilisome complex in all cyanobacterial species. A 160% increasing C-PC was achieved in the cultivation under 100 μmol/m2/s light intensity, 12:12 light-period with 2% CO2 at 30 °C. The mix-culture of nitric and ammonia ions had positive effect on the cell growth and C-PC accumulation, thus realized the highest yield of 0.439 g-CPC/g-DCW. Additionally, the partial purified C-PC displayed 89% antioxidant activity of 2,2-diphenyl-1-picryhydrazyl (DPPH) and 11% of superoxide free radical scavenging activity, respectively. The production of C-PC from PCC7002 reduced the CO2 emission and exhibited antibacterial activity against Escherichia coli and lead ion adsorption at room temperature, which has the great potential for eco-friendly application.

Production and characterization of antioxidative peptides during lactic fermentation of goat milk

In the study, Lactobacillus fermentum M2 was characterized with potent antioxidative properties viz. ABTS assay (57.09%), HFRSA (57.30%), and SFRSA (51.40%) after 48 hr at 37°C in fermented goat milk. Further, the inoculation rate (2%) and incubation time (48 hr) for the production of peptides were also optimized using proteolytic activity at 37°C during the fermentation of goat milk. The molecular weight of proteins was analyzed by SDS-PAGE and protein bands in the range of 10–51 kDa were found. The 2D gel electrophoresis exhibited 41 protein spots between 10 and 51 kDa. The trypsin digested protein spots were injected into RPLC/MS for amino acid sequencing. The 3 and 10 kDa permeate fragments were injected into RPLC/MS for characterization of peptides, and the peptide masses from LC/MS were analyzed on MASCOT. Novel antioxidative peptides viz., SCQDQPTTLAR (2D-PAGE), TIDMESTEVFTKK (3 kDa Permeate), and YIQKEDVPSER (10 kDa Permeate) were identified from goat milk fermented with M2 by matching with BLASTp (NCBI), protein information resource (PIR), and BIOPEP databases. Practical applications Goat milk provides several health benefits, including antioxidative, antihypertensive, and antimicrobial properties. In terms of bio-functional characteristics, goat milk fermented with healthy lactic acid bacteria could even add more value. Goat milk fermented with Lactobacillus fermentum M2 exhibited promising bioactivities like hydroxyl free radical scavenging assay, superoxide free radical scavenging assay, antioxidative activity, and proteolytic activities. A few novel antioxidative peptides, viz., SCQDQPTTLAR, TIDMESTEVFTKK, and YIQKEDVPSER, were identified from goat milk fermented with M2 by similarity searches using BLASTp, PIR, and BIOPEP databases. Furthermore, L. fermentum M2 could be used for the production of fermented goat milk with antioxidative activity and bioactive peptides.

Screening of in vitro free radical scavenging activities of the fruit extracts of Psidium guajava L.

The objective of the current investigation was to evaluate the in vitro free radical scavenging properties of fruit extracts of two varieties of Psidium guajava L. (PG) i.e., Lalit (LA) and Allahabad safeda (AS). The ethanolic fruit extracts of LA and AS of PG were obtained from the soxhlet extraction procedure and the same was used for in vitro antioxidant assays. Total phenols, flavonoid contents and free radical scavenging assays viz., ABTS, DPPH, nitric oxide and superoxide were analysed at 100, 200, 300, 400, 500, 600 and 700 µg/mL concentrations. Phytochemical investigation exhibited higher phenols, flavonoid contents in fruit extracts of LA compared to AS. The fruit extracts of PG showed remarkable ABTS, DPPH, nitric oxide and superoxide free radical scavenging activity in a dose dependent manner. The fruit extracts of LA exhibited higher free radical scavenging activity in contrast to AS. The present investigation revealed, a high in vitro antioxidant property of LA due to its higher phenolic content. Hence, the fruit extract of Psidium guajava L. can be used in therapeutics for the treatment of pathologic conditions involving free radicals.

Comparing Glycerol with Dimethylformamide and Combination for Cryopreservation of Pug Dog Semen in Egg Yolk Plasma Containing Extender

Background: In order to find an alternative cryoprotectant for canine semen, different cryoprotectants, as dimethyl sulphoxide and ethylene glycol, dimethyle formamide either alone or in combination with glycerol have been evaluated on pooled semen of different breeds with different opinions. Methods: In the current study the effect of glycerol, DMF and their combination in Tris-citric acid-fructose egg yolk plasma extender on motility, viability, plasma membrane integrity (PMI), acrosome integrity (AI), inner mitochondrial membrane potential (IMMP), reactive oxygen species (ROS) and antioxidant enzymes in frozen-thaw semen of pug breed was compared. Result: Values for motility, viability, (PMI), AI were significantly (p less than 0.05) high in TCFEYP-G compared to TCFEYP-DMF and TCFEYP-G+DMF at post thaw. However, HIMMP and MIMMP/MDA were non-significantly (P greater than 0.05) high and low in TCFEYP-G compared to TCFEYP-DMF and TCFEYP- G + DMF at post thaw, respectively. MDA concentration was significantly (p less than 0.05) low in TCFEYP-G compared to TCFEYP-DMF and TCFEYP-G + TCFEYP-DMF extenders. However, H2O2, scavenging capacity of spermatozoa, cryopreserved in TCFEYP-G was significantly (p less than 0.05) higher than in TCFEYP-DMF and TCFEYP-G + TCFEYP-DMF. However, there was no significant (p greater than 0.05) difference in superoxide free radical scavenging activity and nitrite concentration of spermatozoa among the three extenders. Activity of GPX, SOD and Catalase was also significantly (p less than 0.05) higher in TCFEYP-G compared to TCFEYP-DMF and TCFEYP-G+DMF. However, values were reverse for GRE activity. This study concludes that glycerol is a healthier cryoprotectant to protect canine spermatozoa from cryo-injury during freezing-thawing process.

Evaluation of Antioxidant, Immunomodulatory and Anticancer Properties of Methanolic Extract of Neolamarckia cadamba Linn. Fruits

Abstract: Background: The immune system plays an important role in various diseased conditions. Patients with compromised immune function are at increased risk of exposure. Thus, the evaluation of immunomodulatory activity of plant extracts may be helpful for prophylactic treatment approach. Objectives: The aim of present study is to evaluate the effect of methanolic extract of Neolamarckia cadamba Linn. fruits for antioxidant, immunomodulatory and anticancer properties using in-vitro and in-vitro models. Methods: In the present investigation, antioxidant property of methanolic extract of Neolamarckia cadamba fruits (NCM) was evaluated by superoxide radical and DPPH scavenging activity. The immunomodulatory property of NCM extract was analysed by determining cytokines release (IL-2 and TNFα) in murine primary thymocytes. The effects of NCM extract on humoral and cellular immunity was analysed under in vivo conditions. The anticancer activity of the extract was evaluated on Jurkat J6 cell line by MTT assay. Results: The methanolic extract of Neolamarckia cadamba fruits showed superoxide and DPPH free radical scavenging antioxidant activity and significantly increased the cytokines release (IL-2 and TNFα) in murine primary thymocytes. The increased hemagglutination antibody titre and neutrophil adhesion as well as the restoration of cyclophosphamideinduced myelosuppression and DTH response confirm the stimulatory effect of NCM on T cells. In-vitro results on Jurkat J6 cell line showed the significant increase in cell death confirming its anticancer activity. Conclusion: Thus, it can be concluded that the methanolic extract of the fruits of Neolamarckia cadamba possesses antioxidant activity, immunomodulatory property as well as anticancer potential, suggesting its therapeutic efficiency in the treatment of immune disorders as well as cancer. Key words: Neolamarckia cadamba, Immunomodulatory, Antioxidative, Anticancer, Cytokines, Neutrophil adhesion.

Formulation Evaluation and In- Vitro Antioxidant activity of Microparticles of Syzygium Cumini Plant Extract

Sygygium cumini, commonly known as Malabar plum, Java plum, black plum, jamun or jambolan, is an evergreen tropical tree in the flowering plant family Myrtaceae. The present study the antioxidant effects of microparticles of S. cumini plant extract were evaluated using in vitro, 2, 2-diphenylpicrylhydrazine photometric assay, total antioxidant activity and super oxide free radical scavenging activity. The microparticles showed different levels of radicals scavenging activity in a dose-dependent manner over the range of 50–150 µg/mL concentration, indicating the high antioxidative capacity of the extract. In all the experimental screening models microparticles of aqueous extract show the maximum inhibition of free radicals comparison of other experimental model at different concentrations. These findings suggest that the seed of Syginium cumini has potent antioxidant activity which may be responsible for some of its reported pharmacological activities and can be used as antioxidant supplement.

Design, Synthesis and Biological Activities of Dihydroaurones

To widen aurones applicability in achromatic food and cosmetic applications, a series of dihydroaurones were designed to mimic natural aurones as well as synthetic aurones. Dihydroaurones have been synthesized from the corresponding aurones by hydrogenation. These dihydroaurones and their corresponding aurones were screened for antioxidant, anti-inflammatory and tyrosinase enzyme inhibitory activity. Synthesized dihydroaurones (3b-f) displayed superior antioxidant activity in superoxide free radical scavenging assay than the standard gallic acid. Dihydroaurones (3b-f) also exhibited significant tyrosinase enzyme inhibitory activity and two dihydroaurones (3h, 3j) showed promising 5-lipoxygenase inhibitory activity.

Effect of microwave-drying on the quality and antioxidant properties of Ganoderma lucidum fermented sea-buckthorn tea

Abstract Effects of microwave power on the sensory properties (taste and aroma), chemical composition (catechins, caffeine, crude protein, and amino acid), active composition (flavones, triterpene, polysaccharide, and ergosterol) and antioxidant properties (superoxide free radical and hydroxyl radical scavenging abilities, reducing power) of Ganoderma lucidum fermented sea-buckthorn tea were investigated. G. lucidum fermented sea-buckthorn tea was dehydrated using microwaves at three power settings: 125, 250, and 500. After microwave treatment, a statistically significant difference in the chemical composition, active composition and antioxidant capacity (p < 0.05) of teas dried at different power settings was found. These results indicate that 250 W microwave treatments could effectively release and activate active components, improving the antioxidant activity of fermented tea.