Search results for: cellular antioxidant activity

Authors: Vladimir Berezin, Andrey Bogoyavlenskiy, Pavel Alexyuk, Madina Alexyuk, Aizhan Turmagambetova, Irina Zaitseva, Nadezhda Sokolova, Elmira Omirtaeva

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Introduction: Triterpene saponins of plant origin are one of the most promising candidates for elaboration of novel adjuvants. Due to the combination of immunostimulating activity and the capacity interact with amphipathic molecules with formation of highly immunogenic nanocomplexes, triterpene saponins could serve as a good adjuvant/delivery system for vaccine use. In the research presented adjuvants on the base of nanocomplexes contained triterpene saponins isolated from Glycyrrhiza glabra and Saponaria officinalis plants indigenous to Kazakhstan were elaborated for influenza vaccine use. Methods: Purified triterpene saponins 'Glabilox' and 'SO1' with low toxicity and high immunostimulatory activity were isolated from plants Glycyrrhiza glabra L. and Saponaria officinalis L. by high-performance liquid chromatography (HPLC) and identified using electrospray ionization mass spectrometry (ESI-MS). Influenza virus A/St-Petersburg/5/09 (H1N1) propagated in 9-days old chicken embryos was concentrated and purified by centrifugation in sucrose gradient. Nanocomplexes contained lipids, and triterpene saponins Glabilox or SO1 were prepared by dialysis technique. Immunostimulating activity of experimental vaccine preparations was studied in vaccination/challenge experiments in mice. Results: Humoral and cellular immune responses and protection against influenza virus infection were examined after single subcutaneous and intranasal immunization. Mice were immunized subunit influenza vaccine (HA+NA) or whole virus inactivated influenza vaccine in doses 3.0/5.0/10.0 µg antigen/animal mixed with adjuvant in dose 15.0 µg/animal. Sera were taken 14-21 days following single immunization and mice challenged by A/St-Petersburg/5/09 influenza virus in dose 100 EID₅₀. Study of experimental influenza vaccine preparations in animal immunization experiments has shown that subcutaneous and intranasal immunization with subunit influenza vaccine mixed with nanocomplexes contained Glabilox or SO1 saponins stimulated high levels of humoral immune response (IgM, IgA, IgG1, IgG2a, and IgG2b antibody) and cellular immune response (IL-2, IL-4, IL-10, and IFN-γ cytokines) and resulted 80-90% protection against lethal influenza infection. Also, single intranasal and single subcutaneous immunization with whole virus inactivated influenza vaccine mixed with nanoparticulated adjuvants stimulated high levels of humoral and cellular immune responses and provided 100% protection against lethal influenza infection. Conclusion: The results of study have shown that nanocomplexes contained purified triterpene saponins Glabilox and SO1 isolated from plants indigenous to Kazakhstan can stimulate a broad spectrum of humoral and cellular immune responses and induce protection against lethal influenza infection. Both elaborated adjuvants are promising for incorporation to influenza vaccine intended for subcutaneous and intranasal routes of immunization.

Keywords: influenza vaccine, adjuvants, triterpene saponins, immunostimulating activity

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Authors: Shahira M. Ezzat, Marwa I. Ezzat, Mona M. Okba, Esther T. Menze, Ashraf B. Abdel-Naim, Shahnas O. Mohamed

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Background: In order to decrease the burden of the high cost of synthetic drugs, it is important to focus on phytopharmaceuticals. The aim of our study was to search for the mechanism of ginger (Zingiber officinale Roscoe) anti-inflammatory potential and to correlate it to its biophytochemicals. Methods: Various extracts viz. water, 50%, 70%, 80%, and 90% ethanol were prepared from ginger rhizomes. Fractionation of the aqueous extract (AE) was accomplished using Diaion HP-20. In vitro anti-inflammatory activity of the different extracts and isolated compounds was evaluated by protein denaturation inhibition, membrane stabilization, protease inhibition, and anti-lipoxygenase assays. In vivo anti-inflammatory activity of AE was estimated by assessment of rat paw oedema after carrageenan injection. Prostaglandin E2 (PGE2), certain inflammation markers (TNF-α, IL-6, IL-1α, IL-1β, INFr, MCP-1MIP, RANTES, and Nox) levels and MPO activity in the paw edema exudates were measured. Total antioxidant capacity (TAC) was also determined. Histopathological alterations of paw tissues were scored. Results: All the tested extracts showed significant (p < 0.1) anti-inflammatory activities. The highest percentage of heat induced albumin denaturation (66%) was exhibited by the 50% ethanol (250 μg/ml). The 70 and 90% ethanol extracts (500 μg/ml) were more potent as membrane stabilizers (34.5 and 37%, respectively) than diclofenac (33%). The 80 and 90% ethanol extracts (500 μg/ml) showed maximum protease inhibition (56%). The strongest anti-lipoxygenase activity was observed for the AE. It showed more significant lipoxygenase inhibition activity than that of diclofenac (58% and 52%, respectively) at the same concentration (125 μg/ml). Fractionation of AE yielded four main fractions (Fr I-IV) which showed significant in vitro anti-inflammatory. Purification of Fr-III and IV led to the isolation of 6-poradol (G1), 6-shogaol (G2); methyl 6- gingerol (G3), 5-gingerol (G4), 6-gingerol (G5), 8-gingerol (G6), 10-gingerol (G7), and 1-dehydro-6-gingerol (G8). G2 (62.5 ug/ml), G1 (250 ug/ml), and G8 (250 ug/ml) exhibited potent anti-inflammatory activity in all studied assays, while G4 and G5 exhibited moderate activity. In vivo administration of AE ameliorated rat paw oedema in a dose-dependent manner. AE (at 200 mg/kg) showed significant reduction (60%) of PGE2 production. The AE at different doses (at 25-200 mg/kg) showed significant reduction in inflammatory markers except for IL-1α. AE (at 25 mg/kg) is superior to indomethacin in reduction of IL-1β. Treatment of animals with the AE (100, 200 mg/kg) or indomethacin (10 mg/kg) showed significant reduction in TNF-α, IL-6, MCP-1, and RANTES levels, and MPO activity by about (31, 57 and 32% ) (65, 60 and 57%) (27, 41 and 28%) (23, 32 and 23%) (66, 67 and 67%) respectively. AE at 100 and 200 mg/kg was equipotent to indomethacin in reduction of NOₓ level and in increasing the TAC. Histopathological examination revealed very few inflammatory cells infiltration and oedema after administration of AE (200 mg/kg) prior to carrageenan. Conclusion: Ginger anti-inflammatory activity is mediated by inhibiting macrophage and neutrophils activation as well as negatively affecting monocyte and leukocyte migration. Moreover, it produced dose-dependent decrease in pro-inflammatory cytokines and chemokines and replenished the total antioxidant capacity. We strongly recommend future investigations of ginger in the potential signal transduction pathways.

Keywords: anti-lipoxygenase activity, inflammatory markers, 1-dehydro-6-gingerol, 6-shogaol

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Authors: Jelena Vulić, Jasna Čanadanović-Brunet, Gordana Ćetković, Sonja Djilas, Vesna Tumbas Šaponjac

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Caneberries (raspberries and blackberries) are among the most popular berries in the world, which are consumed as fresh and processed to juice, jams, confitures and other products or as ingredients for different foods. These fruits are known as a rich source of phenolic compounds such as phenolic acids and anthocyanins. Antioxidant activity (AA) of caneberry juices was improved by addition of phenolic compounds which were extracted from two raspberry cultivars (Rubus idaeus, cv. 'Willamette' (RW) and 'Meeker' (RM)) and two blackberry cultivars (Rubus fruticosus, cv. 'Čačanka' (BC) and 'Thornfree' (BT)) pomace, a by-product in juice processing. The total phenolic contents in raspberry and blackberry pomace extracts were determined spectrophotometrically using the Folin-Ciocalteu reagens. The phenolic concentrations in caneberries (RW, RM, BC and BT) pomace extracts were 43.67 ± 2.13 mg GAE/g, 26.25 ± 1.18 mg GAE/g, 46.01 ± 3.26 mg GAE/g and 61.59 ± 1.14 mg GAE/g, respectively. In order to obtain enriched juices, phenolic compounds were applied at concentration of 0.05 mg GAE/ 100 ml. Antioxidant activities of caneberry juices and caneberry enriched juices were measured using stable 1.1-diphenyl-2-picrylhydrazyl (DPPH) radicals. AADPPH of RW, RM, BC and BT juices and enriched juices with addition of 0.01 µg GAE/ml, changed from 37.12% to 93.01%, 23.26% to 91.57%, 53.61% to 95.65% and 52.06% to 93.13%, respectively, while IC50 values of RW, RM, BC and BT juices and enriched juices were diminished 6.33, 19.00, 6.33 and 4.75 times, respectively. Based on the obtained results it can be concluded that phenolic enriched juices were significantly more effective on DPPH radicals. Caneberry juices enriched with waste material are a good source of natural pigments and antioxidants and could be used as functional foods.

Keywords: caneberry, enriched juice, phenolic antioxidant, DPPH radical

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Authors: Aman Upaganlawar, Chandrasekhar Upasani

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The present study was designed to screen the neuroprotective and antioxidant activity of corosolic acid in painful diabetic neuropathy (DN). Diabetes was induced in rats by single dose of STZ (60mg/kg, i.p). Diabetic rats were tested every week for the development of pain, at 5th week rats showed sensation of pain. At 6th week the rats developed significant neuropathic pain. They were divided into different groups and treated with Corosolic acid (2 and 4 mg/kg, p.o) for further two weeks. Pain was assessed in the diabetic rats by mechano-tactil allodynia, mechanical hyperalgesia and cold allodynia. At the end of treatment period rats were scarified and biochemical changes such as plasma glucose level, endogenous antioxidants (Lipid peroxidation, reduced glutathione, superoxide dismutase and catalase) in sciatic nerve were evaluated. Further Na+/K+ ATPase and nitric oxide content was also evaluated. Treatment with corosolic acid for two weeks restored the altered body weight and elevated blood sugar level. Further corosolic acid showed dose dependent reduction in pain in neuropathic animals. The level of endogenous antioxidants enzymes, Na+/K+ ATPase and nitric oxide were significantly prevented. In conclusion, the result of the present study suggests the antidiabetic, antioxidant and neuroprotectieve property of corosolic acid in diabetic rats with neuropathic pain.

Keywords: neuropathic pain, diabetes, corosolic acid, antioxidant

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Authors: Shwu-Ling Peng, Chiung-Man Tsai, Chia-Jui Weng

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Chronic micro-inflammation is a hallmark of many aging-related neurodegenerative and metabolic syndrome-driven diseases. In high glucose (HG) environment, reactive oxygen species (ROS) is generated and the ROS induced inflammation, cytokines secretion, DNA damage, and cell cycle arrest to lead to cellular senescence. Water chestnut shell (WCS) is a plant hull which containing polyphenolic compounds and showed antioxidant and anticancer activities. Orchid, which containing a natural polysaccharide compound, possesses many physiological activities including anti-inflammatory and neuroprotective effects. These agricultural plants might be able to reduce oxidative stress and inflammation. This study was used HG-induced human normal dermal fibroblasts (HG-HNDFs) as an in vitro model to disclose the effects of water extract of Phalaenopsis orchid flower (WEPF) and ethanol extract of water chestnut shell (EEWCS) on the anti-aging and their underlying molecular mechanisms. The toxicity of extracts on human normal dermal fibroblasts (HNDFs) was determined by MTT method. The senescence of cells was assayed by β-galactosidase (SA-β-gal) kit. ROS and nitrate production was analyzed by Intracellular ROS contents and ELISA, respectively. Western blotting was used to detect the proteins in cells. The results showed that the exposure of HNDFs to HG (30 mM) for 72 h were caused cellular senescence and arrested cells at G0/G1 phase. Indeed, the treatment of HG-HNDFs with WEPF (200 μg/ml) and EEWCS (10 μg/ml) significantly released cell cycle arrest and promoted cell proliferation. The G1/S phase transition regulatory proteins such as protein retinoblastoma (pRb), p53, and p16ᴵᴺᴷ⁴ᵃ depressed by WEPF and EEWCS were also observed. Additionally, the treatment of WEPF and EEWCS increased the activity of HO-1 through upregulating Nrf2 as well as decreased the ROS and NO of HG-HNDFs. Therefore, the senescence marker protein-30 (SMP30) in cells was diminished. In conclusion, the WEPF and EEWCS might inhibit HG-induced aging of HNDFs by reducing oxidative stress and free radicals.

Keywords: agricultural plant extract, anti-aging, high glucose, Phalaenopsis orchid flower, water chestnut shell

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Authors: D. M. El-Tanbouly, R. M. Abdelsalam, A. S. Attia, M. T. Abdel-Aziz

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Lipopolysaccharide (LPS) endotoxin, a component of the outer membrane of Gram-negative bacteria, is involved in the pathogenesis of sepsis. LPS administration induces systemic inflammation that mimics many of the initial clinical features of sepsis and has deleterious effects on several organs including the liver and eventually leading to septic shock and death. The present study aimed to investigate the protective effect of magnesium, a well-known cofactor in many enzymatic reactions and a critical component of the antioxidant system, on hepatic damage associated with LPS induced- endotoxima in mice. Mg (20 and 40 mg/kg, po) was administered for 7 consecutive days. Systemic inflammation was induced one hour after the last dose of Mg by a single dose of LPS (2 mg/kg, ip) and three hours thereafter plasma was separated, animals were sacrificed and their livers were isolated. LPS-treated mice suffered from hepatic dysfunction revealed by histological observation, elevation in plasma transaminases activities, C-reactive protein content and caspase-3, a critical marker of apoptosis. Liver inflammation was evident by elevation in liver cytokines contents (TNF-α and IL-10) and myeloperoxidase (MPO) activity. Additionally, oxidative stress was manifested by increased liver lipoperoxidation, glutathione depletion, elevated total nitrate/nitrite (NOx) content and glutathione peroxidase (GPx) activity. Pretreatment with Mg largely mitigated these alternations through its anti-inflammatory and antioxidant potentials. Mg, therefore, could be regarded as an effective strategy for prevention of liver damage associated with septicemia.

Keywords: LPS, liver damage, magnesium, septicemia

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Authors: Muhammad H. Alu’datt, Inteaz Alli

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This study was conducted to prepare and evaluate the biological properties of protein co-precipitates from flaxseed and soybean. Protein was prepared by NaOH extraction through the mixing of soybean flour (Sf) and flaxseed flour (Ff) or mixtures of soybean extract (Se) and flaxseed extract (Fe). The protein co-precipitates were precipitated by isoelectric (IEP) and isoelectric-heating (IEPH) co-precipitation techniques. Effects of extraction and co-precipitation techniques on co-precipitate yield were investigated. Native-PAGE, SDS-PAGE were used to study the molecular characterization. Content and antioxidant activity of extracted free and bound phenolic compounds were evaluated for protein co-precipitates. Removal of free and bound phenolic compounds from protein co-precipitates showed little effects on the electrophoretic behavior of the proteins or the protein subunits of protein co-precipitates. Results showed that he highest protein contents and yield were obtained in for Sf-Ff/IEP co-precipitate with values of 53.28 and 25.58% respectively as compared to protein isolates and other co-precipitates. Results revealed that the Sf-Ff/IEP showed a higher content of bound phenolic compounds (53.49% from total phenolic content) as compared to free phenolic compounds (46.51% from total phenolic content). Antioxidant activities of extracted bound phenolic compounds with and without heat treatment from Sf-Ff/IEHP were higher as compared to free phenolic compounds extracted from other protein co-precipitates (29.68 and 22.84%, respectively).

Keywords: antioxidant, phenol, protein co-precipitate, yield

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Authors: Jasna Čanadanović-Brunet, Lidija Jevrić, Gordana Ćetković, Vesna Tumbas Šaponjac, Jelena Vulić, Slađana Stajčić

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Wheat contains high amount of nutrients such as dietary fiber, resistant starch, vitamins, minerals and microconstituents, which are building blocks of body tissues, but also help in the prevention of diseases such as cardiovascular disease, cancer and diabetes. Sprouting enhances the nutritional value of whole wheat through biosynthesis of tocopherols, polyphenols and other valuable phytochemicals. Since the nutritional and sensory benefits of germination have been extensively documented, using of sprouted grains in food formulations is becoming a trend in healthy foods. The present work addressed the possibility of using freeze-dried sprouted wheat powder, obtained from spelt-wheat cv. ‘Nirvana’ (Triticum spelta L.) and winter wheat cv. ‘Simonida’ (Triticum aestivum L. ssp. vulgare var. lutescens), as a source of phytochemicals, to improve the functional status of the consumer. The phytochemicals' content (total polyphenols, flavonoids, chlorophylls and carotenoids) and biological activities (antioxidant activity on DPPH radicals and antiinflammatory activity) of sprouted wheat powders were assessed spectrophotometrically. The content of flavonoids (216.52 mg RE/100 g), carotenoids (22.84 mg β-carotene/100 g) and chlorophylls (131.23 mg/100 g), as well as antiinflammatory activity (EC50=3.70 mg/ml) was found to be higher in sprouted spelt-wheat powder, while total polyphenols (607.21 mg GAE/100 g) and antioxidant activity on DDPPH radicals (EC50=0.27 mmol TE/100 g) was found to be higher in sprouted winter wheat powders. Simulation of gastro-intestinal digestion of sprouted wheat powders clearly shows that intestinal digestion caused a higher release of polyphenols than gastric digestion for both samples, which indicates their higher bioavailability in the colon. The results of the current study have shown that wheat sprouts can provide a high content of phytochemicals and considerable bioactivities. Moreover, data reported show that they contain a unique pattern of bioactive molecules, which make these cereal sprouts attractive functional foods for a health-promoting diet.

Keywords: wheat, sprouts, phytochemicals, bioactivity

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Authors: Ana Jelenkovic, Nevena Kardum, Vuk Stevanovic, Ivana Šarac, Kristina Dmitrovic, Stevan Stevanovic, Maria Glibetic

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Physical activity is well known for its beneficial health implications, however, excess oxygen consumption may impair oxidative status of the cell and affect membrane fatty acid (FA) composition. Polyphenols are well-established antioxidants, which can incorporate in cell membranes and protect them from oxidation. Therefore, our aim was to investigate how an 8-week aerobic training alters erythrocyte FA composition and activities of enzymes (superoxide dismutase, glutathione peroxidase and catalase), and to what extent polyphenol-rich Aronia juice (AJ) counteracts these potential alterations. We included 28 healthy women aged 19-29, with mean body mass index (BMI) of 21.2±2.7kg/m² and assigned them into three groups. The first group performed 1 hour of aerobic training three times per week (T); the second group trained in the same way and received 100 ml/day AJ as a part of their regular diet (TAJ), while the third group was the control one (C). Study analyses were performed at baseline and at the end of the intervention and included: anthropometric and biochemical measurements, determination of erythrocyte FA profile with gas-liquid chromatography and determination of enzymes’ activity with spectrophotometry. Statistical analyses were carried out with SPSS 20.0, with p < 0.05 considered as significant. The paired t-test revealed a significant decrease in the saturated FA content and in ω6/ω3 ratio in TAJ group. Furthermore, ω3 and docosahexaenoic acid (DHA) content increased, as well as the percentage of polyunsaturated FA and unsaturation index, which clearly pointed out that AJ supplementation with aerobic training protected cellular membranes from lipid peroxidation. No significant changes were observed in the two other groups. The between-group comparisons (ANCOVA) confirmed the synergistic effect of AJ supplementation and physical activity: DHA and ω3 contents were much higher, while ω6/ω3 ratio was significantly lower in the TAJ group compared with C. We also found that after the 8 weeks period, participants in TAJ group had a higher unsaturation index and lower saturated FA concentration than subjects from T group, suggesting that AJ polyphenols might be involved in that particular pathway. We found no significant changes in enzymes’ activities apart from a significantly higher superoxide dismutase activity in T group compared with the other two groups. Our results imply that supplementation with polyphenol-rich AJ may prevent membrane lipids from peroxidation in healthy subjects with regular aerobic activity.

Keywords: Aronia juice, aerobic training, fatty acids, oxidative status

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Authors: B. Voucko, M. Benkovic, N. Cukelj, S. Drakula, D. Novotni, S. Balbino, D. Curic

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Oxidative stress is considered as one of the causes leading to metabolic disorders in humans. Therefore, the ability of antioxidants to inhibit free radical production is their primary role in the human organism. Antioxidants originating from cereals, especially flavonoids and polyphenols, are mostly bound and indigestible. Micronization damages the cell wall which consecutively results in bioactive material to be more accessible in vivo. In order to ensure complete fragmentation, micronization is often combined with high temperatures (e.g., for bran 200°C) which can lead to degradation of bioactive compounds. The innovative non-thermal technology of cryo-milling is an ultra-fine micronization method that uses liquid nitrogen (LN2) at a temperature of 195°C to freeze and cool the sample during milling. Freezing at such low temperatures causes the material to become brittle which ensures the generation of fine particles while preserving the bioactive content of the material. The aim of this research was to determine if production of ultra-fine material with cryo-milling will result in the augmentation of available bioactive compounds of buckwheat and pumpkin seed cake. For that reason, buckwheat and pumpkin seed cake were ground in a ball mill (CryoMill, Retch, Germany) with and without the use of LN2 for 8 minutes, in a 50 mL stainless steel jar containing one grinding ball (Ø 25 mm) at an oscillation frequency of 30 Hz. The cryo-milled samples were cooled with LN2 for 2 minutes prior to milling, followed by the first cycle of milling (4 minutes), intermediary cooling (2 minutes), and finally the second cycle of milling (further 4 minutes). A continuous process of milling was applied to the samples ground without freezing with LN2. Particle size distribution was determined using the Scirocco 2000 dry dispersion unit (Malvern Instruments, UK). Antioxidant activity was determined by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) test and ferric reducing antioxidant power (FRAP) assay, while the total phenol content was determined using the Folin Ciocalteu method, using the ultraviolet-visible spectrophotometer (Specord 50 Plus, Germany). The content of the free phenolic acids, rutin in buckwheat, tyrosol in pumpkin seed cake, was determined with an HPLC-PDA method (Agilent 1200 series, Germany). Cryo-milling resulted in 11 times smaller size of buckwheat particles, and 3 times smaller size of pumpkin seed particles than milling without the use of LN2, but also, a lower uniformity of the particle size distribution. Lack of freezing during milling of pumpkin seed cake caused a formation of agglomerates due to its high-fat content (21 %). Cryo-milling caused augmentation of buckwheat flour antioxidant activity measured by DPPH test (23,9%) and an increase in available rutin content (14,5%). Also, it resulted in an augmentation of the total phenol content (36,9%) and available tyrosol content (12,5%) of pumpkin seed cake. Antioxidant activity measured with the FRAP test, as well as the content of phenolic acids remained unchanged independent of the milling process. The results of this study showed the potential of cryo-milling for complete raw material utilization in the food industry, as well as a tool for extraction of aimed bioactive components.

Keywords: bioactive, ball-mill, buckwheat, cryo-milling, pumpkin seed cake

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Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

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One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures

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Authors: Ripon Sarkar, Kabita Chaterjee, Ananya Barui

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Smoking is one of the leading causes of oral cancer. Cigarette smoke affects various cellular parameters and alters molecular metabolism of cells. Epithelial cells losses their cytoskeleton structure, membrane integrity, cellular polarity that subsequently initiates the process of epithelial cells to mesenchymal transition due to long exposure of cigarette smoking. It changes the normal cellular metabolic activity which induces oxidative stress and enhances the reactive oxygen spices (ROS) formation. Excessive ROS and associated oxidative stress are considered to be a driving force in alteration in cellular phenotypes, polarity distribution and mitochondrial metabolism. Noninvasive assessment of such parameters plays essential role in development of routine screening system for early diagnosis of oral cancer. Electrical cell-substrate impedance sensing (ECIS) is one of such method applied for detection of cellular membrane impedance which can be correlated to cell membrane integrity. Present study intends to explore the alteration in cellular impedance along with the expression of cellular polarity molecules and cytoskeleton distributions in oral epithelial cells of habitual smokers and to correlate the outcome to that of clinically diagnosed oral leukoplakia and oral squamous cell carcinoma patients. Total 80 subjects were categorized into four study groups: nonsmoker (NS), cigarette smoker (CS), oral leukoplakia (OLPK) and oral squamous cell carcinoma (OSCC). Cytoskeleton distribution was analyzed by staining of actin filament and generation of ROS was measured using assay kit using standard protocol. Cell impedance was measured through ECIS method at different frequencies. Expression of E-cadherin and protease-activated receptor (PAR) proteins were observed through immune-fluorescence method. Distribution of actin filament is well organized in NS group however; distribution pattern was grossly varied in CS, OLPK and OSCC. Generation of ROS was low in NS which subsequently increased towards OSCC. Expressions of E-cadherin and change in cellular electrical impedance in different study groups indicated the hallmark of cancer progression from NS to OSCC. Expressions of E-cadherin, PAR protein, and cell impedance were decreased from NS to CS and farther OSCC. Generally, the oral epithelial cells exhibit apico-basal polarity however with cancer progression these cells lose their characteristic polarity distribution. In this study expression of polarity molecule and ECIS observation indicates such altered pattern of polarity among smoker group. Overall the present study monitored the alterations in intracellular ROS generation and cell metabolic function, membrane integrity in oral epithelial cells in cigarette smokers. Present study thus has clinical significance, and it may help in developing a noninvasive technique for early diagnosis of oral cancer amongst susceptible individuals.

Keywords: cigarette smoking, early oral cancer detection, electric cell-substrate impedance sensing, noninvasive screening

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Authors: F. T. Senberber, N. Tugrul, E. Moroydor Derun

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As a nutraceutical, sumac (Rhus Coriaria) was extracted by using different solvents of methanol, ethanol, and water. The DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) method of free radical scavenging capacity was used to determine the effects of solvent on antioxidant activities of the plant. The total phenolic content was studied by The Folin Ciocalteu Reagent method. The antioxidant activities of extracts exhibit minor changes in different solvents and varied in the range of 84.3–86.4 %. The total phenolic contents are affected by the selected solvent. The highest total phenolic content was determined at the liquid phase of water and it was estimated as 26.3 mg/g in gallic acid.

Keywords: DPPH, solvent, sumac, total phenolic content

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Authors: Muhammad Ashfaq

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Main theme: This prime focus of this study is on the role of digital technology in crime prevention, with special focus on Cellular Forensic Unit, Capital City Police Peshawar-Khyber Pakhtunkhwa-Pakistan. Objective(s) of the study: The prime objective of this study is to provide statistics, strategies and pattern of analysis used for crime prevention in Cellular Forensic Unit of Capital City Police Peshawar, Khyber Pakhtunkhwa-Pakistan. Research Method and Procedure: Qualitative method of research has been used in the study for obtaining secondary data from research wing and Information Technology (IT) section of Peshawar police. Content analysis was the method used for the conduction of the study. This study is delimited to Capital City Police and Cellular Forensic Unit Peshawar-KP, Pakistan. information technologies.Major finding(s): It is evident that the old traditional approach will never provide solutions for better management in controlling crimes. The best way to control crimes and promotion of proactive policing is to adopt new technologies. The study reveals that technology have transformed police more effective and vigilant as compared to traditional policing. The heinous crimes like abduction, missing of an individual, snatching, burglaries and blind murder cases are now traceable with the help of technology.Recommendation(s): From the analysis of the data, it is reflected that Information Technology (IT) expert should be recruited along with research analyst to timely assist and facilitate operational as well as investigation units of police .A mobile locator should be Provided to Cellular Forensic Unit to timely apprehend the criminals .Latest digital analysis software should be provided to equip the Cellular Forensic Unit.

Keywords: crime-prevention, cellular-forensic unit-pakistan, crime prevention-digital-pakistan, crminology-pakistan

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Authors: Achiron Anat, Mathilda Mandel, Mayust Sue, Achiron Reuven, Gurevich Michael

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Introduction: As coronavirus disease 2019 (COVID-19) vaccination is currently spreading around the world, it is of importance to assess the ability of multiple sclerosis (MS) patients to mount an appropriate immune response to the vaccine in the context of disease-modifying treatments (DMT’s). Objectives: Evaluate immunity generated following COVID-19 vaccination in MS patients, and assess factors contributing to protective humoral and cellular immune responses in MS patients vaccinated against severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) virus infection. Methods: Review our recent data related to (1) the safety of PfizerBNT162b2 COVID-19 mRNA vaccine in adult MS patients; (2) the humoral post-vaccination SARS-CoV2 IgG response in MS vaccinees using anti-spike protein-based serology; and (3) the cellular immune response of memory B-cells specific for SARS-CoV-2 receptor-binding domain (RBD) and memory T-cells secreting IFN-g and/or IL-2 in response to SARS-CoV2 peptides using ELISpot/Fluorospot assays in MS patients either untreated or under treatment with fingolimod, cladribine, or ocrelizumab; (4) covariate parameters related to mounting protective immune responses. Results: COVID-19 vaccine proved safe in MS patients, and the adverse event profile was mainly characterised by pain at the injection site, fatigue, and headache. Not any increased risk of relapse activity was noted and the rate of patients with acute relapse was comparable to the relapse rate in non-vaccinated patients during the corresponding follow-up period. A mild increase in the rate of adverse events was noted in younger MS patients, among patients with lower disability, and in patients treated with DMTs. Following COVID-19 vaccination protective humoral immune response was significantly decreased in fingolimod- and ocrelizumab- treated MS patients. SARS-CoV2 specific B-cell and T-cell cellular responses were respectively decreased. Untreated MS patients and patients treated with cladribine demonstrated protective humoral and cellular immune responses, similar to healthy vaccinated subjects. Conclusions: COVID-19 BNT162b2 vaccine proved as safe for MS patients. No increased risk of relapse activity was noted post-vaccination. Although COVID-19 vaccination is new, accumulated data demonstrate differences in immune responses under various DMT’s. This knowledge can help to construct appropriate COVID-19 vaccine guidelines to ensure proper immune responses for MS patients.

Keywords: covid-19, vaccination, multiple sclerosis, IgG

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Authors: Malkhaz Jokhadze, Vakhtang Mshvildadze, Levan Makaradze, Ekaterine Mosidze, Salome Barbaqadze, Mariam Murtazashvili, Dali Berashvili, Koba sivsivadze, Lasha Bakuridze, Aliosha Bakuridze

Abstract:

Essential oils are one of the most important groups of biologically active substances present in plants. Due to the chemical diversity of components, essential oils and their preparations have a wide spectrum of pharmacological action. They have bactericidal, antiviral, fungicidal, antiprotozoal, anti-inflammatory, spasmolytic, sedative and other activities. They are expectorant, spasmolytic, sedative, hypotensive, secretion enhancing, antioxidant remedies. Based on preliminary pharmacological studies, we have developed a formulation called “Phytobiotic” containing essential oils, a feed additive for poultry as an alternative to antibiotics. Phytobiotic is a water-soluble powder containing a composition of essential oils of thyme, clary, monarda and auxiliary substances: dry extract of liquorice and inhalation lactose. On this stage of research, the goal was to study the chemical composition of provided phytobiotic, identify the main substances and determine their quantity, investigate the biological activity of phytobiotic through in vitro and in vivo studies. Using gas chromatography-mass spectrometry, 38 components were identified in phytobiotic, representing acyclic-, monocyclic-, bicyclic-, and sesquiterpenes. Together with identification of main active substances, their quantitative content was determined, including acyclic terpene alcohol β-linalool, acyclic terpene ketone linalyl acetate, monocyclic terpenes: D-limonene and γ-terpinene, monocyclic aromatic terpene thymol. Provided phytobiotic has pronounced and at the same time broad spectrum of antibacterial activity. In the cell model, phytobiotic showed weak antioxidant activity, and it was stronger in the ORAC (chemical model) tests. Meanwhile anti-inflammatory activity was also observed. When fowls were supplied feed enriched with phytobiotic, it was observed that gained weight of the chickens in the experimental group exceeded the same data for the control group during the entire period of the experiment. The survival rate of broilers in the experimental group during the growth period was 98% compared to -94% in the control group. As a result of conducted researches probable four different mechanisms which are important for the action of phytobiotics were identified: sensory, metabolic, antioxidant and antibacterial action. General toxic, possible local irritant and allergenic effects of phytobiotic were also investigated. Performed assays proved that formulation is safe.

Keywords: clary, essential oils, monarda, poultry, phytobiotics, thyme

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Authors: Swapan Maiti, Dipanwita Roy Chowdhury

Abstract:

Nonlinear functions are essential in different cryptoprimitives as they play an important role on the security of the cipher designs. Rule 30 was identified as a powerful nonlinear function for cryptographic applications. However, an attack (MS attack) was mounted against Rule 30 Cellular Automata (CA). Nonlinear rules as well as maximum period CA increase randomness property. In this work, nonlinear rules of maximum period nonlinear hybrid CA (M-NHCA) are studied and it is shown to be a better crypto-primitive than Rule 30 CA. It has also been analysed that the M-NHCA with single nonlinearity injection proposed in the literature is vulnerable against MS attack, whereas M-NHCA with multiple nonlinearity injections provide maximum length cycle as well as better cryptographic primitives and they are also secure against MS attack.

Keywords: cellular automata, maximum period nonlinear CA, Meier and Staffelbach attack, nonlinear functions

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Authors: Surendra Kumar Gautam, Mahesh Dhungana

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Magnesium oxide nanoparticles (MgO NPs) are less toxic to humans and the environment as compared to other metal oxide nanoparticles. Various conventional chemical and physical methods are used for synthesis whose toxicity level is high and highly expensive. As the best alternative, phyto-assisted synthesis has emerged, which uses extracts from plant parts for the synthesis of nanoparticles. Here, we report the synthesis of MgO nanoparticles with the assistance of beetroot extract and leaf extract of P. guajava and A. adenophora. The synthesized MgO NPs were characterized by X-ray diffraction (XRD), Fourier transforms infrared spectroscopy (FTIR), and UV-visible spectroscopy. X-ray analysis for the broadening of peaks was used to evaluate the crystallite size and lattice strain using Debye-Scherer and Williamson–Hall method. The results of crystallite size obtained by both methods are in close proximity. The crystallite size obtained by the Williamson-Hall method seems more accurate, with values being 8.1 nm and 13.2 nm for beetroot MgO NPs and P. guajava MgO NPs, respectively. The FT-IR spectroscopy revealed the dominance of chemical bonds as well as functional groups on MgO NPs surfaces. The UV-visible absorption spectra of MgO NPs were found to be 310 nm, 315 nm, and 315 nm for beetroot, P. guajava, and A. adenophora leaf extract, respectively. Among the three samples, beetroot-mediated MgO NPs were effective antibacterial against both gram-positive and Gram-negative bacteria. In addition, synthesized MgO NPs also show significant antioxidant efficacy against 1,1-diphenyl-2-picrylhydrazyl radical. Further, beetroot MgO NPs showed the highest photocatalytic activity of about 91% in comparison with other samples.

Keywords: MgO NPs, XRD, FTIR, antibacterial, antioxidant and photocatalytic activity

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Authors: Crista Arangala

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Light switching games are both popular and well studied. This paper introduces a cellular automata called Langton’s turmite to several different light switching scenarios and discusses when Langton’s turmite can solve these games.

Keywords: cellular automata, lights out, alien tiles, chaos, Langton's Turmite

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Authors: Ž. Vaštag, Lj. Popović, S. Popović

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After cold pressing of pumpkin oil, the defatted oil cake (PUOC) was utilized as raw material for processing of bio-functional hydrolysates. In this study, the in vitro bioactivity of an alcalase (AH) and a pepsin hydrolysate (PH) prepared from the major pumpkin 12S globulin (cucurbitin) are compared. The hydrolysates were produced at optimum reaction conditions (temperature, pH) for the enzymes, during 60min. The bioactivity testing included antioxidant and angiotensin I converting enzyme inhibitory activity assays. The hydrolysates showed high potential as natural antioxidants and possibly antihypertensive agents in functional food or nutraceuticals. Additionally, preliminary studies have shown that both hydrolysates could exhibit modest α-amylase inhibitory activity, which indicates on their hypoglycemic potential.

Keywords: cucurbitin, alcalase, pepsin, protein hydrolysates, in vitro bioactivity

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Authors: A. K. Asekunowo, A O. T. Asafa, O. O. Okoh, O. T. Asekun, O. B. Familoni

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Background: Acalypha godseffiana is an important plant used both as an ornamental and herbs; its leaves are employed in management of diseases such as diabetics in Eastern Nigeria. Aim: The correlations of the polyphenolic compounds in the hypoglycemic potential of different extracts of leaves of A. godseffiana and their safety profile on cell lines were investigated. Materials and Methods: The phytochemical compositions and antioxidants potentials were determined using adopted methods. An in vitro approach was employed in determining the hypoglycemic potentials of the extracts on α-amylase and α-glucosidase. The Line weaver-Burke plot was used to evaluate the mechanisms of Inhibition mechanisms of the enzymes. Results and Conclusions: Antioxidants results revealed that total antioxidant capacity (TAC) of the acetone extract (IC50: 0.34 mg/mL) showed better activity compared to the standards (silymarine 0.52 mg/mL; gallic acid 0.51 mg/mL). In-vitro hypoglycemic activity of the extracts confirmed that acetone extract demonstrated strong and mild inhibitory potential against α-amylase and α-glucosidase respectively. The observed activity was concentration-dependent with IC50 values of 2.33 and 0.13 mg/mL. The observed hypoglycemic and anti-oxidant potentials of acetone extract A. godseffiana correlate to its high polyphenolic contents which include phenols (133.20 mg gallic acid g-1), flavonoid (350.60 mg quercetin g-1) and tannins (264.67 mg catechin g-1). The mechanisms of action exhibited by acetone extract of A. godseffiana were mixed non-competitive and uncompetitive; which can be attributed to its inhibitory properties on α-amylase and α-glucosidase respectively. This effect would cause reduction in the rate at which starch hydrolyse, boost palliated glucose levels; hence, making acetone extract of A. godseffiana a potential anti-hypoglycemic alternative.

Keywords: Acalypha godeseffiana, acetone extract, anti-hypoglycemia, antioxidant, phytochemicals

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Authors: David Guillermo Piedrahita Marquez, Hector Suarez Mahecha, Jairo Humberto Lopez

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The propolis are substances obtained from the beehive have an action against pathogens, prooxidant substances and free radicals because of its polyphenols content, this has motivated the use of these compounds in the food and pharmaceutical industries. However, due to their organoleptic properties and their ability to react with other compounds, their application has been limited; therefore, the objective of this research was to propose a mechanism to protect propolis and mitigate side effects granted by its components. To achieve the stated purpose ethanolic extracts of propolis (EEP) from three samples from Santander were obtained and their antioxidant and antimicrobial activity were evaluated in order to choose the extract with the biggest potential. Subsequently mixtures of the extract with maltodextrin were prepared by spray drying varying concentration and temperature, finally the yield, the physicochemical, and antioxidant properties of the products were measured. It was concluded that Socorro propolis was the best for the production of microencapsulated due to their activity against pathogenic strains, for its large percentage of DPPH radical inactivation and for its high phenolic content. In spray drying, the concentration of bioactive had a greater impact than temperature and the conditions set allowed a good performance and the production of particles with high antioxidant potential and little chance of proliferation of microorganisms. Also, it was concluded that the best conditions that allowed us to obtain the best particles were obtained after drying a mixture 1:2 ( EEP: Maltodextrin), besides the concentration is the most important variable in the spray drying process, at the end we obtained particles of different sizes and shape and the uniformity of the surface depend on the temperature. After watching the previously mentioned microparticles by scanning electron microscopy (SEM) it was concluded that most of the particles produced during the spray dry process had a spherical shape and presented agglomerations due to the moisture content of the ethanolic extracts of propolis (EEP), the morphology of the microparticles contributed to the stability of the final product and reduce the loss of total phenolic content.

Keywords: spray drying, propolis, maltodextrin, encapsulation, scanning electron microscopy

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Authors: Djebbar Atmani, Aghiles Karim Aissat, Nadjet Debbache-Benaida, Nassima Chaher-Bazizi, Dina Atmani-Kilani, Meriem Rahmani-Berboucha, Naima Saidene, Malika Benloukil, Lila Azib

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Medicinal plants are believed to be an important source for the discovery of potential antioxidant, anti-inflammatory and anti-diabetic substances. The present study was designed to investigate the neuroprotective, anti-inflammatory, anti-diabetic and anti-hyperuricemic potential of Pistacia lentiscus, as well as the identification of active compounds. The antioxidant potential of plant extracts against known radicals was measured using various standard in vitro methods. Anti-inflammatory activity was determined using the paw edema model in mice and by measuring the secretion of the pro-inflammatory cytokine, whereas the anti-diabetic effect was assessed in vivo on streptozotocin-induced diabetic rats and in vitro by inhibition of alpha-amylase. The anti-hyperuricemic activity was evaluated using the xanthine oxidase assay, whereas neuroprotective activity was investigated using an Aluminum-induced toxicity test. Pistacia lentiscus extracts and fractions exhibited high scavenging capacity against DPPH, NO. and ABTS+ radicals in a dose-dependent manner and restored blood glucose levels, in vivo, to normal values, in agreement with the in vitro anti-diabetic effect. Oral administration of plant extracts significantly decreased carrageenan-induced mice paw oedema, similar to the standard drug, diclofenac, was effective in reducing IL-1β levels in cell culture and induced a significant increase in urinary volume in mice, associated to a promising anti-hyperuricemic activity. Plant extracts showed good neuroprotection and restoration of cognitive functions in mice. HPLC-MS and NMR analyses allowed the identification of known and new phenolic compounds that could be responsible for the observed activities. Therefore, Pistacia lentiscus could be beneficial in the treatment of inflammatory conditions and diabetes complications and the enhancement of cognitive functions.

Keywords: Pistacia lentiscus, anti-inflammatory, antidiabetic, flavanols, neuroprotective

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Authors: Fatemeh Haghiralsadat, Mohadese Hashemi, Elham Akhoundi Kharanaghi, Mojgan Yazdani, Mahboobe Sharafodini, Omid Javani

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Niosomes are colloidal particles formed from the self-assembly of non-ionic surfactants in aqueous medium resulting in closed bilayer structures. As a consequence of this hydrophilic and hydrophobic structure, niosomes have the capacity to entrap compounds of different solubilities. Niosomes are promising vehicle for drug delivery which protect sensitive drugs and improve the therapeutic index of drugs by restricting their action to target cells. Essential oils are complex mixtures of volatile compounds such as terpenoids, phenol-derived aromatic components that have been used for many biological properties including bactericidal, fungicidal, insecticidal, antioxidant, anti-tyrosinase and other medicinal properties. Encapsulation of essential oils in niosomes can be an attractive method to overcome their limitation such as volatility, easily decomposition by heat, humidity, light, or oxygen. Cuminum cyminum L. (Cumin) is an aromatic plant included in the Apiaceae family and is used to flavor foods, added to fragrances, and for medical preparations which is indigenous to Egypt, the Mediterranean region, Iran and India. The major components of the Cumin oil were reported as cuminaldehyde, γ -terpinene, β-pinene, p-cymene, p-mentha-1, 3-dien-7-al, and p-mentha-1, 4-dien-7-al which provide the antimicrobial and antioxidant activity. The aim of this work was to formulate Cumin essential oil-loaded niosomes to improve water solubility of natural product and evaluate its physico-chemical features and stability. Cumin oil was obtained through steam distillation using a clevenger-type apparatus and GC/MS was applied to identify the main components of the essential oil. Niosomes were prepared by using thin film hydration method and nanoparticles were characterized for particle size, dispersity index, zeta potential, encapsulation efficiency, in vitro release, and morphology.

Keywords: Cuminum cyminum L., Cumin, niosome, essential oil, encapsulation

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Authors: Cristian Moisa, Andreea Lupitu, Adriana Csakvari, Dana G. Radu, Leonard Marian Olariu, Georgeta Pop, Dorina Chambre, Lucian Copolovici, Dana Copolovici

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Green synthesis of nanoparticles (NPs) represents a promising, accessible, eco-friendly, and safe process with significant applications in biotechnology, pharmaceutical sciences, and farming. The aim of our study was to obtain silver nanoparticles, using plant wastes extracts resulted in the essential oils extraction process: Thymus vulgaris L., Origanum vulgare L., Lavandula angustifolia L., and in hemp processing for seed and fibre, Cannabis sativa. Firstly, we obtained aqueous extracts of thyme, oregano, lavender, and hemp (two monoicous and one dioicous varieties), all harvested in western part of Romania. Then, we determined the chemical composition of the extracts by liquid-chromatography coupled with diode array and mass spectrometer detectors. The compounds identified in the extracts were in agreement with earlier published data, and the determination of the antioxidant activity of the obtained extracts by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assays confirmed their antioxidant activity due to their total polyphenolic content evaluated by Folin-Ciocalteu assay. Then, the silver nanoparticles (AgNPs) were successfully biosynthesised, as was demonstrated by UV-VIS, FT-IR spectroscopies, and SEM, by reacting AgNO₃ solution and plant extracts. AgNPs were spherical in shape, with less than 30 nm in diameter, and had a good bactericidal activity against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas fluorescens).

Keywords: plant wastes extracts, chemical composition, high performance liquid chromatography mass spectrometer, HPLC-MS, scanning electron microscopy, SEM, silver nanoparticles

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Authors: Farhana Nosheen, Moeez Ahmad

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Throughout the food chain, the food waste from initial agricultural production to final household consumption has become a serious concern for global sustainability because of its adverse impacts on food security, natural resources, the environment, and human health. About a third of tomatoes (Lycopersicon esculentum L.) delivered to processing plants end as processing waste. The amount of such waste material is estimated to have increased with the emergence of mechanical harvesting. Experiments were made to determine the nutritional profile and antioxidant activity of tomato processing waste and to explore the bioactive compound in tomato waste, i.e., Lycopene. Tomato Variety of ‘SAHARA F1’ was used to make tomato waste. The tomatoes were properly cleaned, and then unwanted impurities were removed properly. The tomatoes were blanched at 90 ℃ for 5 minutes. After which, the skin of the tomatoes was removed, and the remaining part passed through the electric pulper. The pulp and seeds were collected separately. The seeds and skin of tomatoes were mixed and saved in a sterilized jar. The samples of tomato waste were found to contain 89.11±0.006 g/100g moisture, 10.13±0.115 g/100g protein, 2.066±0.57 g/100g fat, 4.81±0.10 g/100g crude fiber, and 4.06±0.057 g/100g ash and NFE 78.92±0.066 g/100g. The results confirmed that tomato waste contains a considerable amount of Lycopene 51.0667±0.00577 mg/100g and exhibited good antioxidant properties. Total phenolics showed average contents of 122.9600±0.01000 mg GAE/100g, of which flavonoids accounted for 41.5367±0.00577 mg QE/100g. Antioxidant activity of tomato processing waste was found 0.6833±0.00577 mmol Trolox/100g. Unsaturated fatty acids represent the major portion of total fatty acids, Linoleic acid being the major one. The mineral content of tomato waste showed a good amount of potassium 3030.1767 mg/100g and calcium 131.80 mg/100g, respectively were present in it. These findings suggest that tomato processing waste is rich in nutrients, antioxidants, fatty acids, and minerals. I recommend that this waste should be sun-dried to be used in the combination of feed of the animals. It can also be used in making some other products like lycopene tea or several other health-beneficial products.

Keywords: food waste, tomato, bioactive compound, sustainable management

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Authors: Kenia Martínez, Geniel Talavera, Juan Alonso

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Moringa oleifera is a plant containing many nutrients that are mostly concentrated within the leaves. Commonly, the separation process of these nutrients involves solid-liquid extraction followed by evaporation and drying to obtain a concentrated extract, which is rich in proteins, vitamins, carbohydrates, and other essential nutrients that can be used in the food industry. In this work, three drying methods were used, which involved very different temperature and pressure conditions, to evaluate the effect of each method on the vitamin C content and the antioxidant efficiency of the extracts. Solid-liquid extractions of Moringa leaf (LE) were carried out by employing an ethanol solution (35% v/v) at 50 °C for 2 hours. The resulting extracts were then dried i) in a convective oven (CO) at 100 °C and at an atmospheric pressure of 750 mbar for 8 hours, ii) in a vacuum evaporator (VE) at 50 °C and at 300 mbar for 2 hours, and iii) in a freeze-drier (FD) at -40 °C and at 0.050 mbar for 36 hours. The antioxidant capacity (EC₅₀, mg solids/g DPPH) of the dry solids was calculated by the free radical inhibition method employing DPPH˙ at 517 nm, resulting in a value of 2902.5 ± 14.8 for LE, 3433.1 ± 85.2 for FD, 3980.1 ± 37.2 for VE, and 8123.5 ± 263.3 for CO. The calculated antioxidant efficiency (AE, g DPPH/(mg solids·min)) was 2.920 × 10^-5 for LE, 2.884 × 10^-5 for FD, 2.512 × 10^-5 for VE, and 1.009 × 10^-5 for CO. Further, the content of vitamin C (mg/L) determined by HPLC was 59.0 ± 0.3 for LE, 49.7 ± 0.6 for FD, 45.0 ± 0.4 for VE, and 23.6 ± 0.7 for CO. The results indicate that the convective drying preserves vitamin C and antioxidant efficiency to 40% and 34% of the initial value, respectively, while vacuum drying to 76% and 86%, and freeze-drying to 84% and 98%, respectively.

Keywords: antioxidant efficiency, convective drying, freeze-drying, Moringa oleifera, vacuum drying, vitamin C content

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Authors: Orsolya Horvath, Laszlo Deres, Krisztian Eros, Katalin Ordog, Tamas Habon, Balazs Sumegi, Kalman Toth, Robert Halmosi

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Introduction: Oxidative stress induces an imbalance in mitochondrial fusion and fission processes, finally leading to cell death. The two antioxidant molecules, BGP-15 and L2286 have beneficial effects on mitochondrial functions and on cellular oxidative stress response. In this work, we studied the effects of these compounds on the processes of mitochondrial quality control. Methods: We used H9c2 cardiomyoblast and isolated neonatal rat cardiomyocytes (NRCM) for the experiments. The concentration of stressors and antioxidants was beforehand determined with MTT test. We applied 1-Methyl-3-nitro-1-nitrosoguanidine (MNNG) in 125 µM, 400 µM and 800 µM concentrations for 4 and 8 hours on H9c2 cells. H₂O₂ was applied in 150 µM and 300 µM concentration for 0.5 and 4 hours on both models. L2286 was administered in 10 µM, while BGP-15 in 50 µM doses. Cellular levels of the key proteins playing role in mitochondrial dynamics were measured in Western blot samples. For the analysis of mitochondrial network dynamics, we applied electron microscopy and immunocytochemistry. Results: Due to MNNG treatment the level of fusion proteins (OPA1, MFN2) decreased, while the level of fission protein DRP1 elevated markedly. The levels of fusion proteins OPA1 and MNF2 increased in the L2286 and BGP-15 treated groups. During the 8 hour treatment period, the level of DRP1 also increased in the treated cells (p < 0.05). In the H₂O₂ stressed cells, administration of L2286 increased the level of OPA1 in both H9c2 and NRCM models. MFN2 levels in isolated neonatal rat cardiomyocytes raised considerably due to BGP-15 treatment (p < 0.05). L2286 administration decreased the DRP1 level in H9c2 cells (p < 0.05). We observed that the H₂O₂-induced mitochondrial fragmentation could be decreased by L2286 treatment. Conclusion: Our results indicated that the PARP-inhibitor L2286 has beneficial effect on mitochondrial dynamics during oxidative stress scenario, and also in the case of directly induced DNA damage. We could make the similar conclusions in case of BGP-15 administration, which, via reducing ROS accumulation, propagates fusion processes, this way aids preserving cellular viability. Funding: GINOP-2.3.2-15-2016-00049; GINOP-2.3.2-15-2016-00048; GINOP-2.3.3-15-2016-00025; EFOP-3.6.1-16-2016-00004; ÚNKP-17-4-I-PTE-209

Keywords: H9c2, mitochondrial dynamics, neonatal rat cardiomyocytes, oxidative stress

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Authors: Sandeep Goyal, Sandeep Saluja

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Natural antioxidants have major role in maintenance of health. Green tea extract principally contains epigallocatechin-3-gallate (EGCG), as its abundant antioxidant constituent. Green tea is consumed daily worldwide as antioxidant to combat CNS diseases and has traditional importance also. EGCG has neuroprotective potential in various animal models of Parkinson disease, Alzheimer’s disease etc. but its exact mechanism has not been ruled out. The present study has been designed to investigate the anti-inflammatory, antioxidant and mitochondrial modulating mechanism of neuroprotective effect of epigallocatechin-3-gallate against rodent model of rotenone induced Parkinson’s disease (PD). The behavioural alterations were assessed by using open field test apparatus, Chatilon’s grip strength test apparatus and elevated plus maze for determining the locomotor activity, grip strength and cognition respectively. Biochemically, various parameters to assess oxidative stress, neuroinflammation and neurochemical estimations were performed on rat brain homogenates. A histological examination of rat brain striatum was done to check the neurodegeneration. Epigallocatechin-3-gallate (EGCG) at 10 & 20 mg/kg, were investigated for their neuroprotective potential along with levodopa as a standard agent. Minocycline, a microglial activation inhibitor, was administered alone and in combination with EGCG. EGCG and minocycline produced ameliorative effect against rotenone induced PD like symptoms by significantly reduced behavioral, biochemical and histological alterations. Results of our study reveal the neuroprotective effect of EGCG and minocycline against rotenone induced PD. Results of our study indicate that EGCG exerted neuroprotective effect against rotenone induced PD via its antioxidant, anti-inflammatory and mitochondrial modulating mechanisms and substantiate its previously reported and traditional claims for its use in CNS diseases.

Keywords: antioxidants, neurotoxicity, rotenone, EGCG

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Authors: Ayse Demirbas, Bruce A. Welt, Yavuz Yagiz

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Oxidation of polyunsaturated fatty acids (PUFA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in fish causes loss of product quality. Oxidative rancidity causes loss of nutritional value and undesirable color changes. Therefore, powerful antioxidant extracts may provide a relatively low cost and natural means to reduce oxidation, resulting in longer, higher quality and higher value shelf life of foods. In this study, we measured effects of red cabbage antioxidant on lipid oxidation in fresh tilapia filets using thiobarbituric acid reactive substances (TBARS) assay, peroxide value (PV) and color assesment analysis. Extraction of red cabbage was performed using an efficient microwave method. Fresh tilapia filets were dipped in or sprayed with solutions containing different concentrations of extract. Samples were stored for up to 9 days at 4°C and analyzed every other day for color and lipid oxidation. Results showed that treated samples had lower oxidation than controls. Lipid peroxide values on treated samples showed benefits through day-7. Only slight differences were observed between spraying and dipping methods. This work shows that red cabbage antioxidant extracts may represent an inexpensive and all natural method for reducing oxidative spoilage of fresh fish.

Keywords: antioxidant, shelf life, fish, red cabbage, lipid oxidation

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