Search results for: DPPH scavenging assay

Authors: Abhijit Das, Runu Chakraborty

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Tea is the most widely consumed beverage aside from water; it is grown in about 30 countries with a per capita worldwide consumption of approximately 0.12 liter per year. Green tea is of growing importance with its antioxidant contents associated with its health benefits. The various extraction methods can influence the polyphenol concentrations of green tea. The purpose of the study was to quantify the polyphenols, flavonoid and antioxidant activity of both caffeinated and decaffeinated form of tea manufactured commercially in Assam, North Eastern part of India. The results display that phenolic/flavonoid content well correlated with antioxidant activity which was performed by DPPH (2,2-diphenyl-1-picrylhydrazyl) and FRAP (Ferric reducing ability of plasma) assay. After decaffeination there is a decrease in the polyphenols concentration which also affects the antioxidant activity of green tea. For the enrichment of antioxidant activity of decaffeinated tea a herbal plant extract is used which shows a synergistic effect between green tea and herbal plant phenolic compounds.

Keywords: antioxidant activity, decaffeination, green tea, flavonoid content, phenolic content, plant extract

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Authors: Mi-Ju Kim, Hae-Yeong Kim

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Product mislabeling and adulteration have been increasing the concerns in processed meat products. Relatively inexpensive pork meat compared to meat such as beef was adulterated for economic benefit. These food fraud incidents related to pork were concerned due to economic, religious and health reasons. In this study, a rapid on-site detection method using loop-mediated isothermal amplification (LAMP) was developed for the simultaneous identification of beef and pork. Each specific LAMP primer for beef and pork was designed targeting on mitochondrial D-loop region. The LAMP assay reaction was performed at 65 ℃ for 40 min. The specificity of each primer for beef and pork was evaluated using DNAs extracted from 13 animal species including beef and pork. The sensitivity of duplex LAMP assay was examined by serial dilution of beef and pork DNAs, and reference binary mixtures. This assay was applied to processed meat products including beef and pork meat for monitoring. Each set of primers amplified only the targeted species with no cross-reactivity with animal species. The limit of detection of duplex real-time LAMP was 1 pg for each DNA of beef and pork and 1% pork in a beef-meat mixture. Commercial meat products that declared the presence of beef and/or pork meat on the label showed positive results for those species. This method was successfully applied to detect simultaneous beef and pork meats in processed meat products. The optimized duplex LAMP assay can identify simultaneously beef and pork meat within less than 40 min. A portable real-time fluorescence device used in this study is applicable for on-site detection of beef and pork in processed meat products. Thus, this developed assay was considered to be an efficient tool for monitoring meat products.

Keywords: beef, duplex real-time LAMP, meat identification, pork

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Authors: Kar Lin Nyam, Phey Yee Lim

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The aim of this study was to determine the antioxidant potential, nutritional composition, and functional properties of kenaf leaves powder. Besides, the effect of kenaf leaves powder in bread qualities, properties, and consumer acceptability were evaluated. Different formulations of bread fortified with 0%, 4% and 8% kenaf leaves powder, respectively were produced. Physical properties of bread, such as loaf volume, dough expansion, crumb colour, and bread texture, were determined. Nine points hedonic scale was utilized in sensory evaluation to determine the best formulation (the highest overall acceptability). Proximate composition, calcium content, and antioxidant properties were also determined for the best formulation. 4% leaves powder bread was the most preferred by the panelists followed by control bread, and the least preferred was being 8% leaves powder bread. 4% leaves powder bread had significantly higher value of DPPH radical scavenging capacity (8.05 mg TE/100g), total phenolic content (12.88 mg GAE/100g) and total flavonoid content (13.26 mg QE/100g) compared to control bread (1.38 mg TE/100g, 8.17 mg GAE/100g, and 8.77 mg QE/100g respectively). Besides, 4% leaves powder bread also showed higher in calcium content and total dietary fiber compared to control bread. Kenaf leaves powder is suitable to be used as a source of natural antioxidant for fortification and nutrient improver in bread.

Keywords: dietary fibre, calcium, total phenolic content, total flavonoid content

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Authors: Liga Priecina, Daina Karklina

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Antioxidants are became the most analyzed substances in last decades. Antioxidants act as in activator for free radicals. Spices and vegetables are one of major antioxidant sources. Most common antioxidants in vegetables and spices are vitamin C, E, phenolic compounds, carotenoids. Therefore, it is important to get some view about antioxidant changes in spices and vegetables during processing. In this article was analyzed nine fresh and dried spices and vegetables- celery (Apium graveolens), parsley (Petroselinum crispum), dill (Anethum graveolens), leek (Allium ampeloprasum L.), garlic (Allium sativum L.), onion (Allium cepa), celery root (Apium graveolens var. rapaceum), pumpkin (Curcubica maxima), carrot (Daucus carota)- grown in Latvia 2013. Total carotenoids and phenolic compounds and their antiradical scavenging activity were determined for all samples. Dry matter content was calculated from moisture content. After drying process carotenoid content significantly decreases in all analyzed samples, except one -carotenoid content increases in parsley. Phenolic composition was different and depends on sample – fresh or dried. Total phenolic, flavonoid and phenolic acid content increases in dried spices. Flavan-3-ol content is not detected in fresh spice samples. For dried vegetables- phenolic acid content decreases significantly, but increases flavan-3-ols content. The higher antiradical scavenging activity was observed in samples with higher flavonoid and phenolic acid content.

Keywords: antiradical scavenging activity, carotenoids, phenolic compounds, spices, vegetables

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Authors: Mohamed H. El-Newehy, Sameh M. Osman, Moamen S. Refat, Salem S. Al-Deyab, Ayman El-Faham

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The two copolymers itaconic acid-methyl methacrylate and itaconic acid-acrylamide have been prepared in different ratio by radical copolymerization in the presence of azobisisobutyronitrile (AIBN) as initiator and using 2-butanone as reaction medium using microwave irradiation. The microwave technique is safe, fast, and gives high yield of the products with high purity in an optimum time, comparing to the traditional conventional heating. All the prepared copolymers were characterized by FT-IR, thermal analysis and elemental microanalysis. The itaconic acid-based copolymers showed a good sensitivity in alkaline media for scavenging Cu (II) and Pb (II). The chelation behavior of both Cu (II) and Pb (II) complexes were checked using FT-IR, thermogravimetric analysis (TGA), and differential scanning calorimetery (DSC). The infrared data are in a good agreement with the coordination through carboxylate-to-metal, in which the copolymers acting as a bidentate ligand.

Keywords: microwave synthesis, itaconic acid, copolymerization, scavenging, thermal stability

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Authors: Supriyono Supriyono, Sumardiyono Sumardiyono, Peni Pujiastuti, Dian Indriana Hapsari

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Crude palm oil (CPO) is a vegetable oil that came from a palm tree bunch. The productivity of the oil is 12 ton/hectare/year. Thus palm oil tree was known as highest vegetable oil yield. It was grown across Equatorial County, especially in Malaysia and Indonesia. The greenish-red color on CPO was come from carotenoid. Carotenoid is one of the antioxidants that could be extracted. Carotenoid could be used as functional food and other purposes. Another antioxidant that also found in CPO is tocopherol. The aim of the research work is to find antioxidant activity on CPO comparing to the synthetic antioxidant that available in a market. In this research work, antioxidant was extracted by a mixture of acetone and n.hexane, while the activity of the antioxidant extract was determined by DPPH method. Antioxidant activity of the extracted compound about 46% compared to pure tocopherol. While the solvent mixture compose by 90% acetone and 10% n. hexane meet the best on the antioxidant activity.

Keywords: antioxidant, beta carotene, crude palm oil, DPPH, tocopherol

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Authors: Tunca Gul Altuntas, Aziz Baydar, Cemre Acar, Sezen Yılmaz, Tulay Coban

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Free radicals, which are generated in many bioorganic redox processes, play a role in the pathogenesis of several diseases including cancer, arthritis, hemorrhagic shock, inflammatory, cardiovascular, neurodegenerative diseases and age-related degenerative brain diseases. Exposures of normal cell to free radical damages several structures, oxidizes nucleic acids, proteins, lipids, or DNA. Compounds interfere with the action of reactive oxygen species might be useful in prevention and treatment of these pathologies. A series of indole compounds containing piperazine ring were synthesized. Coupling of indole-2-carboxylic acid with monosubstituted piperazines was accomplished with 1,1’-carbonyldiimidazole (CDI) in a good yield. The structures of prepared compounds were verified in good agreement with their 1H NMR (nuclear magnetic resonance), MS (mass spectrophotometry), and IR (infrared spectrophotometry) characteristics. In this work, all synthetized indole derivatives were screened in vitro for their antioxidative potential against vitamin E (α-tocopherol) using different antioxidant assays such as superoxide anion formation, lipid peroxidation levels in rat liver, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) stable radical scavenging activity. The synthesized compounds showed various levels of inhibition compared to vitamin E. This may give promising results for the development of new antioxidant agents.

Keywords: antioxidant, indoles, piperazines, reactive oxygen species

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Authors: Samar Saadeldin Abdelmotalab Omer, Ikram Mohammed Eltayeb Elsiddig, Amna Beshir Medani Ahmed, Saad Mohammed Hussein Ayoub

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Many herbal medicinal products are considered as potential hypocholesterolemic agents with encouraging safety profiles, however, only a limited amount of clinical research exists to support their efficacy. The present study was designed to compare the antihypercholesterolaemic and antioxidant activities of the crude ethanolic extracts of Citrus reticulata peel, Zingiber officinale rhizome, and Sesamum indicum seeds. Forty-five rats were used throughout the experiment, which were divided into nine groups, five rats in each as follows; normal control group (normal rats fed with standard normal diet), rats fed hypercholesterolemic diet consisting of 1% cholesterol and 10% saturated animal fat, which were further divided into eight groups; hypercholesterolemic control group (rats only fed hypercholesterolemic diet), groups 3,4,5,6,7, and 8 were given Citrus reticulata, Zingiber officinale, and Sesamum indicum ethanolic extracts at doses of (250mg/kg and 500mg/kg, respectively) orally; and group 9 rats were given atorvastatin (0.18mg/kg) orally as a reference antihypercholesterolaemic drug. Blood samples were obtained four weeks following treatment from the retro-orbital venous plexus after fasting overnight from all groups and the lipid profile (serum total cholesterol (TC), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and triglycerides levels) was measured and the risk ratio (TC/HDL-C) was assessed. The antioxidant activity of the three plant extracts was determined using DPPH free-radical assay. Results of in vivo and in vitro antihypercholesterolaemic and antioxidant assay, respectively, revealed that the three extracts possess comparable antioxidant and anti-hypercholesterolaemic activities.

Keywords: anti hypercholesterolemic effects, antioxidant activity, HDL, LDL, TC, TGs, citrus reticulata, sesamum indicum, zingiber officinale

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Authors: Farah El Mohtadi, Richard d'Arcy, Nicola Tirelli

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Since 40 years, poly (ethylene glycol) (PEG) has been the gold standard in biomaterials and drug delivery, because of its combination of chemical and biological inertness. However, the possibility of its breakdown under oxidative conditions and the demonstrated development of anti-PEG antibodies highlight the necessity to develop carriers based on materials with increased stability in a challenging biological environment. Here, we describe the synthesis of polysulfide via anionic ring-opening polymerization. In vitro, the synthesized polymer was characterized by low toxicity and a level of complement activation (in human plasma) and macrophage uptake slightly lower than PEG and poly (2‐methyl-2‐oxazoline) (PMOX), of a similar size. Importantly, and differently from PEG, on activated macrophages, the synthesized polymer showed a strong and dose-dependent ROS scavenging activity, which resulted in the corresponding reduction of cytokine production. Therefore, the results from these studies show that polysulfide is highly biocompatible and are potential candidates to be used as an alternative to PEG for various applications in nanomedicine.

Keywords: PEG, low toxicity, ROS scavenging, biocompatible

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Authors: Thilivhali Emmanuel Tshikalange, Phiwokuhle Mamba

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Ten medicinal plants used traditionally in the treatment of sexually transmitted diseases (STDs) and urinary tract infections (UTIs) were selected from an ethnobotanical database developed in Mpumalanga. The plants were investigated for their antimicrobial activity against five bacterial strains (Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Neisseria gonorrhoeae and Staphylococcus aureus) and one fungal strain (Candida albicans). Eight of the plants inhibited the growth of all microorganisms at a concentration range of 0.4 mg/ml to 12.5 mg/ml. Acacia karroo showed the most promising antimicrobial activity, with a minimum inhibitory concentration (MIC) of 0.4 mg/ml on Staphylococcus aureus and 0.8 mg/ml on Neisseria gonorrhoeae. All ten plants were further investigated for their antioxidant activities using the DPPH scavenging method. Acacia karroo and Rhoicissus tridentata subsp. cuneifolia showed good antioxidant activity with IC50 values of 0.83 mg/ml and 0.06 mg/ml, respectively. The toxicity of plants was determined using the XTT reduction method against Vero cells. None of the ten plants showed toxicity on the cells. The obtained results confirmed that Acacia karroo and possibly Rhoicissus tridentata subsp. cuneifolia have the potential of being used as antimicrobial agents in the treatment of STDs and UTIs. These results support and validate traditional use of medicinal plants studied.

Keywords: antimicrobial, antioxidant, Neisseria gonorrhoeae, sexually transmitted diseases

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Authors: Maria P. Geneva, Ira V. Stancheva, Marieta G. Hristozkova, Roumiana D. Vasilevska-Ivanova, Mariana T. Sichanova, Janet R. Mincheva

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Hyssopus officinalis L., Lamiaceae, commonly called hyssop, is an aromatic, semi-evergreen, woody-based, shrubby perennial plant. Hyssop is a good expectorant and antiviral herb commonly used to treat respiratory conditions such as influenza, sinus infections, colds, and bronchitis. Most of its medicinal properties are attributed to the essential oil of hyssop. The study was conducted to evaluate the influence of inoculation with arbuscular mycorrhizal fungi of in vitro propagated hyssop plants on the: activities of antioxidant enzymes superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase; accumulation of non-enzymatic antioxidants total phenols and flavonoid, water-soluble soluble antioxidant metabolites expressed as ascorbic acid; the antioxidant potential of hyssop methanol extracts assessed by two common methods: free radical scavenging activity using free stable radical (2,2-diphenyl-1-picrylhydrazyl, DPPH• and ferric reducing antioxidant power FRAP in flowers and leaves. The successfully adapted to field conditions in vitro plants (survival rate 95%) were inoculated with arbuscular mycorrhizal fungi (Claroideoglomus claroideum, ref. EEZ 54). It was established that the activities of enzymes with antioxidant capacity (superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase) were significantly higher in leaves than in flowers in both control and mycorrhized plants. In flowers and leaves of inoculated plants, the antioxidant enzymes activity were lower than in non-inoculated plants, only in SOD activity, there was no difference. The content of low molecular metabolites with antioxidant capacity as total phenols, total flavonoids, and water soluble antioxidants was higher in inoculated plants. There were no significant differences between control and inoculated plants both for FRAP and DPPH antioxidant activity. According to plant essential oil content, there was no difference between non-inoculated and inoculated plants. Based on our results we could suggest that antioxidant capacity of in vitro propagated hyssop plant under conditions of cultivation is determined by the phenolic compounds-total phenols and flavonoids as well as by the levels of water-soluble metabolites with antioxidant potential. Acknowledgments: This study was conducted with financial support from National Science Fund at the Bulgarian Ministry of Education and Science, Project DN06/7 17.12.16.

Keywords: antioxidant enzymes, antioxidant metabolites, arbuscular mycorrhizal fungi, Hyssopus officinalis L.

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Authors: Mohamed M. Ibrahima, Abdul Aziz A.AL Sahli, Ibrahim A. Alaraidh, Ali A. Al-Homaidan, E.M. Mostafa, G.A. EL-Gaaly

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Traditionally, in Middle Eastern countries, many cultures use chewing sticks of arak for medicinal purposes especially, for oral cleanliness care. It was used by Muslims for the treatment of teeth and highly recommended to be used by muslims during the whole day. Therefore, the present work aimed to determine the total phenolic content and total flavonoids in two Miswak extracts obtained from arak roots collected from two different localities in Saudi Arabia. They were extracted with aqueous ethanol (80%) and used to estimate in vitro their antioxidative abilities. The new findings showed that the two tested extracts contained significantly different amounts of both total phenolic content and total flavonoids. According to the increase of total phenolic contents and total flavonoids obtained from the two extracts, Miswak collected from the southern region was found to contain more contents than those collected from the middle region. The results of antioxidant activities of Miswak root extract obtained by using different in vitro methods were varied depending on the technique used. According to the malondialdehyde (MDA) method, hydrogen peroxide (H2O2) scavenging ability and 1,1-diphenyl-2-picrylhydrazyl (DPPH) methods, the two Miswak extracts exhibited to have high to very high antioxidant activities. Mostly, the values of antioxidant activities of Southern region have been shown to be always the highest.

Keywords: Arak, antioxidant, medicinal plants, Saudi Arabia

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Authors: Moon Jung Kim, Guil Rhim

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The clinical sensitivity of the “VEUPLEXTM TBI assay”, a clinical trial medical device, in mild traumatic brain injury was 28.6% (95% CI, 19.7%-37.5%), and the clinical specificity was 94.0% (95% CI, 89.3%). -98.7%). In addition, when the results analyzed by marker were put together, the sensitivity was higher when interpreting the two tests together than the two tests, UCHL1 and GFAP alone. Additionally, when sensitivity and specificity were analyzed based on CT results for the mild traumatic brain injury patient group, the clinical sensitivity for 2 CT-positive cases was 50.0% (95% CI: 1.3%-98.7%), and 19 CT-negative cases. The clinical specificity for cases was 68.4% (95% CI: 43.5% - 87.4%). Since the low clinical sensitivity for the two CT-positive cases was not statistically significant due to the small number of samples analyzed, it was judged necessary to secure and analyze more samples in the future. Regarding the clinical specificity analysis results for 19 CT-negative cases, there were a large number of patients who were actually clinically diagnosed with mild traumatic brain injury but actually received a CT-negative result, and about 31.6% of them showed abnormal results on VEUPLEXTM TBI assay. Although traumatic brain injury could not be detected in 31.6% of the CT scans, the possibility of actually suffering a mild brain injury could not be ruled out, so it was judged that this could be confirmed through follow-up observation of the patient. In addition, among patients with mild traumatic brain injury, CT examinations were not performed in many cases because the symptoms were very mild, but among these patients, about 25% or more showed abnormal results in the VEUPLEXTM TBI assay. In fact, no damage is observed with the naked eye immediately after traumatic brain injury, and traumatic brain injury is not observed even on CT. But in some cases, brain hemorrhage may occur (delayed cerebral hemorrhage) after a certain period of time, so the patients who did show abnormal results on VEUPLEXTM TBI assay should be followed up for the delayed cerebral hemorrhage. In conclusion, it was judged that it was difficult to judge mild traumatic brain injury with the VEUPLEXTM TBI assay only through clinical findings without CT results, that is, based on the GCS value. Even in the case of CT, it does not detect all mild traumatic brain injury, so it is difficult to necessarily judge that there is no traumatic brain injury, even if there is no evidence of traumatic brain injury in CT. And in the long term, more patients should be included to evaluate the usefulness of the VEUPLEXTM TBI assay in the detection of microscopic traumatic brain injuries without using CT.

Keywords: brain injury, traumatic brain injury, GFAP, UCHL1

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Authors: Eman Abdullah Morsi, Hend Okasha, Heba Abdel Hady, Mortada El-Sayed, Mohamed Abbas Shemis

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Context: Linum usitatissimum (Linn), known as Flaxseed, is one of the most important medicinal plants traditionally used for various health as nutritional purposes. Objective: Estimation of total phenolic and flavonoid contents as well as evaluate the antioxidant using α, α-diphenyl-β-picrylhydrazyl (DPPH), 2-2'azinobis (3-ethylbenzthiazoline-6-sulphonic acid (ABTS) and total antioxidant capacity (TAC) assay and investigation of anti-inflammatory by Bovine serum albumin (BSA) and anticancer activities of hepatocellular carcinoma cell line (HepG2) and breast cancer cell line (MCF7) have been applied on hexane, ethyl acetate, n-butanol and methanol extracts and also, fractions of methonal extract (hexane, ethyl acetate and n-butanol). Materials and Methods: Phenolic and flavonoid contents were detected using spectrophotometric and colorimetric assays. Antioxidant and anti-inflammatory activities were estimated in-vitro. Anticancer activity of extracts and fractions of methanolic extract were tested on (HepG2) and (MCF7). Results: Methanolic extract and its ethyl acetate fraction contain higher contents of total phenols and flavonoids. In addition, methanolic extract had higher antioxidant activity. Butanolic and ethyl acetate fractions yielded higher percent of inhibition of protein denaturation. Meanwhile, ethyl acetate fraction and methanolic extract had anticancer activity against HepG2 and MCF7 (IC50=60 ± 0.24 and 29.4 ± 0.12µg.mL⁻¹) and (IC50=94.7 ± 0.21 and 227 ± 0.48µg.mL⁻¹), respectively. In Gas chromatography-mass spectrometry (GC-MS) analysis, methanolic extract has 32 compounds, whereas; ethyl acetate and butanol fractions contain 40 and 36 compounds, respectively. Conclusion: Flaxseed contains totally different biologically active compounds that have been found to possess good variable activities, which can protect human body against several diseases.

Keywords: phenolic content, flavonoid content, HepG2, MCF7, hemolysis-assay, flaxseed

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Authors: Tomas Drevinskas, Ruta Mickiene, Audrius Maruska, Nicola Tiso, Algirdas Salomskas, Raimundas Lelesius, Agneta Karpovaite, Ona Ragazinskiene, Loreta Kubiliene

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Antiviral effect of substances accumulated by plants and natural products is known to ethno-pharmacy and modern day medicine. Antiviral properties are usually assigned to volatile compounds and polyphenols. This research work is divided into several parts and the task of this part was to investigate potential plants, potential substances and potential preparation conditions that can be used for the preparation of antiviral agents. Sixteen different medicinal plants, their parts and two types of propolis were selected for screening. Firstly, extraction conditions of non-volatile compounds were investigated: 3 pre-selected plants were extracted with 5 different ethanol – water mixtures (96%, 75%, 60%, 40%, 20 %, vol.) and bidistilled water. Total phenolic content, total flavonoid content and radical scavenging activity was determined. The results indicated that optimal extrahent is 40%, vol. of ethanol – water mixture. Further investigations were performed with the extrahent of 40%, vol. ethanol – water mixture. All 16 of selected plants, their parts and two types of propolis were extracted using selected extrahent. Determined total phenolic content, total flavonoid content and radical scavenging activity indicated that extracts of Origanum Vulgare L., Mentha piperita L., Geranium macrorrhizum L., Melissa officinalis L. and Desmodium canadence L. contains highest amount of extractable phenolic compounds (7.31, 5.48, 7.88, 8.02 and 7.16 rutin equivalents (mg/ ml) respectively), flavonoid content (2.14, 2.23, 2.49, 0.79 and 1.51 rutin equivalents (mg/ml) respectively) and radical scavenging activity (11.98, 8.72, 13.47, 13.22 and 12.22 rutin equivalents (mg/ml) respectively). Composition of the extracts is analyzed using HPLC.

Keywords: antiviral effect, plants, propolis, phenols

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Authors: Vanessa Roulet, Marc Turini, Juliane Hey, Stéphanie Bord-Romeu, Emilie Bonzom, Mahmoud Badawi, Mohammed-Amine Chakir, Valérie Simon, Vanessa Viotti, Jérémie Gautier, Françoise Le Boulaire, Catherine Coignard, Claire Vincent, Sandrine Greaume, Isabelle Voisin

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Background: Beckman Coulter, Inc. has recently developed fully automated assays for the detection of HBsAg on a new immunoassay platform. The objective of this European multicenter study was to evaluate the performance of the ACCESS HBsAg and ACCESS HBsAg Confirmatory assays† on the recently CE-marked DxI 9000 ACCESS Immunoassay Analyzer. Methods: The clinical specificity of the ACCESS HBsAg and HBsAg Confirmatory assays was determined using HBsAg-negative samples from blood donors and hospitalized patients. The clinical sensitivity was determined using presumed HBsAg-positive samples. Sample HBsAg status was determined using a CE-marked HBsAg assay (Abbott ARCHITECT HBsAg Qualitative II, Roche Elecsys HBsAg II, or Abbott PRISM HBsAg assay) and a CE-marked HBsAg confirmatory assay (Abbott ARCHITECT HBsAg Qualitative II Confirmatory or Abbott PRISM HBsAg Confirmatory assay) according to manufacturer package inserts and pre-determined testing algorithms. False initial reactive rate was determined on fresh hospitalized patient samples. The sensitivity for the early detection of HBV infection was assessed internally on thirty (30) seroconversion panels. Results: Clinical specificity was 99.95% (95% CI, 99.86 – 99.99%) on 6047 blood donors and 99.71% (95%CI, 99.15 – 99.94%) on 1023 hospitalized patient samples. A total of six (6) samples were found false positive with the ACCESS HBsAg assay. None were confirmed for the presence of HBsAg with the ACCESS HBsAg Confirmatory assay. Clinical sensitivity on 455 HBsAg-positive samples was 100.00% (95% CI, 99.19 – 100.00%) for the ACCESS HBsAg assay alone and for the ACCESS HBsAg Confirmatory assay. The false initial reactive rate on 821 fresh hospitalized patient samples was 0.24% (95% CI, 0.03 – 0.87%). Results obtained on 30 seroconversion panels demonstrated that the ACCESS HBsAg assay had equivalent sensitivity performances compared to the Abbott ARCHITECT HBsAg Qualitative II assay with an average bleed difference since first reactive bleed of 0.13. All bleeds found reactive in ACCESS HBsAg assay were confirmed in ACCESS HBsAg Confirmatory assay. Conclusion: The newly developed ACCESS HBsAg and ACCESS HBsAg Confirmatory assays from Beckman Coulter have demonstrated high clinical sensitivity and specificity, equivalent to currently marketed HBsAg assays, as well as a low false initial reactive rate. †Pending achievement of CE compliance; not yet available for in vitro diagnostic use. 2023-11317 Beckman Coulter and the Beckman Coulter product and service marks mentioned herein are trademarks or registered trademarks of Beckman Coulter, Inc. in the United States and other countries. All other trademarks are the property of their respective owners.

Keywords: dxi 9000 access immunoassay analyzer, hbsag, hbv, hepatitis b surface antigen, hepatitis b virus, immunoassay

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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: Hye Jin Kim, Hwa Sung Shin

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Ischemic stroke means the caused brain damage due to neurological defects, occurring occlusion of cerebral vascular resulting in thrombus or embolism. t-PA (tissue Plasminogen Activator) is the only thrombolytic agent passed the FDA (Food and Drug Administration). However, t-PA directly dissolves the thrombus (direct activity) through fibrinolysis, showing side effects such as re-occlusion. In this study, we evaluated the thrombolytic activities of the serine protease extracted from lugworms inhabiting tidal flats. The new serine protease identified as 38 kDa by SDS-PAGE was not toxic to brain endothelial cells line (hCMEC/D3). Also, the plasmin synthesis inhibition activity (indirect activity) of the new serine protease was confirmed through fibrin zymography assay and fibrin plate assay. It was higher than direct activity as compared to u-PA (urokinase Plasminogen Activator). The activities were found to be maintained at a wide range of temperature (4-70 ℃) and pH 7-10 compared to previous thrombolytic agents from the azocasein assay. In addition, the new serine protease has shown anticoagulant activity from fibrinogenolytic activity assay. In conclusion, the serine protease in lug worms inhabiting the tidal flats could be considered a promising thrombolytic candidate for the treatment of ischemic stroke.

Keywords: alkaline serine protease, bifunctional thrombolytic activity, fibrinolytic activity, ischemic stroke, lug worms

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Authors: F. Belkhiri, A. Baghiani, S. Boumerfeg, N. Charef, S. Khennouf, L. Arrar

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The present study was conducted to evaluate the in vitro antioxidant and antibacterial properties of methanolic extracts from roots of Tamus communis L. (TCRE), which is a plant used in traditional medicine in Algeria. The antioxidant potential of pattern was evaluated using tow complementary techniques, inhibition of free radical DPPH and the test of β-Carotene/linoleic acid. The antioxidant test indicates that non-polar fractions of TCRE (chloroform and ethyl acetate fractions) were more active than the polar fractions. Among these fractions, the chloroform extract appear in the DPPH test an IC50 of (18.89 µg/ml) comparable to that of BHT (18.6 µg/ml). This fraction was able to inhibiting the oxidation of β-Carotene with a percentage of inhibition (89.84 %). In antibacterial test, non-polar fractions showed antibacterial activity very important compared with the polar fractions. These fractions have inhibited the growth of four from nine bacterial strains, causing zones of inhibition from 08 to 23 mm of diameter.

Keywords: antioxidant activity, antibacterial activity, Tamus communis L., polar fractions

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Authors: Kyung Jin Kim, Jiwon Kwak, Jae-Hoon Lee, Soo Suk Lee

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MicroRNAs (miRNA) are a class of endogenous, single-stranded, small, and non-protein coding RNA molecules typically 20-25 nucleotides long. They are thought to regulate the expression of other genes in a broad range by binding to 3’- untranslated regions (3’-UTRs) of specific mRNAs. The detection of miRNAs is very important for understanding of the function of these molecules and in the diagnosis of variety of human diseases. However, detection of miRNAs is very challenging because of their short length and high sequence similarities within miRNA families. So, a simple-to-use, low-cost, and highly sensitive method for the detection of miRNAs is desirable. In this study, we demonstrate a novel bi-directional extension (BDE) assay. In the first step, a specific linear RT primer is hybridized to 6-10 base pairs from the 3’-end of a target miRNA molecule and then reverse transcribed to generate a cDNA strand. After reverse transcription, the cDNA was hybridized to the 3’-end which is BDE sequence; it played role as the PCR template. The PCR template was amplified in an SYBR green-based quantitative real-time PCR. To prove the concept, we used human brain total RNA. It could be detected quantitatively in the range of seven orders of magnitude with excellent linearity and reproducibility. To evaluate the performance of BDE assay, we contrasted sensitivity and specificity of the BDE assay against a commercially available poly (A) tailing method using miRNAs for let-7e extracted from A549 human epithelial lung cancer cells. The BDE assay displayed good performance compared with a poly (A) tailing method in terms of specificity and sensitivity; the CT values differed by 2.5 and the melting curve showed a sharper than poly (A) tailing methods. We have demonstrated an innovative, cost-effective BDE assay that allows improved sensitivity and specificity in detection of miRNAs. Dynamic range of the SYBR green-based RT-qPCR for miR-145 could be represented quantitatively over a range of 7 orders of magnitude from 0.1 pg to 1.0 μg of human brain total RNA. Finally, the BDE assay for detection of miRNA species such as let-7e shows good performance compared with a poly (A) tailing method in terms of specificity and sensitivity. Thus BDE proves a simple, low cost, and highly sensitive assay for various miRNAs and should provide significant contributions in research on miRNA biology and application of disease diagnostics with miRNAs as targets.

Keywords: bi-directional extension (BDE), microRNA (miRNA), poly (A) tailing assay, reverse transcription, RT-qPCR

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Authors: Anna Gramza-Michałowska, Dominik Kmiecik, Justyna Bilon, Joanna Skręty, Joanna Kobus-Cisowska, Józef Korczak, Andrzej Sidor

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Background: Hypertension is one of the civilization diseases with a global scope. Many research showed that every day diet influences significantly our health, helping with the prophylaxis and diseases treatment. The key factor here is the presence of plant origin natural bio active components. Aim: The following research describes snack health-oriented products for hypertension sufferers enriched with selected plant ingredients. Various analytical methods have been applied to determine product’s basic composition and their antioxidant activity. Methods: Snack products was formulated from a composition of different flours, oil, yeast, plant particles and extracts. Basic composition of a product was evaluated as content of protein, lipids, fiber, ash and caloricity. Antioxidant capacity of snacks was evaluated with use radical scavenging methods (DPPH, ABTS) and ORAC value. Proposed snacks as new product was also characterized with sensory analysis. Results and discussion: Results showed that addition of phyto nutrients allowed to improve nutritional and antioxidative value of examined products. Also the anti radical potential was significantly increased, with no loss of sensory value of a snacks. Conclusions: Designed snack is rich in polyphenolics, that express high antioxidant activity, helpful in hypertension and as low calories product obesity prophylaxis.

Keywords: antioxidant, well-being, hypertension, bioactive compounds

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Authors: Kai-Cheng Yang, Yen-Ling Chen, Er-Chieh Cho, Kuen-Chan Lee

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Titanium dioxide nanomaterials offer superior protection for human skin against the full spectrum of ultraviolet light. However, some literature reviews indicated that it might be associated with adverse effects such as cytotoxicity or reactive oxygen species (ROS) due to their nanoscale. The surface of fullerene is covered with π electrons constituting aromatic structures, which can effectively scavenge large amount of radicals. Unfortunately, fullerenes are poor solubility in water, severe aggregation, and toxicity in biological applications when dispersed in solvent have imposed the limitations to the use of fullerenes. Carboxyfullerene acts as the scavenger of radicals for several years. Some reports indicate that carboxyfullerene not only decrease the concentration of free radicals in ambience but also prevent cells from reducing the number or apoptosis under UV irradiation. The aim of this study is to decorate fullerene –C70-carboxylic acid (C70-COOH) on the surface of titanium dioxide nanoparticles (P25) for the purpose of scavenging ROS during the irradiation. The modified material is prepared through the esterification of C70-COOH with P25 (P25/C70-COOH). The binding edge and structure are studied by using Transmission electron microscope (TEM) and Fourier transform infrared (FTIR). The diameter of P25 is about 30 nm and C70-COOH is found to be conjugated on the edge of P25 in aggregation morphology with the size of ca. 100 nm. In the next step, the FTIR was used to confirm the binding structure between P25 and C70-COOH. There are two new peaks are shown at 1427 and 1720 cm-1 for P25/C70-COOH, resulting from the C–C stretch and C=O stretch formed during esterification with dilute sulfuric acid. The IR results further confirm the chemically bonded interaction between C70-COOH and P25. In order to provide the evidence of scavenging radical ability of P25/C70-COOH, we chose pyridoxine (Vit.B6) and terephthalic acid (TA) to react with singlet oxygen and hydroxyl radicals. We utilized these chemicals to observe the radicals scavenging statement via detecting the intensity of ultraviolet adsorption or fluorescence emission. The UV spectra are measured by using different concentration of C70-COOH modified P25 with 1mM pyridoxine under UV irradiation for various duration times. The results revealed that the concentration of pyridoxine was increased when cooperating with P25/C70-COOH after three hours as compared with control (only P25). It indicates fewer radicals could be reacted with pyridoxine because of the absorption via P25/C70-COOH. The fluorescence spectra are observed by measuring P25/C70-COOH with 1mM terephthalic acid under UV irradiation for various duration times. The fluorescence intensity of TAOH was decreased in ten minutes when cooperating with P25/C70-COOH. Here, it was found that the fluorescence intensity was increased after thirty minutes, which could be attributed to the saturation of C70-COOH in the absorption of radicals. However, the results showed that the modified P25/C70-COOH could reduce the radicals in the environment. Therefore, we expect that P25/C70-COOH is a potential materials in using for antioxidant.

Keywords: titanium dioxide, fullerene, radical scavenging activity, antioxidant

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Authors: Muhammad Sajid Arshad, Sadaf Bashir

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In this study, the effects of pomegranate peel based edible coating were determined on safety and quality of chicken nuggets. Four treatment groups were prepared as control (without coating), coating with sodium alginate (SA) (1.5%), pomegranate peel powder (PPP) (1.5%), and combination of SA and PPP. There was a significant variation observed with respect to coating treatments and storage intervals. The chicken nuggets were subjected to refrigerated storage (40C) and were analyzed at regular intervals of 0, 7, 14 1 and 21 days. The microbiological quality was determined by total aerobic and coliform counts. Total aerobic (5.09±0.05 log CFU/g) and coliforms (3.91±0.06 log CFU/g) counts were higher in uncoated chicken nuggets whereas lower was observed in coated chicken nuggets having combination of SA and PPP. Likewise, antioxidants potential of chicken nuggets was observed by assessing total phenolic contents (TPC) and DPPH activity. Higher TPC (135.66 GAE/100g) and DPPH (64.65%) were found in combination with SA and PPP, whereas minimum TPC (91.38) and DPPH (41.48) was observed in uncoated chicken nuggets. Regarding the stability analysis of chicken nuggets, thiobarbituric acid reactive substances (TBARS) and peroxide value (POV) were determined. Higher TBARS (1.62±0.03 MDA/Kg) and POV (0.92±0.03 meq peroxide/kg) were found in uncoated chicken nuggets. Hunter color values were also observed in both uncoated and coated chicken nuggets. Sensorial attributes were also observed by the trained panelists. The higher sensory score for appearance, color, taste, texture and overall acceptability were observed in control (uncoated) while in coated treatments, it was found within acceptable limits. In nutshell, the combination of SA and PPP enhanced the overall quality, antioxidant potential, and stability of chicken nuggets.

Keywords: chicken nuggets, edible coatings, pomegranate peel powder, sodium alginate

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Authors: Christina Drosou, Konstantina E. Kyriakopoulou, Andreas Bimpilas, Dimitrios Tsimogiannis, Magdalini C. Krokida

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Rationale: Agiorgitiko is one of the most widely-grown and commercially well-established red wine varieties in Greece. Each year viticulture industry produces a large amount of waste consisting of grape skins and seeds (pomace) during a short period. Grapes contain polyphenolic compounds which are partially transferred to wine during winemaking. Therefore, winery wastes could be an alternative cheap source for obtaining such compounds with important antioxidant activity. Specifically, red grape waste contains anthocyanins and flavonols which are characterized by multiple biological activities, including cardioprotective, anti-inflammatory, anti-carcinogenic, antiviral and antibacterial properties attributed mainly to their antioxidant activity. Ultrasound assisted extraction (UAE) is considered an effective way to recover phenolic compounds, since it combines the advantage of mechanical effect with low temperature. Moreover, green solvents can be used in order to recover extracts intended for used in the food and nutraceutical industry. Apart from the extraction, pre-treatment process like drying can play an important role on the preservation of the grape pomace and the enhancement of its antioxidant capacity. Objective: The aim of this study is to recover natural extracts from winery waste with high antioxidant capacity using green solvents so they can be exploited and utilized as enhancers in food or nutraceuticals. Methods: Agiorgitiko grape pomace was dehydrated by air drying (AD) and accelerated solar drying (ASD) in order to explore the effect of the pre-treatment on the recovery of bioactive compounds. UAE was applied in untreated and dried samples using water and water: ethanol (1:1) as solvents. The total antioxidant potential and phenolic content of the extracts was determined using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay and Folin-Ciocalteu method, respectively. Finally, the profile of anthocyanins and flavonols was specified using HPLC-DAD analysis. The efficiency of processes was determined in terms of extraction yield, antioxidant activity, phenolic content and the anthocyanins and flavovols profile. Results & Discussion: The experiments indicated that the pre-treatment was essential for the recovery of highly nutritious compounds from the pomace as long as the extracts samples showed higher phenolic content and antioxidant capacity. Water: ethanol (1:1) was considered a more effective solvent on the recovery of phenolic compounds. Moreover, ASD grape pomace extracted with the solvent system exhibited the highest antioxidant activity (IC50=0.36±0.01mg/mL) and phenolic content (TPC=172.68±0.01mgGAE/g dry extract), followed by AD and untreated pomace. The major compounds recovered were malvidin3-O-glucoside and quercetin3-O-glucoside according to the HPLC analysis. Conclusions: Winery waste can be exploited for the recovery of nutritious compounds using green solvents such as water or ethanol. The pretreatment of the pomace can significantly affect the concentration of phenolic compounds, while UAE is considered a highly effective extraction process.

Keywords: agiorgitico grape pomace, antioxidants, phenolic compounds, ultrasound assisted extraction

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Authors: Chiagoziem A. Otuechere, Ebenezer O. Farombi

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The protective effect of dichloromethane: methanol extract of Pterocarpus mildbraedii (PME), a widely consumed Nigerian leafy vegetable, on the toxicity of propanil was investigated in male rats. Animals were distributed into eight groups of five each. Group 1 served as control and received normal saline while rats in groups 2, 3, and 4 received 100 mg/kg, 200 mg/kg, and 400 mg/kg extract doses respectively. Group 5 rats were orally administered 200 mg/kg propanil while groups 6, 7, and 8 rats were given propanil plus extract. Oral administration of propanil elicited a 14.8%, 5%, 122%, and 78% increase in the activity of serum enzymes; alanine aminotransferase (AST), alanine aminotransferase(ALT), Alkaline phoshatase (ALP) and Gamma glutamyl transferase (ﻻGT). There were also increase in Lactate dehydrogenase (LDH) activity, direct bilirubin and lipid peroxidation levels. Furthermore, PME significantly attenuated the marked hepatic oxidative damage that accompanied propanil treatment. The extract significantly decreased LDH activity and bilirubin levels following propanil treatment. Furthermore, propanil-induced alterations in the activities of antioxidant enzymes: Superoxide dismutase (SOD), catalase (CAT) and glutathione s-transferase (GST) in these rats were modulated by the extract. The percentage DPPH Radical Scavenging Activity of the extract was determined as 55% and compared to those of Gallic acid (49%). Hepatic histology examination further confirmed the damage to the liver as it revealed severe periportal cellular infiltration of the hepatocytes. These biochemical and morphological alterations were attenuated in rats pre-treated with 100 mg/kg and 200 mg/kg doses of the extract. These results suggest that PME possesses protective effect against propanil-induced hepatotoxicity.

Keywords: antioxidant, hepatoprotection, Pterocarpus mildbraedii, propanil

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Authors: Esma Hande Alıcı, Gülnur Arabacı

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Antioxidants are compounds or systems that can safely interact with free radicals and terminate the chain reaction before vital molecules are damaged. The anti-oxidative effectiveness of these compounds depends on their chemical characteristics and physical location within a food (proximity to membrane phospholipids, emulsion interfaces, or in the aqueous phase). Antioxidants (e.g., flavonoids, phenolic acids, tannins, vitamin C, vitamin E) have diverse biological properties, such as antiinflammatory, anti-carcinogenic and anti-atherosclerotic effects, reduce the incidence of coronary diseases and contribute to the maintenance of gut health by the modulation of the gut microbial balance. Plants are excellent sources of antioxidants especially with their high content of phenolic compounds. Raphanus raphanistrum L., the wild radish, is a flowering plant in the family Brassicaceae. It grows in Asia and Mediterranean region. It has been introduced into most parts of the world. It spreads rapidly, and is often found growing on roadsides or in other places where the ground has been disturbed. It is an edible plant, in Turkey its fresh aerial parts are mostly consumed as a salad with olive oil and lemon juice after boiled. The leaves of the plant are also used as anti-rheumatic in traditional medicine. In this study, we determined the antioxidant capacity of two different solvent fractions (methanol and ethyl acetate) obtained from Raphanus raphanistrum L. plant leaves. Antioxidant capacity of the plant was introduced by using three different methods: DPPH radical scavenging activity, CUPRAC (Cupric Ion Reducing Antioxidant Capacity) activity and Reducing power activity.

Keywords: antioxidant activity, antioxidant capacity, Raphanis raphanistrum L., wild radish

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Authors: Afifah Nur Aini, Elsa Mega Suryani, Betty Lukiaty

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Drymoglossum piloselloides or more commonly known as sisik naga fern is a member of Polipodiaceae Family that is abundant and widely distributed in nature. That being said, there hasn’t been many studies reporting about the benefits of this fern. The aim of this study was to find out the active compounds and antioxidant activity of water fraction extract of sisik naga leaves. The study will be able to optimize the use of this fern in the future. In this study, phytochemical test was done qualitatively by using Mayer, Dragendorff and Wagner reagent for alkaloid test; FeCl3 for phenolic test; Shinoda test for flavonoid; Liebermann-Burchard test for triterprnoid and Forth test for saponin. Antioxidant activity test was done by using 20D spectronic spectrophotometer to determine the percentage of DPPH free radical inhibition. The results showed that water fraction extract of sisik naga leaves contain phenolic and IC50 = 5.44 μg/ml. This means that sisik naga leaves can be used as an antioxidant.

Keywords: antioxidant activity test, dpph, phytochemical test, drymoglossum piloselloides

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Authors: Dan W. Rhodes, Juliane Hey, Magali Karagueuzian, Florianne Martinez, Yael Sandowski, Vanessa Roulet, Mahmoud Badawi, Mohammed-Amine Chakir, Valérie Simon, Jérémie Gautier, Françoise Le Boulaire, Catherine Coignard, Claire Vincent, Sandrine Greaume, Isabelle Voisin

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Background: Beckman Coulter, Inc. (BEC) has recently developed a fully automated second-generation anti-HCV test on a new immunoassay platform. The objective of this multicenter study conducted in Europe was to evaluate the performance of the ACCESS anti-HCV assay on the recently CE-marked DxI 9000 ACCESS Immunoassay Analyzer as an aid in the diagnosis of HCV (Hepatitis C Virus) infection and as a screening test for blood and plasma donors. Methods: The clinical specificity of the ACCESS anti-HCV assay was determined using HCV antibody-negative samples from blood donors and hospitalized patients. Sample antibody status was determined by a CE-marked anti-HCV assay (Abbott ARCHITECTTM anti-HCV assay or Abbott PRISM HCV assay) with an additional confirmation method (Immunoblot testing with INNO-LIATM HCV Score - Fujirebio), if necessary, according to pre-determined testing algorithms. The clinical sensitivity was determined using known HCV antibody-positive samples, identified positive by Immunoblot testing with INNO-LIATM HCV Score - Fujirebio. HCV RNA PCR or genotyping was available on all Immunoblot positive samples for further characterization. The false initial reactive rate was determined on fresh samples from blood donors and hospitalized patients. Thirty (30) commercially available seroconversion panels were tested to assess the sensitivity for early detection of HCV infection. The study was conducted from November 2019 to March 2022. Three (3) external sites and one (1) internal site participated. Results: Clinical specificity (95% CI) was 99.7% (99.6 – 99.8%) on 5852 blood donors and 99.0% (98.4 – 99.4%) on 1527 hospitalized patient samples. There were 15 discrepant samples (positive on ACCESS anti-HCV assay and negative on both ARCHITECT and Immunoblot) observed with hospitalized patient samples, and of note, additional HCV RNA PCR results showed five (5) samples had positive HCV RNA PCR results despite the absence of HCV antibody detection by ARCHITECT and Immunoblot, suggesting a better sensitivity of the ACCESS anti-HCV assay with these five samples compared to the ARCHITECT and Immunoblot anti-HCV assays. Clinical sensitivity (95% CI) on 510 well-characterized, known HCV antibody-positive samples was 100.0% (99.3 – 100.0%), including 353 samples with known HCV genotypes (1 to 6). The overall false initial reactive rate (95% CI) on 6630 patient samples was 0.02% (0.00 – 0.09%). Results obtained on 30 seroconversion panels demonstrated that the ACCESS anti-HCV assay had equivalent sensitivity performances, with an average bleed difference since the first reactive bleed below one (1), compared to the ARCHITECTTM anti-HCV assay. Conclusion: The newly developed ACCESS anti-HCV assay from BEC for use on the DxI 9000 ACCESS Immunoassay Analyzer demonstrated high clinical sensitivity and specificity, equivalent to currently marketed anti-HCV assays, as well as a low false initial reactive rate.

Keywords: DxI 9000 ACCESS Immunoassay Analyzer, HCV, HCV antibody, Hepatitis C virus, immunoassay

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Authors: Shrestha Sharma, Jasjeet K. Sahni, Javed Ali, Sanjula Baboota

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Introduction: Rutin is a naturally occurring strong antioxidant molecule belonging to bioflavonoid category. Due to its free radical scavenging properties, it has been found to be beneficial in the treatment of various diseases including inflammation, cancer, diabetes, allergy, cardiovascular disorders and various types of microbial infections. Despite its beneficial effects, it suffers from the problem of low aqueous solubility which is responsible for low oral bioavailability. The aim of our study was to optimize and characterize self-nanoemulsifying drug delivery system (SNEDDS) of rutin using Box-Behnken design (BBD) combined with a desirability function. Further various antioxidant, pharmacokinetic and pharmacodynamic studies were performed for the optimized rutin SNEDDS formulation. Methodologies: Selection of oil, surfactant and co-surfactant was done on the basis of solubility/miscibility studies. Sefsol+ Vitamin E, Solutol HS 15 and Transcutol P were selected as oil phase, surfactant and co-surfactant respectively. Optimization of SNEDDS formulations was done by a three-factor, three-level (33)BBD. The independent factors were Sefsol+ Vitamin E, Solutol HS15, and Transcutol P. The dependent variables were globule size, self emulsification time (SEF), % transmittance and cumulative percentage drug released. Various response surface graphs and contour plots were constructed to understand the effect of different factor, their levels and combinations on the responses. The optimized Rutin SNEDDS formulation was characterized for various parameters such as globule size, zeta potential, viscosity, refractive index , % Transmittance and in vitro drug release. Ex vivo permeation studies and pharmacokinetic studies were performed for optimized formulation. Antioxidant activity was determined by DPPH and reducing power assays. Anti-inflammatory activity was determined by using carrageenan induced rat paw oedema method. Permeation of rutin across small intestine was assessed using confocal laser scanning microscopy (CLSM). Major findings:The optimized SNEDDS formulation consisting of Sefsol+ Vitamin E - Solutol HS15 -Transcutol HP at proportions of 25:35:17.5 (w/w) was prepared and a comparison of the predicted values and experimental values were found to be in close agreement. The globule size and PDI of optimized SNEDDS formulation was found to be 16.08 ± 0.02 nm and 0.124±0.01 respectively. Significant (p˂0.05) increase in percentage drug release was achieved in the case of optimized SNEDDS formulation (98.8 %) as compared to rutin suspension. Furthermore, pharmacokinetic study showed a 2.3-fold increase in relative oral bioavailability compared with that of the suspension. Antioxidant assay results indicated better efficacy of the developed formulation than the pure drug and it was found to be comparable with ascorbic acid. The results of anti-inflammatory studies showed 72.93 % inhibition for the SNEDDS formulation which was significantly higher than the drug suspension 46.56%. The results of CLSM indicated that the absorption of SNEDDS formulation was considerably higher than that from rutin suspension. Conclusion: Rutin SNEDDS have been successfully prepared and they can serve as an effective tool in enhancing oral bioavailability and efficacy of Rutin.

Keywords: rutin, oral bioavilability, pharamacokinetics, pharmacodynamics

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Authors: Weena Siangphro, Orawon Chailapakul, Kriangsak Songsrirote

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A simple, rapid, sensitive, and economical method based on colorimetry for the determination of paraquat, a widely used herbicide, was developed. Citrate-coated silver nanoparticles (AgNPs) were synthesized as colorimetric probe. The mechanism of the assay is related to aggregation of negatively charged AgNPs induced by positively-charged paraquat resulting from coulombic attraction which causes the color change from deep greenish yellow to pale yellow upon the concentrations of paraquat. Silica gel was exploited as paraquat adsorbent for purification and pre-concentration prior to the direct determination with negatively charged AgNPs without elution step required. The validity of the proposed approach was evaluated by spiking standard paraquat in water and plant samples. Recoveries of paraquat in water samples were 93.6-95.4%, while those in plant samples were 86.6-89.5% by using the optimized extraction procedure. The absorbance of AgNPs at 400 nm was linearly related to the concentration of paraquat over the range of 0.05-50 mg/L with detection limits of 0.05 ppm for water samples, and 0.10 ppm for plant samples.

Keywords: colorimetric assay, paraquat, silica gel, silver nanoparticles

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