Search results for: no-volatils extracts
Commenced in January 2007
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Edition: International
Paper Count: 947

Search results for: no-volatils extracts

617 The Hidden Mechanism beyond Ginger (Zingiber officinale Rosc.) Potent in vivo and in vitro Anti-Inflammatory Activity

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

Abstract:

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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616 Formulation and Characterization of Antimicrobial Herbal Mouthwash from Some Herbal Extracts for Treatment of Periodontal Diseases

Authors: Reenu Yadav, Abhay Asthana, S. K. Yadav

Abstract:

Purpose: The aim of the present work was to develop an oral gel for brushing with an antimicrobial activity which will cure/protect from various periodontal diseases such as periodontitis, gingivitis, and pyorrhea. Methods: Plant materials procured from local suppliers, extracted and standardized. Screening of antimicrobial activity was carried out with the help of disk diffusion method. The gel was formulated by dried extracts of Beautea monosperma and Cordia obliquus. Gels were evaluated on various parameters and standardization of the formulation was performed. The release of drugs was studied in pH 6.8 using a mastication device.Total phenolic and flavonoid contents were estimated by folin-Ciocalteu and aluminium chloride method, and stability studies were performed (40°C and RH 75% ± 5% for 90 days) to assess the effect of temperature and humidity on the concentration of phenolic and flavonoid contents. The results of accelerated stability conditions were compared with that of samples kept at controlled conditions (RT). The control samples were kept at room temperature (25°C, 35% RH for 180 days). Results: Results are encouraging; extracts possess significant antimicrobial activity at very low concentration (15µg/disc, 20µg/disc and 15 µg/ disc) on oral pathogenic bacteria. The formulation has optimal characteristics, as well as has a pleasant appearance, fragrance, texture, and taste, is highly acceptable by the volunteers. The diffusion coefficient values ranged from 0.6655 to 0.9164. Since the R values of korsmayer papas were close to 1, Drug release from formulation follows matrix diffusion kinetics. Hence, diffusion was the mechanism of the drug release. Formulation follows non-Fickian transport mechanism. Most Formulations released 50 % of their contents within 25-30 minutes. Results obtained from the accelerated stability studies are indicative of a slight reduction in flavonoids and phenolic contents with time on long time storage. When measured degradation under ambient conditions, degradation was significantly lower than in accelerated stability study. Conclusion: Plant extracts possess compounds with antimicrobial properties can be used as. Developed formulation will cure/protect from various periodontal diseases. Further development and evaluations oral gel including the isolated compounds on the commercial scale and their clinical and toxicological studies are the future challenges.

Keywords: herbal gel, dental care, ambient conditions, commercial scale

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615 Evaluation of Anti-inflammatory Activities of Extracts Obtained from Capparis Erythrocarpos In-Vivo

Authors: Benedict Ofori, Kwabena Sarpong, Stephen Antwi

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Background: Medicinal plants are utilized all around the world and are becoming increasingly important economically. The WHO notes that ‘inappropriate use of traditional medicines or practices can have negative or dangerous effects and that future research is needed to ascertain the efficacy and safety of such practices and medicinal plants used by traditional medicine systems. The poor around the world have limited access to palliative care or pain relief. Pharmacologists have been focused on developing safe and effective anti-inflammatory drugs. Most of the issues related to their use have been linked to the fact that numerous traditional and herbal treatments are classified in different nations as meals or dietary supplements. As a result, there is no need for evidence of the quality, efficacy, or safety of these herbal formulations before they are marketed. The fact that access to drugs meant for pain relief is limited in low-income countries means advanced studies should be done on home drugs meant for inflammation to close the gap. Methods: The ethanolic extracts of the plant were screened for the presence of 10 phytochemicals. The Pierce BCA Protein Assay Kit was used for the determination of the protein concentration of the egg white. The rats were randomly selected and put in 6 groups. The egg white was sub-plantar injected into the right-hand paws of the rats to induce inflammation. The animals were treated with the three plant extracts obtained from the root bark, stem, and leaves of the plant. The control groups were treated with normal saline, while the standard groups were treated with standard drugs indomethacin and celecoxib. Plethysmometer was used to measure the change in paw volume of the animals over the course of the experiment. Results: The results of the phytochemical screening revealed the presence of reducing sugars and saponins. Alkaloids were present in only R.L.S (1:1:1), and phytosterols were found in R.L(1:1) and R.L.S (1:1:1). The estimated protein concentration was found to be 103.75 mg/ml. The control group had an observable increase in paw volume, which indicated that inflammation was induced during the 5 hours. The increase in paw volume for the control group peaked at the 1st hour and decreased gradually throughout the experiment, with minimal changes in the paw volumes. The 2nd and 3rd groups were treated with 20 mg/kg of indomethacin and celecoxib. The anti-inflammatory activities of indomethacin and celecoxib were calculated to be 21.4% and 4.28%, respectively. The remaining 3 groups were treated with 2 dose levels of 200mg/kg plant extracts. R.L.S, R.L, and S.R.L had anti-inflammatory activities of 22.3%, 8.2%, and 12.07%, respectively. Conclusions: Egg albumin-induced paw model in rats can be used to evaluate the anti-inflammatory activity of herbs that might have potential anti-inflammatory activity. Herbal medications have potential anti-inflammatory activities and can be used to manage various inflammatory conditions if their efficacy and side effects are well studied. The three extracts all possessed anti-inflammatory activity, with R.L.S having the highest anti-inflammatory activity.

Keywords: inflammation, capparis erythrocarpos, anti-inflammatory activity, herbal medicine, paw volume, egg albumin

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614 The Application of Whole-Cell Luminescent Biosensors for Assessing Bactericidal Properties of Medicinal Plants

Authors: Yuliya Y. Gavrichenko

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Background and Aims: The increasing bacterial resistance to almost all the available antibiotics has encouraged scientists to search for alternative sources of antibacterial agents. Nowadays, it is known that many plant secondary metabolites have diverse biological activity. These compounds can be potentially active against human bacterial and viral infections. Extended research has been carried out to explore the use of the luminescent bacterial test as a rapid, accurate and inexpensive method to assess the antibacterial properties and to predict the biological activity spectra for plant origin substances. Method: Botanical material of fifteen species was collected from their natural and cultural habitats on the Crimean peninsula. The aqueous extracts of following plants were tested: Robinia pseudoacacia L., Sideritis comosa, Cotinus coggygria Scop., Thymus serpyllum L., Juglans regia L., Securigera varia L., Achillea millefolium L., Phlomis taurica, Corylus avellana L., Sambucus nigra L., Helichrysum arenarium L., Glycyrrhiza glabra L., Elytrigia repens L., Echium vulgare L., Conium maculatum L. The test was carried out using luminous strains of marine bacteria Photobacterium leiognathi, which was isolated from the Sea of Azov as well as four Escherichia coli MG1655 recombinant strains harbouring Vibrio fischeri luxCDABE genes. Results: The bactericidal capacity of plant extracts showed significant differences in the study. Cotinus coggygria, Phlomis taurica, Juglans regia L. proved to be the most toxic to P. leiognathi. (EC50 = 0.33 g dried plant/l). Glycyrrhiza glabra L., Robinia pseudoacacia L., Sideritis comosa and Helichrysum arenarium L. had moderate inhibitory effects (EC50 = 3.3 g dried plant/l). The rest of the aqueous extracts have decreased the luminescence of no more than 50% at the lowest concentration (16.5 g dried plant/l). Antibacterial activity of herbal extracts against constitutively luminescent E. coli MG1655 (pXen7-lux) strain was observed at approximately the same level as for P. leiognathi. Cotinus coggygria and Conium maculatum L. extracts have increased light emission in the mutant E. coli MG1655 (pFabA-lux) strain which is associated with cell membranes damage. Sideritis comosa, Phlomis taurica, Juglans regia induced SOS response in E. coli (pColD-lux) strain. Glycyrrhiza glabra L. induced protein damage response in E. coli MG1655 (pIbpA-lux) strain. Conclusion: The received results have shown that the plants’ extracts had nonspecific antimicrobial effects against both E. coli (pXen7-lux) and P. leiognathi biosensors. Mutagenic, cytotoxic and protein damage effects have been observed. In general, the bioluminescent inhibition test result correlated with the traditional use of screened plants. It leads to the following conclusion that whole-cell luminescent biosensors could be the indicator of overall plants antibacterial capacity. The results of the investigation have shown a possibility of bioluminescent method in medicine and pharmacy as an approach to research the antibacterial properties of medicinal plants.

Keywords: antibacterial property, bioluminescence, medicinal plants, whole-cell biosensors

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613 The Use of Antioxidant and Antimicrobial Properties of Plant Extracts for Increased Safety and Sustainability of Dairy Products

Authors: Loreta Serniene, Dalia Sekmokiene, Justina Tomkeviciute, Lina Lauciene, Vaida Andruleviciute, Ingrida Sinkeviciene, Kristina Kondrotiene, Neringa Kasetiene, Mindaugas Malakauskas

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One of the most important areas of product development and research in the dairy industry is the product enrichment with active ingredients as well as leading to increased product safety and sustainability. The most expanding field of the active ingredients is the various plants' CO₂ extracts with aromatic, antioxidant and antimicrobial properties. In this study, 15 plant extracts were evaluated based on their antioxidant, antimicrobial properties as well as sensory acceptance indicators for the development of new dairy products. In order to increase the total antioxidant capacity of the milk products, it was important to determine the content of phenolic compounds and antioxidant activity of CO₂ extract. The total phenolic content of fifteen different commercial CO₂ extracts was determined by the Folin-Ciocalteu reagent and expressed as milligrams of the Gallic acid equivalents (GAE) in gram of extract. The antioxidant activities were determined by 2.2′-azinobis-(3-ethylbenzthiazoline)-6-sulfonate (ABTS) methods. The study revealed that the antioxidant activities of investigated CO₂ extract vary from 4.478-62.035 µmole Trolox/g, while the total phenolic content was in the range of 2.021-38.906 mg GAE/g of extract. For the example, the estimated antioxidant activity of Chinese cinnamon (Cinammonum aromaticum) CO₂ extract was 62.023 ± 0.15 µmole Trolox/g and the total flavonoid content reached 17.962 ± 0.35 mg GAE/g. These two parameters suggest that cinnamon could be a promising supplement for the development of new cheese. The inhibitory effects of these essential oils were tested by using agar disc diffusion method against pathogenic bacteria, most commonly found in dairy products. The obtained results showed that essential oil of lemon myrtle (Backhousia citriodora) and cinnamon (Cinnamomum cassia) has antimicrobial activity against E. coli, S. aureus, B. cereus, P. florescens, L. monocytogenes, Br. thermosphacta, P. aeruginosa and S. typhimurium with the diameter of inhibition zones variation from 10 to 52 mm. The sensory taste acceptability of plant extracts in combination with a dairy product was evaluated by a group of sensory evaluation experts (31 individuals) by the criteria of overall taste acceptability in the scale of 0 (not acceptable) to 10 (very acceptable). Each of the tested samples included 200g grams of natural unsweetened greek yogurt without additives and 1 drop of single plant extract (essential oil). The highest average of overall taste acceptability was defined for the samples with essential oils of orange (Citrus sinensis) - average score 6.67, lemon myrtle (Backhousia citriodora) – 6.62, elderberry flower (Sambucus nigra flos.) – 6.61, lemon (Citrus limon) – 5.75 and cinnamon (Cinnamomum cassia) – 5.41, respectively. The results of this study indicate plant extracts of Cinnamomum cassia and Backhousia citriodora as a promising additive not only to increase the total antioxidant capacity of the milk products and as alternative antibacterial agent to combat pathogenic bacteria commonly found in dairy products but also as a desirable flavour for the taste pallet of the consumers with expressed need for safe, sustainable and innovative dairy products. Acknowledgment: This research was funded by the European Regional Development Fund according to the supported activity 'Research Projects Implemented by World-class Researcher Groups' under Measure No. 01.2.2-LMT-K-718.

Keywords: antioxidant properties, antimicrobial properties, cinnamon, CO₂ plant extracts, dairy products, essential oils, lemon myrtle

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612 In Vivo Maltase and Sucrase Inhibitory Activities of Five Underutilized Nigerian Edible Fruits

Authors: Mohammed Auwal Ibrahim, Isa Yunusa, Nafisa Kabir, Shazali Ali Baba, Amina Muhammad Yushau, Suraj Suraj Ibrahim, Zaharaddeen Idris Bello, Suleiman Haruna Suleiman, Murtala Bindawa Isah

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Background: Inhibition of intestinal maltase and sucrase prevents postprandial blood glucose excursions which are beneficial in ameliorating diabetes-associated complications. Objective: In this study, the inhibitory effects of fruit extracts of Parinari macrophylla, Detarium microcarpum, Ziziphus spina-christi, Z. mairei and Parkia biglobosa were investigated against intestinal maltase and sucrase. Methods: Rats were given co-administration of the fruit extracts with maltose or sucrose and blood glucose levels were measured at 0, 30, 90 and 120 min. Results: The glucose-time curves indicated that all the fruits had the most potent inhibitory effects on both maltase and sucrase within the first 30 min. The computed Area Under the Curves (AUC0-120)for all the fruits indicated more potent inhibitory effects against intestinal maltase than sucrase.The ED50 range for the fruits extract against maltase and sucrase were 647.15-1118.35 and 942.44-1851.94 mg/kg bw respectively. Conclusion: The data suggests that the fruits could prevent postprandial hyperglycemia via inhibition of intestinal maltase and sucrase.

Keywords: diabetes mellitus, fruits, α-glucosidases, maltase, sucrase

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611 A Scientific Method of Drug Development Based on Ayurvedic Bhaishajya Knowledge

Authors: Rajesh S. Mony, Vaidyaratnam Oushadhasala

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An attempt is made in this study to evolve a drug development modality based on classical Ayurvedic knowledge base as well as on modern scientific methodology. The present study involves (a) identification of a specific ailment condition, (b) the selection of a polyherbal formulation, (c) deciding suitable extraction procedure, (d) confirming the efficacy of the combination by in-vitro trials and (e) fixing up the recommended dose. The ailment segment selected is arthritic condition. The selected herbal combination is Kunturushka, Vibhitaki, Guggulu, Haridra, Maricha and Nirgundi. They were selected as per Classical Ayurvedic references, Authentified as per API (Ayurvedic Pharmacopeia of India), Extraction of each drug was done by different ratios of Hydroalcoholic menstrums, Invitro assessment of each extract after removing residual solvent for anti-Inflammatory, anti-arthritic activities (by UV-Vis. Spectrophotometer with positive control), Invitro assessment of each extract for COX enzyme inhibition (by UV-Vis. Spectrophotometer with positive control), Selection of the extracts was made having good in-vitro activity, Performed the QC testing of each selected extract including HPTLC, that is the in process QC specifications, h. Decision of the single dose with mixtures of selected extracts was made as per the level of in-vitro activity and available toxicology data, Quantification of major groups like Phenolics, Flavonoids, Alkaloids and Bitters was done with both standard Spectrophotometric and Gravimetric methods, Method for Marker assay was developed and validated by HPTLC and a good resolved HPTLC finger print was developed for the single dosage API (Active Pharmaceutical Ingredient mixture of extracts), Three batches was prepared to fix the in process and API (Active Pharmaceutical Ingredient) QC specifications.

Keywords: drug development, antiinflammatory, quality stardardisation, planar chromatography

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610 Walnut (Juglans Regia) Extracts: Investigation of Antioxidant Effect, Total Phenols and Tyrosinase Inhibitory Activity

Authors: N. Saki, S. Nalbantoglu, M. Akin, G. Arabaci

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Walnut has a great range of phenolic profile and it is used in Asia and Africa for treatment of many diseases and cancer. Phenolic compounds play a number of crucial roles in complex metabolism of plants and of also fruit trees. Consumption of certain phenolics in the food is considered beneficial for human nutrition. Phenolic compounds known as anti-radical inactivators with their high antioxidant activities and these activities play an important role in inhibition of multi-metal corrosion. Many common corrosion inhibitors that are still in use today are health hazards. Therefore, there is still an increased attention directed towards the development of environmentally compatible, nonpolluting corrosion inhibitors. The present study reports the total phenols content, antioxidant potentials and tyrosinase inhibitory activity of the walnut (Juglans regia L.) produced in Turkey. The anti-tyrosinase activity was investigated for walnut at 2 h extraction time and all extracts exhibited tyrosinase activity. The results of this study suggested that walnut can be used as an excellent, easily accessible source of natural antioxidant.

Keywords: antioxidant activity, Juglans Regia, total phenols, tyrosinase activity

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609 Chemical Characterization and Antioxidant Capacity of Flour From Two Soya Bean Cultivars (Glycine Max)

Authors: Meziani Samira, Menadi Noreddine, Labga Lahouaria, Chenni Fatima Zohra, Toumi Asma

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A comparative study between two varieties of soya beans was carried out in this work. The method consists of studying and proceeding to prepare a by-product (Flour) from two varieties of soybeans, a Chinese variety imported and marketed in Algeria. The chemical composition of ash, protein and fat was determined in this study. The minerals, namely potassium and sodium, were measured by flame spectrophotometer. In addition, the estimation of the polyphenol content and evaluation of the antioxidant activity Ferric Reducing Antioxidant Power assay (FRAP) f the methanol extracts of the flours were also carried out. The result revealed that soy flour from two cultivars, on average, contained 8% moisture, more than 50% protein, 1.58-1.87g fat, and 0.28-0.30g of ash. A slight difference was found for contents of 489 mg/ml of K + and 20 mg/ml of NA +. In addition, the phenolic content of the methanolic extracts gives a value of almost 37 mg EAG / g for both cultivars of soy flour. The estimated Reductive Antioxidant Iron (FRAP) potency of soy flour might be related to its polyphenol richness, which is similar to the variety of China. The flour Soya varieties tested contained a significant amount of protein and phenolic compounds with good antioxidant properties.

Keywords: soye beans, soya flour, protein, total polyphenols

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608 In vitro Antifungal Activity of Methanolic Extracts of Eight Various Cultivar of Persian Punica granatum L. against Candida Species

Authors: Shahindokht Bassiri-Jahromi, Mohammad Reza Pourshafie, Farzad Katiraee, Mannan Hajimahmoodi, Ehsan Mostafavi, Malihe Talebi

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Objective: Resistance of Candida species to antifungal agents has potentially serious implications for management of infections. Candida species are now fourth common organisms isolated from hospitalized patients. It is important to increase effective therapy. In the past decade, numerous reports of treatment failures were reported. Prevention and control of these infections will require new antimicrobial agents. Plant-derived antifungal have always been a source of novel therapeutics. The aim of this study was to investigate the antifungal effect of methanolic extract of pomegranate peel and pulp against Candida species. Material and Methods: Eight cultivars of Punica granatum L. were collected from Saveh Agricultural Investigation Center in Iran. Both pomegranate pulp and peel were dried and powdered separately. The dried powders were extracted by using a soxhlet extractor. The antifungal effect of methanolic extract of pomegranate peel and pulp were determined in vitro by minimum inhibitory concentration (MIC) against five standard species of (ATCC 10231), C. parapsilosis (ATCC 22019), C. tropicalis (ATCC 750), C. glabrata (PTCC 5297), and C. kroseii (PTCC 5295). Results: Maximum inhibitions of antifungal effect were attributed to peel extract pomegranate cultivar and Candida species. The most potential antifungal inhibition among 8 different cultivars observed by sour malas, sour white peel, and sour summer extracts respectively, against five Candida strains. The antifungal activity of pulp extracts against Candida species was approximately negative. Conclusion: The use of Punica granatum peel extract has been shown to possess antifungal activities. The phytochemistry and pharmacological actions of Punica granatum peel components suggest a wide range of clinical applications for the treatment and prevention of candidiasis.

Keywords: antifungal activity, Candida species, Punica granatum L., pharmacognosy

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607 Antibacterial and Antioxidant Properties of Total Phenolics from Waste Orange Peels

Authors: Kanika Kalra, Harmeet Kaur, Dinesh Goyal

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Total phenolics were extracted from waste orange peels by solvent extraction and alkali hydrolysis method. The most efficient solvents for extracting phenolic compounds from waste biomass were methanol (60%) > dimethyl sulfoxide > ethanol (60%) > distilled water. The extraction yields were significantly impacted by solvents (ethanol, methanol, and dimethyl sulfoxide) due to varying polarity and concentrations. Extraction of phenolics using 60% methanol yielded the highest phenolics (in terms of gallic acid equivalent (GAE) per gram of biomass) in orange peels. Alkali hydrolyzed extract from orange peels contained 7.58±0.33 mg GAE g⁻¹. By using the solvent extraction technique, it was observed that 60% methanol is comparatively the best-suited solvent for extracting polyphenolic compounds and gave the maximum yield of 4.68 ± 0.47 mg GAE g⁻¹ in orange peel extracts. DPPH radical scavenging activity and reducing the power of orange peel extract were checked, where 60% methanolic extract showed the highest antioxidant activity, 85.50±0.009% for DPPH, and dimethyl sulfoxide (DMSO) extract gave the highest yield of 1.75±0.01% for reducing power ability of the orange peels extract. Characterization of the polyphenolic compounds was done by using Fourier transformation infrared (FTIR) spectroscopy. Solvent and alkali hydrolysed extracts were evaluated for antibacterial activity using the agar well diffusion method against Gram-positive Bacillus subtilis MTCC441 and Gram-negative Escherichia coli MTCC729. Methanolic extract at 300µl concentration showed an inhibition zone of around 16.33±0.47 mm against Bacillus subtilis, whereas, for Escherichia coli, it was comparatively less. Broth-based turbidimetric assay revealed the antibacterial effect of different volumes of orange peel extracts against both organisms.

Keywords: orange peels, total phenolic content, antioxidant, antibacterial

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606 Evalution of Antiurolithiatic Potentials from Cucumis sativus Fruits

Authors: H. J. Pramod, S. Pethkar

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The evaluation of antiurolithiatic potentials from the extracts of Cucumis sativus fruits at different doses and cystone (standard formulation) at a dose of 750 mg/kg were measured for both preventive and curative regimen in wistar rats by adding 0.75% v/v ethylene glycol (EG) to drinking water for 28 days, except normal rats. After the completion of the experimental period, (28th day) urinary parameters like (urine volume, routine urine analysis, levels of calcium, phosphate, oxalate, magnesium, sodium) serum biomarkers like (creatinine, BUN, uric acid, ALP, ALT, AST) kidney homogenate analysis for (levels of calcium, oxalate and phosphate) were analysed. The treated groups shows increased in the urine output significantly compared to the normal. The extract shows significantly decreased in the urinary excretion of the calcium, phosphate, magnesium, sodium and oxalate. The both preventive and curative treatment of extracts showed decrease in the stone forming constituents in the kidneys of urolithiatic rats further the kidneys of all the groups were excised and sectioned for histopathological examination which further claims to posses antiurolithiatic activity.

Keywords: Cucumis sativus, urolithiasis, ethylene glycol, cystone

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605 Formulation and Characterization of Active Edible Films from Cassava Starch for Snacks and Savories

Authors: P. Raajeswari, S. M. Devatha, S. Yuvajanani, U. Rashika

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Edible food packaging are the need of the hour to save life on land and under water by eliminating waste cycle and replacing Single Use Plastics at grass root level as it can be eaten or composted as such. Cassava (Manihot esculenta) selected for making edible films are rich source of starch, and also it exhibit good sheeting propertiesdue to the high amylose: amylopectin content. Cassava starch was extracted by manual method at a laboratory scale and yielded 65 per cent. Edible films were developed by adding food grade plasticizers and water. Glycerol showed good plasticizing property as compared to sorbitol and polylactic acid in both manual (petri dish) and machine (film making machine) production. The thickness of the film is 0.25±0.03 mm. Essential oil and components from peels like pomegranate, orange, pumpkin, onion, and banana brat, and herbs like tulsi and country borage was extracted through the standardized aqueous and alkaline method. In the standardized film, the essential oil and components from selected peel and herbs were added to the casting solution separately and casted the film. It was added to improve the anti-oxidant, anti-microbial and optical properties. By inclusion of extracts, it reduced the bubble formation while casting. FTIR, Water Vapor and Oxygen Transmission Rate (WVTR and OTR), tensile strength, microbial load, shelf life, and degradability of the films were done to analyse the mechanical property of the standardized films. FTIR showed the presence of essential oil. WVTR and OTR of the film was improved after inclusion of essential oil and extracts from 1.312 to 0.811 cm₃/m₂ and 15.12 to 17.81 g/ m₂.d. Inclusion of essential oil from herbs showed better WVTR and OTR than the inclusion of peel extract and standard. Tensile strength and Elongation at break has not changed by essential oil and extracts at 0.86 ± 0.12 mpa and 14 ± 2 at 85 N force. By inclusion of extracts, an optical property of the film enhanced, and it increases the appearance of the packaging material. The films were completely degraded on 84thdays and partially soluble in water. Inclusion of essential oil does not have impact on degradability and solubility. The microbial loads of the active films were decreased from 15 cfu/gm to 7 cfu/gm. The films can be stored at frozen state for 24 days and 48 days at atmospheric temperature when packed with South Indian snacks and savories.

Keywords: active films, cassava starch, plasticizer, characterization

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604 Green Extraction Processes for the Recovery of Polyphenols from Solid Wastes of Olive Oil Industry

Authors: Theodora-Venetia Missirli, Konstantina Kyriakopoulou, Magdalini Krokida

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Olive mill solid waste is an olive oil mill industry by-product with high phenolic, lipid and organic acid concentrations that can be used as a low cost source of natural antioxidants. In this study, extracts of Olea europaea (olive tree) solid olive mill waste (SOMW) were evaluated in terms of their antiradical activity and total phenolic compounds concentrations, such as oleuropein, hydroxytyrosol etc. SOMW samples were subjected to drying prior to extraction as a pretreatment step. Two drying processes, accelerated solar drying (ASD) and air-drying (AD) (at 35, 50, 70°C constant air velocity of 1 m/s), were applied. Subsequently, three different extraction methods were employed to recover extracts from untreated and dried SOMW samples. The methods include the green Microwave Assisted (MAE) and Ultrasound Assisted Extraction (UAE) and the conventional Soxhlet extraction (SE), using water and methanol as solvents. The efficiency and selectivity of the processes were evaluated in terms of extraction yield. The antioxidant activity (AAR) and the total phenolic content (TPC) of the extracts were evaluated using the DPPH assay and the Folin-Ciocalteu method, respectively. The results showed that bioactive content was significantly affected by the extraction technique and the solvent. Specifically, untreated SOMW samples showed higher performance in the yield for all solvents and higher antioxidant potential and phenolic content in the case of water. UAE extraction method showed greater extraction yields than the MAE method for both untreated and dried leaves regardless of the solvent used. The use of ultrasound and microwave assisted extraction in combination with industrially applied drying methods, such as air and solar drying, was feasible and effective for the recovery of bioactive compounds.

Keywords: antioxidant potential, drying treatment, olive mill pomace, microwave assisted extraction, ultrasound assisted extraction

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603 Comparative Production of Secondary Metabolites by Prunus africana (Hook. F.) Kalkman Provenances in Cameroon and Some Associated Endophytic Fungi

Authors: Gloria M. Ntuba-Jua, Afui M. Mih, Eneke E. T. Bechem

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Prunus africana (Hook. F.) Kalkman, commonly known as Pygeum or African cherry belongs to the Rosaceae family. It is a medium to large, evergreen tree with a spreading crown of 10 to 20 m. It is used by the traditional medical practitioners for the treatment of over 45ailments in Cameroon and sub-Sahara Africa. In modern medicine, it is used in the treatment of benign prostrate hyperplasia (BPH), prostate gland hypertrophy (enlarged prostate glands). This is possible because of its ability to produce some secondary metabolites which are believed to have bioactivity against these ailments. The ready international market for the sale of Prunus bark, uncontrolled exploitation, illegal harvesting using inappropriate techniques and poor timing of harvesting have contributed enormously to making the plant endangered. It is known to harbor a large number of endophytic fungi with the potential to produce similar secondary metabolites as the parent plant. Alternative sourcing of medicinal principles through endophytic fungi requires succinct knowledge of the endophytic fungi. This will serve as a conservation measure for Prunus africana by reducing dependence on Prunus bark for such metabolites. This work thus sought to compare the production of some major secondary metabolites produced by P. africana and some of its associated endophytic fungi. The leaves and stem bark of the plant from different provenances were soaked in methanol for 72 hrs to yield the methanolic crude extract. The phytochemical screening of the methanolic crude extracts using different standard procedures revealed the presence of tannins, flavonoids, terpenoids, saponins, phenolics and steroids. Pure cultures of some predominantly isolated endophyte species from the difference Prunus provenances such as Curvularia sp, and Morphospecies P001 were also grown in Potato Dextrose Broth (PDB) for 21 days and later extracted with Methylene dichloride (MDC) solvent after 24hrs to produce crude culture extracts. Qualitative assessment of crude culture extracts showed the presence of tannins, terpenoids, phenolics and steroids particularly β-Sitosterol, (a major bioactive metabolite) as did the plant tissues. Qualitative analysis by thin layer chromatography (TLC) was done to confirm and compare the production of β-Sitosterol (as marker compounds) in the crude extracts of the plant and endophyte. Samples were loaded on TLC silica gel aluminium barked plate (Kieselgel 60 F254, 0.2 mm, Merck) using acetone/hexane, (3.0:7.0) solvent system. They were visualized under an ultra violet lamp (UV254 and UV360). TLC revealed that leaves had a higher concentration of β-sitosterol in terms of band intensity than stem barks from the different provenances. The intensity of β-sitosterol bands in the culture extracts of endophytes was comparable to the plant extracts except for Curvularia sp (very minute) whose band was very faint. The ability of these fungi to make β-sitosterol was confirmed by TLC analysis with the compound having chromatographic properties (retention factor) similar to those of β-sitosterol standard. The ability of these major endophytes to produce secondary metabolites similar to the host has therefore been demonstrated. There is, therefore, the potential of developing the in vitro production system of Prunus secondary metabolites thereby enhancing its conservation.

Keywords: Caneroon, endophytic fungi, Prunus africana, secondary metabolite

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602 Inhibition of Escherichia coli and Salmonella spp. By Traditional Phytomedicines That Are Commonly Used to Treat Gastroenteritis in Zimbabwe

Authors: Constance Chivengwa, Tinashe Mandimutsira, Jephris Gere, Charles Magogo, Irene Chikanza, Jerneja Vidmar, Walter Chingwaru

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The use of traditional methods in the management of diarrhoea has remained a common practice among the indigenous African tribes of Southern Africa. Despite the widespread use of traditional medicines in Zimbabwe, very little research validating the activities of phytomedicines against diarrhoea, as claimed by the Shona people of Zimbabwe, has been reported. This study sought to determine the efficacies of the plants that are frequently used to treat stomach complaints, namely Dicoma anomala, Cassia abbreviata, Lannea edulis and Peltophorum africanum against Escherichia coli (an indicator of faecal contamination of water, and whose strains such as EHEC (O157), ETEC and EPEC, are responsible for a number of outbreaks of diarrhoea) and Salmonella spp. Ethanol and aqueous extracts from these plants were obtained, evaporated, dried and stored. The dried extracts were reconstituted and diluted 10-fold in nutrient broth (from 100 to 0.1 microgram/mL) and tested for inhibition against the bacteria. L. edulis exhibited the best antimicrobial effect (minimum inhibition concentration = 10 microgram/mL for both extracts and microorganisms). Runners up to L. edulis were C. abbreviata (20 microgram/mL for both microorganisms) and P. africanum (20 and 30 microgram/mL respectively). Interestingly, D. anomala, which is widely considered panacea in African medicinal practices, showed low antimicrobial activity (60 and 100 microgram/mL respectively). The high antimicrobial activity of L. edulis can be explained by its content of flavonoids, tannins, alkylphenols (cardonol 7 and cardonol 13) and dihydroalkylhexenones. The antimicrobial activities of C. abbreviata can be linked to its content of anthraquinones and triterpenoids. P. africanum is known to contain benzenoids, flavanols, flavonols, terpenes, xanthone and coumarins. This study therefore demonstrated that, among the plants that are used against diarrhoea in African traditional medicine, L. edulis is a clear winner against E. coli and Salmonella spp. Activity guided extraction is encouraged to establish the complement of compounds that have antimicrobial activities.

Keywords: diarrhoea, Escherichia coli, Salmonella, phytomedicine, MIC, Zimbabwe

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601 Investigation of the Function of Chemotaxonomy of White Tea on the Regulatory Function of Genes in Pathway of Colon Cancer

Authors: Fereydoon Bondarian, Samira Shaygan

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Today, many nutritionists recommend the consumption of plants, fruits, and vegetables to provide the antioxidants needed by the body because the use of plant antioxidants usually causes fewer side effects and better treatment. Natural antioxidants increase the power of plasma antioxidants and reduce the incidence of some diseases, such as cancer. Bad lifestyles and environmental factors play an important role in increasing the incidence of cancer. In this study, different extracts of white teas taken from two types of tea available in Iran (clone 100 and Chinese hybrid) due to the presence of a hydroxyl functional group in their structure to inhibit free radicals and anticancer properties, using 3 aqueous, methanolic and aqueous-methanolic methods were used. The total polyphenolic content was calculated using the Folin-Ciocalcu method, and the percentage of inhibition and trapping of free radicals in each of the extracts was calculated using the DPPH method. With the help of high-performance liquid chromatography, a small amount of each catechin in the tea samples was obtained. Clone 100 white tea was found to be the best sample of tea in terms of all the examined attributes (total polyphenol content, antioxidant properties, and individual amount of each catechin). The results showed that aqueous and aqueous-methanolic extracts of Clone 100 white tea have the highest total polyphenol content with 27.59±0.08 and 36.67±0.54 (equivalent gallic acid per gram dry weight of leaves), respectively. Due to having the highest level of different groups of catechin compounds, these extracts have the highest property of inhibiting and trapping free radicals with 66.61±0.27 and 71.74±0.27% (mg/l) of the extracted sample against ascorbic acid). Using the MTT test, the inhibitory effect of clone 100 white tea extract in inhibiting the growth of HCT-116 colon cancer cells was investigated and the best time and concentration treatments were 500, 150 and 1000 micrograms in 8, 16 and 24 hours, respectively. To investigate gene expression changes, selected genes, including tumorigenic genes, proto-oncogenes, tumor suppressors, and genes involved in apoptosis, were selected and analyzed using the real-time PCR method and in the presence of concentrations obtained for white tea. White tea extract at a concentration of 1000 μg/ml 3 times 16, 8, and 24 hours showed the highest growth inhibition in cancer cells with 53.27, 55.8, and 86.06%. The concentration of 1000 μg/ml aqueous extract of white tea under 24-hour treatment increased the expression of tumor suppressor genes compared to the normal sample.

Keywords: catechin, gene expression, suppressor genes, colon cell line

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600 Formulation and Evaluation of Antioxidant Cream Containing Nepalese Medicinal Plants

Authors: Ajaya Acharya, Prem Narayan Paudel, Rajendra Gyawali

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Due to strong tyrosinase inhibition and antioxidant effects, green tea and Licorice are valuable in cosmetics for the skin. However, data on the addition of essential oils to green tea and Licorice in cream formulation to examine antioxidant activities are limited. The purpose of this study was to develop and assess a phytocosmetic cream’s antioxidant and tyrosinase inhibitory characteristics using crude aqueous extracts of green tea, Licorice, and loaded with essential oils. To load the best concentration on cream formulations, plant aqueous extracts were designed, evaluated, and correlated in terms of total phenolic content (TPC), total flavonoids content (TFC), and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. Moreover, o. tenuiflorum and o. basilicum essential oils were extracted and added to a cream formulation. The spreadability profile, water washability, centrifugation test, and organoleptic characteristics of formulated oil in water cream were all satisfactory. The cream exhibited a non-Newtonian rheological profile and pH range of 6.353 ± 0.065 to 6.467±0.050 over successive 0, 1, 2, and 3 months at normal room temperature. The 50% inhibition concentrations shown by herbal cream were 13.764 ± 0.153 µg/ml, 301.445 ± 1.709 µg/ml and 8.082 ± 0.055 respectively for 2, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity, ferric (Fe³⁺) reducing antioxidant power (FRAP) and 2, 2’-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity, and that of standard ascorbic acid were 6.716 ± 0.077 µg/ml, 171.604 ± 1.551µg/ml and 5.645±0.034µg/ml which showed formulated cream had strong antioxidant characteristics. The formulated herbal cream with a 50% tyrosinase inhibition concentration of 22.254 ± 0.369µg/ml compared to standard Kojic acid 12.535 ± 0.098µg/ml demonstrated a satisfactory tyrosinase inhibition profile for skin whitening property. Herbal cream was reportedly stable in physical and chemical parameters for successive 0, 1, 2, and 3 months at both real and accelerated time study zones, according to obtained stability study results.

Keywords: crude extracts, antioxidant, tyrosinase inhibition, green tea polyphenols

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599 NMR-Based Metabolomic Study of Antimalarial Plant Species Used Traditionally by Vha-Venda People in Limpopo Province, South Africa

Authors: Johanna Bapela, Heino Heyman, Marion Meyer

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Regardless of the significant advances accomplished in reducing the burden of malaria and other tropical diseases in recent years, malaria remains a major cause of mortality in endemic countries. This is especially the case in sub-Saharan Africa where 99% of the estimated global malaria deaths occurs on an annual basis. The emergence of resistant Plasmodium species and the lack of diversified chemotherapeutic agents provide the rationale for bioprospecting for antiplasmodial scaffolds. Crude extracts from twenty indigenous antimalarial plant species were screened for antimalarial activity and then subjected to 1H NMR-based metabolomic analysis. Ten plant extracts exhibited significant in vitro antiplasmodial activity (IC50 ≤ 5 µg/ml). The Principal Component Analysis (PCA) of the acquired 1H NMR spectra could not separate the analyzed plant extracts according to the detected antiplasmodial bioactivity. Application of supervised Orthogonal Projections to Latent Structures–Discriminant Analysis (OPLS-DA) to the 1H NMR profiles resulted in a discrimination pattern that could be correlated to bioactivity. A contribution plot generated from the OPLS-DA scoring plot illustrated the classes of compounds responsible for the observed grouping. Given the preliminary in vitro results, Tabernaemontana elegans Stapf. (Apocynaceae) and Vangueria infausta Burch. subsp. infausta (Rubiaceae) were subjected to further phytochemical investigations. Two indole alkaloids, dregamine and tabernaemontanine possessing antiplasmodial activity were isolated from T. elegans. Two compounds were isolated from V. infausta subsp. infausta and identified as friedelin (IC50 = 3.01 µg/ml) and morindolide (IC50 = 18.5 µg/ml). While these compounds have been previously identified, this is the first account of their occurrence in the genus Vangueria and their antiplasmodial activity. Based on the results of the study, metabolomics can be used to globally identify classes of plant secondary metabolites that are responsible for antiplasmodial activity.

Keywords: ethnopharmacology, Malaria, medicinal plants, metabolomics

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598 Approach to Honey Volatiles' Profiling by Gas Chromatography and Mass Spectrometry

Authors: Igor Jerkovic

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Biodiversity of flora provides many different nectar sources for the bees. Unifloral honeys possess distinctive flavours, mainly derived from their nectar sources (characteristic volatile organic components (VOCs)). Specific or nonspecific VOCs (chemical markers) could be used for unifloral honey characterisation as addition to the melissopalynologycal analysis. The main honey volatiles belong, in general, to three principal categories: terpenes, norisoprenoids, and benzene derivatives. Some of these substances have been described as characteristics of the floral source, and other compounds, like several alcohols, branched aldehydes, and furan derivatives, may be related to the microbial purity of honey processing and storage conditions. Selection of the extraction method for the honey volatiles profiling should consider that heating of the honey produce different artefacts and therefore conventional methods of VOCs isolation (such as hydrodistillation) cannot be applied for the honey. Two-way approach for the isolation of the honey VOCs was applied using headspace solid-phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE). The extracts were analysed by gas chromatography and mass spectrometry (GC-MS). HS-SPME (with the fibers of different polarity such as polydimethylsiloxane/ divinylbenzene (PDMS/DVB) or divinylbenzene/carboxene/ polydimethylsiloxane (DVB/CAR/PDMS)) enabled isolation of high volatile headspace VOCs of the honey samples. Among them, some characteristic or specific compounds can be found such as 3,4-dihydro-3-oxoedulan (in Centaurea cyanus L. honey) or 1H-indole, methyl anthranilate, and cis-jasmone (in Citrus unshiu Marc. honey). USE with different solvents (mainly dichloromethane or the mixture pentane : diethyl ether 1 : 2 v/v) enabled isolation of less volatile and semi-volatile VOCs of the honey samples. Characteristic compounds from C. unshiu honey extracts were caffeine, 1H-indole, 1,3-dihydro-2H-indol-2-one, methyl anthranilate, and phenylacetonitrile. Sometimes, the selection of solvent sequence was useful for more complete profiling such as sequence I: pentane → diethyl ether or sequence II: pentane → pentane/diethyl ether (1:2, v/v) → dichloromethane). The extracts with diethyl ether contained hydroquinone and 4-hydroxybenzoic acid as the major compounds, while (E)-4-(r-1’,t-2’,c-4’-trihydroxy-2’,6’,6’-trimethylcyclo-hexyl)but-3-en-2-one predominated in dichloromethane extracts of Allium ursinum L. honey. With this two-way approach, it was possible to obtain a more detailed insight into the honey volatile and semi-volatile compounds and to minimize the risks of compound discrimination due to their partial extraction that is of significant importance for the complete honey profiling and identification of the chemical biomarkers that can complement the pollen analysis.

Keywords: honey chemical biomarkers, honey volatile compounds profiling, headspace solid-phase microextraction (HS-SPME), ultrasonic solvent extraction (USE)

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597 Rooibos Extract Antioxidants: In vitro Models to Assess Their Bioavailability

Authors: Ntokozo Dambuza, Maryna Van De Venter, Trevor Koekemoer

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Oxidative stress contributes to the pathogenesis of many diseases and consequently antioxidant therapy has attracted much attention as a potential therapeutic strategy. Regardless of the quantities ingested, antioxidants need to reach the diseased tissues at concentrations sufficient to combat oxidative stress. Bioavailability is thus a defining criterion for the therapeutic efficacy of antioxidants. In addition, therapeutic antioxidants must possess biologically relevant characteristics which can target the specific molecular mechanisms responsible for disease related oxidative stress. While many chemical antioxidant assays are available to quantify antioxidant capacity, they relate poorly to the biological environment and provide no information as to the bioavailability. The present comparative study thus aims to characterise green and fermented rooibos extracts, well recognized for their exceptional antioxidant capacity, in terms of antioxidant bioavailability and efficacy in a disease relevant cellular setting. Chinese green tea antioxidant activity was also evaluated. Chemical antioxidant assays (FRAP, DPPH and ORAC) confirmed the potent antioxidant capacity of both green and fermented rooibos, with green rooibos possessing antioxidant activity superior to that of fermented rooibos and Chinese green tea. Bioavailability was assessed using the PAMPA assay and the results indicate that green and fermented rooibos have a permeation coefficient of 5.7 x 10-6 and 6.9 x 10-6 cm/s, respectively. Chinese green tea permeability coefficient was 8.5 x 10-6 cm/s. These values were comparable to those of rifampicin, which is known to have a high permeability across intestinal epithelium with a permeability coefficient of 5 x 10 -6 cm/s. To assess the antioxidant efficacy in a cellular context, U937 and red blood cells were pre-treated with rooibos and Chinese green tea extracts in the presence of a dye DCFH-DA and then exposed to oxidative stress. Green rooibos exhibited highest activity with an IC50 value of 29 μg/ml and 70 μg/ml, when U937 and red blood cells were exposed oxidative stress, respectively. Fermented rooibos and Chinese green tea had IC50 values of 61 μg/ml and 57 μg/ml for U937, respectively, and 221 μg/ml and 405 μg/ml for red blood cells, respectively. These results indicate that fermented and green rooibos extracts were able to permeate the U937 cells and red blood cell membrane and inhibited oxidation of DCFH-DA to a fluorescent DCF within the cells.

Keywords: rooibos, antioxidants, permeability, bioavailability

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596 Study of the Antimicrobial Activity of the Extract of the Eucalyptus camaldulensis stemming from the Algerian Northeast

Authors: Meksem Nabila, Bordjiba Ouahiba, Meraghni Messaouda, Meksem Amara Leila, Djebar Mohhamed Reda

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The problems of protection of the cultures are being more and more important that they interest great number of farmers and scientists because of the excessive use of the organic phytosanitary products of synthesis that causes fatal damages on the environment. To reduce the inconveniences produced by these pesticides, the use of "biopesticides" originated from plants could be an alternative. The aim of this work is the valuation of a botanical species: Eucalyptus camaldulensis from Northeastern Algeria which extracts are supposed to have an antimicrobial activity, similar to pesticides. The extraction of secondary metabolites from the leaves of E. camaldulensis was realized using methanol and water, and measurements of total polyphenols were made by spectrometric method. Determination of the antimicrobial activity of the extracts at issue was realized in vitro on phyto-pathogenic fungal and bacterial stumps. Tests of comparison were included in the essays by using the chemical pesticidal products of synthesis. The obtained results show that the plant contains polyphenols with an efficiency mattering of the order of 22 %. These polyphenols have a strong fungicidal and bactericidal pesticidal activity against various microbial stumps and the values of the zones of inhibition are more important compared with that obtained in the presence of the chemicals of synthesis (fungicide).

Keywords: eucalyptus camaldulensis, biopesticide, polyphenols, antimicrobial activity

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595 Gas Chromatography-Analysis, Antioxidant, Anti-Inflammatory, and Anticancer Activities of Some Extracts and Fractions of Linum usitatissimum

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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594 Preventive Effect of Stem Back Extracts of Coula edulis Baill. against High-Fat / High Sucrose Diet-Induced Insulin Resistance and Oxidative Stress in Rats

Authors: Eric Beyegue, Boris Azantza, Judith Laure Ngondi, Julius E. Oben

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Background: Insulin resistance (IR) and oxidative stress are associated with obesity, diabetes mellitus, and other cardio metabolic disorders. The aim of this study was to investigate the effect of Coula edulis extracts (CEE) on insulin resistance and oxidative stress markers in high-fat/high sucrose diet-induced insulin resistance in rats. Materials and Methods: Thirty male rats were divided into 6 groups of 5 rats each fed, received daily oral administration of CE extracts for 8 weeks as follows: Group 1 or negative control group, fed with standard diet (SD); Group 2 fed with high-fat/high sucrose diet (HFHS) only; Group3 fed with HFHS + CEAq 200; Group 4 fed with HFHS + CEAq 400; Group 5 fed with HFHS + CEEt 200; Group 6 fed with HFHS + CEEt 400. At the end of the experiment (8 weeks), animals were sacrificed plasma lipid profile, glucose, insulin, oxidative marker and digestive enzyme activities were measured. The homeostasis model assessment for insulin resistance (HOMA-IR) was determined. Results: Feedings with HFHS significantly (p < 0.01) induced plasma hyperglycaemia, hyperinsulinaemia, increased triglyceride, total cholesterol, and low-density lipoprotein levels, decreased high-density lipoprotein levels, alterations of α amylase, and glucose-6-phosphatase activities, and oxidative stress. Daily oral administration with CEE for eight weeks after insulin resistance induction had a hypolipidaemic action, antioxidative activities and modulated metabolic markers. Ethanolic extract at the higher dose had the best effect on body weight gain and insulin resistance, whereas aqueous extract showed the better activity on hyperlipidemia. Conclusion: These results suggest that CEAq and CEEt at 400mg/kg are promising complementary supplements that can be used to protect better from metabolic disorders associated with HFHS.

Keywords: Coula edulis Baill, high-fat / high sucrose diet, insulin resistance, oxidative stress

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593 Synergistic Anti-Proliferation Effect of PLK-1 Inhibitor and Livistona Chinensis Fruit Extracts on Lung Adenocarcinoma A549 Cells

Authors: Min-Chien Su, Tzu-Hsuan Hsu, Guan-Xuan Wu, Shyh-Ming Kuo

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Lung cancer is one of the clinically challenging malignant diseases worldwide. For efficient therapeutics in cancer, combination therapy has developed to acquire a better outcome. PLK-1 was one of the major factors affecting cell mitosis in cancer cells, its inhibitor Bi6727 was proven effective in treating several different cancers namely oral cancer, colon cancer and lung cancer. Despite its low toxicity toward normal cells compared to traditional chemotherapy, it is still yet to be evaluated in detail. Livistona Chinensis (LC) is a Chinese herb that used as a traditional prescription to treat lung cancer. Due to the uncertainty of the efficacy of LC, we utilized a water extraction method to extract the Livistona Chinensis and then lyophilized into powder for further study. In this study we investigated the antiproliferation activities of Bi6727 and LC extracts (LCE) on A549 non-small lung cancer cells. The IC50 of Bi6727 and LCE on A549 are 60 nM and 0.8 mg/mL, respectively. The fluorescent staining images shown nucleolus damage in cells treated with Bi6727 and mitochondrial damage after treated with LCE. A549 cells treated with Bi6727 and LCE showed increased expression of Bax, Caspase-3 and Caspase-9 proteins from Western blot assay. LCE also inhibited A549 cells growth keeping cells at G2-M phase from cell cycle assay. Apoptosis assay results showed that LCE induced late apoptosis of A549 cells. JC-1 assay showed that the mitochondria damaged at the LCE concentration of 0.4 mg/mL. In our preliminary anti-proliferation test of combined LCE and Bi-6727 on A549 cells, we found a dramatically decrease in proliferation after treated with LCE first for 24-h and then Bi-6727 for extra 24-h. This was an important finding regarding synergistic anti-proliferation effect of these drugs, However, the usage, the application sequence of LCE and Bi-6727 on A549 cells and their related mechanisms still need to be evaluated. In summary, the drugs exerted anti-proliferation effect on A549 cells independently. We hopefully combine the usage of these two drugs will bring a different and potential outcome in treating lung cancer.

Keywords: anti-proliferation, A549, Livistona Chinensis fruit extracts, PLK-1 inhibitor

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592 Safety Assessment and Prophylactic Efficacy of Moringa stenopetala Leaf Extract Through Mitigation of Oxidative Stress in BV-2 Microglial Cell

Authors: Stephen Adeniyi Adefegha, Vitor Mostardeiro, Vera Maria Morsch, Ademir F. Morel, Ivana Beatrice Manica Da Cruz, Sabrina Somacal Maria Rosa Chitolina Schetinger

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Moringa stenopetala is often consumed as food and used in folkloric medicine for the management of several diseases. Purpose: This study was set up in order to assess the effect of aqueous extract of Moringa stenopetala on cell viability and oxidative stress biomarkers in BV-2 microglial cells. Aqueous extracts of M. stenopetala were prepared, lyophilized and reconstituted in 0.5% dimethylsulphoxide (DMSO). Cells were treated with M. stenopetala extracts (0.1 - 100 µg/ml) for cell viability and nitric oxide (NO) production tests. However, M. stenopetala extract (50 µg/ml) was used in the treatment of cells for the determination of protein carbonyl content and reactive oxygen species (ROS) level. Incubation of BV-2 microglia cell with M. stenopetala extract maintained cell viability, diminished NO and ROS levels, and reduced protein carbonyl contents Chlorogenic acid, rutin, kaempferol and quercetin derivatives were the main phenolic compounds identified in M. stenopetala leaf extract. These phenolic compounds present in M. stenopetala may be responsible for the mitigation of oxidative stress in BV-2 microglial cells.

Keywords: oxidative stress, BV-2 microglial cell, Moringa stenopetala, cell viability, antioxidant

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591 Spray Drying and Physico-Chemical Microbiological Evaluation of Ethanolic Extracts of Propolis

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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590 Q Eqchi Mayan Piper and Cissampelos Species Alter Reporter Genes and Endogenous Genes Expression in Mc-7 Cells

Authors: Sheila M. Wicks, Gail Mahady, Udesh Patel, Joanna Michel, Armando Caceres

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Introduction: The genus piperaceae contains approximately 1000 species of herbs scrubs small trees and hanging vines distributed in both hemispheres. During our ethno medical work in Guatemala of the 27 plant families documented for us e by the Qeqchi Maya for reproductive disorders the most prominent were the Piperaceae (15%) and Menispermiaceae. Our Previous work showed that extracts from form Piper and Cissampelos species bound to both and progesterone and the estrogen receptors. In this work active extracts from Piper aeruginosibaccum Trelease, P auritum, P tuerckheimii and Cissampels tropaeolifolia were tested in functionalized cell based assays including a SEAP reporter gene and by qPCR of ER-responsive gene expression in MCF-7cells. In the reporter gene assay P aeruginosibaccum was estrogenic and enhanced E2 EFFECTS IN MCF-7 CELLS. P. tuerckheimi was not estrogenic alone but significantly enhanced the effects of E2 on SEAP reporter gene expression. Both altered mRNA expression of E2 responsive genes in MCF-7. Methods: this is collaborative project between University of Illinois at Chicago and University of San Carlos Guatemala City. 144 spices of plants were collected in Guatemala of which 57 used to treat a variety of women's reproductive health. The Genus Piperaraceae contains approximately 1000 species of herbs scrubs and small trees. Active extracts of the plants were tested in functionalized in cell-based bioassays including SEAP reporter genes. Results demonstrated altered mRNA expression of E2 responsive genes in MC-7 cells plants were collected in Guatemala of which 57 used. Conclusion of the 5 plants tested all were shown to contain components of binding to estrogenic receptor to a greater or lesser degree. These effects support the use of QEqchi Maya women in Guatemala for reproductive.

Keywords: reporter genes, MC7, guatemala piperaceae, reproductive health

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589 A New Method of Extracting Polyphenols from Honey Using a Biosorbent Compared to the Commercial Resin Amberlite XAD2

Authors: Farid Benkaci-Alia, Abdelhamid Neggada, Sophie Laurentb

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A new extraction method of polyphenols from honey using a biodegradable resin was developed and compared with the common commercial resin amberlite XAD2. For this purpose, three honey samples of Algerian origin were selected for the different physico-chemical and biochemical parameters study. After extraction of the target compounds by both resins, the polyphenol content was determined, the antioxidant activity was tested, and LC-MS analyses were performed for identification and quantification. The results showed that physico-chemical and biochemical parameters meet the norms of the International Honey commission, and the H1 sample seemed to be of high quality. The optimal conditions of extraction by biodegradable resin were a pH of 3, an adsorption dose of 40 g/L, a contact time of 50 min, an extraction temperature of 60°C and no stirring. The regeneration and reuse number of both resins was three cycles. The polyphenol contents demonstrated a higher extraction efficiency of biosorbent than of XAD2, especially in H1. LC-MS analyses allowed for the identification and quantification of fifteen compounds in the different honey samples extracted using both resins and the most abundant compound was 3,4,5-trimethoxybenzoic acid. In addition, the biosorbent extracts showed stronger antioxidant activities than the XAD2 extracts.

Keywords: extraction, polyphénols, biosorbent, resin amberlite, HPLC-MS

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588 Antimycobacterial Activity of Ethanolic Extract of Artemisia absinthium

Authors: T. Hojageldiyev, Y. Bolmammedov, S. Gurbanaliyev

Abstract:

It is known that drugs used in the treatment of tuberculosis show toxic effect to organism especially to liver besides its therapeutic effect. Because of ineffectiveness of drugs used in the treatment regimen of tuberculosis against multidrug resistance (MDR) and extensively drug-resistance (XDR) tuberculosis requires the development of new treatment methods and new, novel drugs. Considering the usage of Artemisia absinthium in traditional medicine in treatment of wounds which suggests its antibacterial activity it seems that, also it may have significant antimycobacterial activity. The objective of present study was to evaluate antibacterial activity of ethanolic extract of A. absinthium against M. tuberculosis. In this study, the effect of ethanolic extract of A. absinthium was tested against tuberculosis and pharmaco-toxicological properties evaluated on laboratory animals. The 20%, 40%, 70% and 96% ethanolic extracts of A. absinthium prepared then its bacteriostatic and bactericidal activities were evaluated by validated methods. Data were analyzed by GraphPad Prism 7.0 at the level P < 0.05. Results showed that ethanolic extracts of A. absinthium show no toxicological properties with having high LD50. All concentrations of extract show high bacteriostatic activity on M. tuberculosis. 96% ethanolic extract has highest bactericidal effect among other concentrations. By conducting further studies, as a result of our study, antimycobacterial drug can be prepared from A. absinthium.

Keywords: Artemisia absinthium, antimycobacterial, ethanolic extract, Mycobacteria tuberculosis

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