Search results for: in vitro antioxidant.
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 334

Search results for: in vitro antioxidant.

274 Ameliorative Effect of Calocybe indica, a Tropical Indian Edible Mushroom on Hyperglycemia Induced Oxidative Stress

Authors: Shanmugasundaram Krishnakumari, Paramasivam Rajeswari, Subramanian Kathiravan

Abstract:

Mushrooms are a group of fleshy macroscopic fungi. They have been valued throughout the world as both edible and medicine. They are highly nutritious with good amount of quality proteins, vitamins and minerals. An edible mushroom, Calocybe indica was selected to validate its nutritional and medicinal properties. Since tissue damage in hyperglycemia has been related to oxidative stress, we evaluated the enzymatic and non-enzymatic antioxidant status in the serum, liver and kidney since they are the target organs in diabetic complications. From the results, increased oxidative stress and decreased antioxidants might be related to the causation of diabetes mellitus. The treatment in the diabetic rats with the Calocybe indica showed an increase in the antioxidant system and decrease in the production of free radicals. The mushrooms which contain antioxidant phytochemicals has potential free radical scavenging capacity and hence can induce the antioxidant system in the body significantly reduces the generated free radicals thereby maintaining the normal levels of the antioxidants

Keywords: Antioxidants, Calocybe indica, diabetes mellitus, edible mushroom, oxidative stress.

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273 Effect of Ginger and L-Carnitine on the Reproductive Performance of Male Rats

Authors: Ismail I. Abo-Ghanema, El-Nasharty M.A., El-Far A. H., Hanan A.Ghonium

Abstract:

In this study, we investigated the effects of ginger and L-carnitine on the reproductive performance of male rats with respect to semen parameters, male sex hormones and the testicular antioxidant system. A total of sixty mature male albino rats were divided into four groups of fifteen rats. The control group received saline, whereas the other three groups received ginger (100 mg kg-1 d- 1.), L-carnitine (150 mg kg-1 d-1.) or a combination of both ginger (100 mg kg-1 d-1.) and L-carnitine (150 mg kg-1 d-1.) via a stomach tube daily for one month. At the end of the treatment period, the rats were sacrificed, and their sperm characteristics (count, motility and viability), antioxidant enzyme factors levels (reduced glutathione, catalase, superoxide dismutase and total antioxidant capacity) and sex hormone levels (testosterone, Follicle stimulating hormone(FSH) and luteinizing hormone (LH) were analysed. Our results showed that the three experimental treatments improved sperm parameters, antioxidant enzyme activity and testosterone hormone levels; the most pronounced positive effects were observed in the group that received a combination of both ginger and L-carnitine. Therefore, the administration of a combination of ginger and L-carnitine may be beneficial for improving male sexual performance.

Keywords: Ginger, L-Carnitine, Spermatogenesis, Rats.

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272 Antidiabetic and Antioxidative Activities of Butyrolactone I from Aspergillus terreus MC751

Authors: Rizna Triana Dewi, Sanro Tachibana, Ahmad Darmawan

Abstract:

The bioassay-guided isolation and purification of an ethyl acetate extract of Aspergillus terreus MC751 led to the characterization of butyrolactone I as an antidiabetic and antioxidant. The antidiabetic activity of butyrolactone I was evaluated by α- glucosidase and α-amylase inhibition assays. Butyrolactone I demonstrated significant concentration-dependent, mixed-type inhibitory activity against yeast α-glucosidase with an IC50 of 54μM. However, the compound exhibited less activity against rat intestinal α-glucosidase and α-amylase. This is the first report on α-glucosidase inhibitory activity of butyrolactone I. The antioxidative activity of butyrolactone I was evaluated based on scavenging effects on 1,1- diphenyl-2-picrylhydrazyl (DPPH) (IC50 =51 μM) and hydrogen peroxide (IC50= 141 μM) radicals as well as a reducing power assay. The results suggest that butyrolactone I is a promising antidiabetic as well as antioxidant and should be considered for clinical trials.

Keywords: Aspergillus terreus MC751, antidiabetic, antioxidant, Butyrolactone I.

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271 Antioxidant Responses to Different Exposure Regimes of Kazakhstan Light Crude Oil in Livers of Male Albino Rats

Authors: K. Mahmoud, T. Shalahmetova, B. Umbayev, Sh. Deraz

Abstract:

Biochemical investigations were carried out to assess the effect of different exposure regimes of Kazakhstan crude oil (KCO) on hepatic antioxidant defense system in albino rats. Contaminants were delivered under two different dosing regimes, with all treatments receiving the same total contaminant load by the end of the exposure period. Rats in regime A injected with KCO once at a dose of 6 ml/kg bw while in regime B injected multiply at a dose of 1.5 ml/kg bw on day 1, 3, 5 and 8. Antioxidant biomarkers were measured in hepatic tissue after 1, 3, 5 and 8 days. Significant induction was observed in serum aminotransferases (ALT, AST) (p<0.01) and hepatic Glutathione-S-transferase (GST) (p<0.05) in the two exposure regimes, with the majority of significant induction occurring in regime A. Superoxide dismutase (SOD) increased 1-d after injection (p<0.01) but the increase was reduced time dependently thereafter while after 8-d induced again (p<0.01). Malondialdehyde (MDA) significantly induced after 3 and 5-d (p<0.05) in regime A while in regime B was not changed significantly (p>0.05) at short time after exposure. However, there was significant increase after 8-d (p<0.01). Histological examination indicates that crude oil induced pathologic changes from inflammatory cells infiltration to hemorrhage and necrosis of hepatocytes. Acute exposure to crude oil adversely affect hepatic cell so human must avoid such exposure.

Keywords: Kazakhstan crude oil, Antioxidant biomarkers, Histological examination, Dose regime, Rats

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270 Optimization of Deglet-Nour Date (Phoenix dactylifera L.) Phenol Extraction Conditions

Authors: Lekbir Adel, Alloui-Lombarkia Ourida, Mekentichi Sihem, Noui Yassine, Baississe Salima

Abstract:

The objective of this study was to optimize the extraction conditions for phenolic compounds, total flavonoids, and antioxidant activity from Deglet-Nour variety. The extraction of active components from natural sources depends on different factors. The knowledge of the effects of different extraction parameters is useful for the optimization of the process, as well for the ability to predict the extraction yield. The effects of extraction variables, namely types of solvent (methanol, ethanol and acetone) and extraction time (1h, 6h, 12h and 24h) on phenolics extraction yield were evaluated. It has been shown that the time of extraction and types of solvent have a statistically significant influence on the extraction of phenolic compounds from Deglet-Nour variety. The optimised conditions yielded values of 80.19 ± 6.37 mg GAE/100 g FW for TPC, 2.34 ± 0.27 mg QE/100 g FW for TFC and 90.20 ± 1.29% for antioxidant activity were methanol solvent and 6 hours of time. According to the results obtained in this study, Deglet-Nour variety can be considered as a natural source of phenolic compounds with good antioxidant capacity.

Keywords: Deglet-Nour variety, Date palm Fruit, Phenolic compounds, Total flavonoids, Antioxidant activity, Extraction, Optimization.

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269 Antioxidant and Aِntimicrobial Properties of Peptides as Bioactive Components in Beef Burger

Authors: F. M. Abu-Salem, M. H. Mahmoud, M. H. El-Kalyoubi, A. Y. Gibriel, A. A. Abou-Arab Arab

Abstract:

Dried soy protein hydrolysate powder was added to the burger in order to enhance the oxidative stability as well as decreases the microbial spoilage. The soybean bioactive compounds (soy protein hydrolysate) as antioxidant and antimicrobial were added at level of 1, 2 and 3 %.Chemical analysis and physical properties were affected by protein hydrolysate addition. The TBA values were significantly affected (P < 0.05) by the storage period and the level of soy protein hydrolysate. All the tested soybean protein hydrolysate additives showed strong antioxidant properties. Samples of soybean protein hydrolysate showed the lowest (P < 0.05) TBA values at each time of storage. The counts of all determined microbiological indicators were significantly (P < 0.05) affected by the addition of the soybean protein hydrolysate. Decreasing trends of different extent were also observed in samples of the treatments for total viable counts, Coliform, Staphylococcus aureus, yeast and molds. Storage period was being significantly (P < 0.05) affected on microbial counts in all samples Staphylococcus aureus were the most sensitive microbe followed by Coliform group of the sample containing protein hydrolysate, while molds and yeast count showed a decreasing trend but not significant (P < 0.05) until the end of the storage period compared with control sample. Sensory attributes were also performed, added protein hydrolysate exhibits beany flavor which was clear about samples of 3% protein hydrolysate.

Keywords: Antioxidant, antimicrobial, isoflavones, bioactive peptide, antioxidant peptides, soybean protein hydrolysate.

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268 Effect of Recombinant Human Follicle Stimulating Hormone on Meiotic Competence of In Vitro Grown Nili Ravi Buffalo Oocytes

Authors: Muhammad Ijaz Khan, Samina Jalali, Beenish Shahid, S. A. Shami, Muhammad Ikramullah

Abstract:

In the present study, the response of Nili Ravi buffalo oocytes to recombinant human follicle stimulating hormone (rhFSH) (Organon) on meiotic maturation in vitro was examined. Oocytes were matured in vitro in medium containing either 0 or 0.05 IU/ ml rhFSH and the stage of nuclear maturation recorded after 24 hours. The percentage of oocytes in the control group undergoing germinal vesicle breakdown (GVBD) observed after 24 hours of culture was 29 % whereas as in rhFSH group the percentage was 10 % were at this stage (P< 0.001).Thus in the presence of rhFSH, a significantly greater number of oocytes had progressed to the more advanced stages of nuclear maturation. Indeed, the maturation of GV (Germinal Vesicle) stage oocytes to the metaphase II (M II) stage after 24 hours was significantly (P< 0.0001) increased by the addition of rhFSH (82 % VS 47 %). The percentage of degenerated oocytes after 24 hours of culture was 24 % in control group, whereas in rhFSH group the percentage was 8 % after 24 hours. Degeneration of the oocytes after 24 hours was not significantly (P = 0. 9361) decreased.

Keywords: Buffalo, in vitro, oocytes, recombinant FSH.

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267 The Effect of Hylocereus polyrhizus and Hylocereus undatus on Physicochemical, Proteolysis, and Antioxidant Activity in Yogurt

Authors: Zainoldin, K.H., Baba, A.S.

Abstract:

Yogurt is a coagulated milk product obtained from the lactic acid fermentation by the action of Lactobacillus bulgaricus and Streptococcus thermophilus. The additions of fruits into milk may enhance the taste and the therapeutical values of milk products. However fruits also may change the fermentation behaviour. In this present study, the changes in physicochemical, the peptide concentration, total phenolics content and the antioxidant potential of yogurt upon the addition of Hylocereus polyrhizus and Hylocereus undatus (white and red dragon fruit) were investigated. Fruits enriched yogurt (10%, 20%, 30% w/w) were prepared and the pH, TTA, syneresis measurement, peptide concentration, total phenolics content and DPPH antioxidant inhibition percentage were determined. Milk fermentation rate was enhanced in red dragon fruit yogurt for all doses (-0.3606 - -0.4126 pH/h) while only white dragon fruit yogurt with 20% and 30% (w/w) composition showed increment in fermentation rate (-0.3471 - -0.3609 pH/h) compared to plain yogurt (-0.3369pH/h). All dragon fruit enriched yogurts generally showed lower pH readings (pH 3.95 - 4.03) compared to plain yogurt (pH 4.05). Both fruit yogurts showed a higher lactic acid percentage (1.14-1.23%) compared to plain yogurt (1.08%). Significantly higher syneresis percentage (57.19 - 70.32%) compared to plain yogurt (52.93%) were seen in all fruit enriched yogurts. The antioxidant activity of plain yogurt (19.16%) was enhanced by the presence of white and red dragon fruit (24.97- 45.74%). All fruit enriched yogurt showed an increment in total phenolic content (36.44 - 64.43mg/ml) compared to plain yogurt (20.25mg/ml). However, the addition of white and red dragon fruit did not enhance the proteolysis of milk during fermentation. Therefore, it could be concluded that the addition of white and red dragon fruit into yogurt enhanced the milk fermentation rate, lactic acid content, syneresis percentage, antioxidant activity, and total phenolics content in yogurt.

Keywords: Antioxidant activity, Hylocereus polyrhizus, Hylocereus undatus, yogurt

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266 Changes of in vitro Cytokine Production induced by δ-Lactams

Authors: Y. Baba hamed, A. Medjdoub, H. Merzouk, M. Narce

Abstract:

The aim of this work was to study the in vitro effects of δ-lactam 1 and its 4-chlorophenyl derivative 2, on the proliferative responses of human lymphocytes and Th1 and Th2 cytokine secretion. The possible protective role of vitamin E on intracellular stress oxidative induced by these compounds was also investigated. Peripheral blood lymphocytes were isolated using differential centrifugation on a density gradient of Histopaque. They were cultured with mitogen concanavalin A, vitamin E (10 μM) and with different concentrations of the compounds 1 and 2 (0.1 to 10 μM). Proliferation (MTT assay), IL-2, INFγ and IL-4 (Elisa kits), intracellular superoxide anion were determined. 1 and 2 were immunostimulant and increased cytokine secretion with a shift away from Th1 response to Th2. These properties were however accompanied by an increase in intracellular oxidative stress. The presence of vitamin E exhibited protective effects by reducing δ- lactam-induced superoxide anion generation in lymphocytes.

Keywords: Cytokines, δ-Lactams, In vitro Lymphocyte Proliferation, Superoxide Anion

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265 Olive Leaves Extract Restored the antioxidant Perturbations in Red Blood Cells Hemolysate in Streptozotocin Induced Diabetic Rats

Authors: Ismail I. Abo Ghanema, Kadry M. Sadek

Abstract:

Oxidative stress and overwhelming free radicals associated with diabetes mellitus are likely to be linked with development of certain complication such as retinopathy, nephropathy and neuropathy. Treatment of diabetic subjects with antioxidant may be of advantage in attenuating these complications. Olive leaf (Oleaeuropaea), has been endowed with many beneficial and health promoting properties mostly linked to its antioxidant activity. This study aimed to evaluate the significance of supplementation of Olive leaves extract (OLE) in reducing oxidative stress, hyperglycemia and hyperlipidemia in Sterptozotocin (STZ)- induced diabetic rats. After induction of diabetes, a significant rise in plasma glucose, lipid profiles except High density lipoproteincholestrol (HDLc), malondialdehyde (MDA) and significant decrease of plasma insulin, HDLc and Plasma reduced glutathione GSH as well as alteration in enzymatic antioxidants was observed in all diabetic animals. During treatment of diabetic rats with 0.5g/kg body weight of Olive leaves extract (OLE) the levels of plasma (MDA) ,(GSH), insulin, lipid profiles along with blood glucose and erythrocyte enzymatic antioxidant enzymes were significantly restored to establish values that were not different from normal control rats. Untreated diabetic rats on the other hand demonstrated persistent alterations in the oxidative stress marker (MDA), blood glucose, insulin, lipid profiles and the antioxidant parameters. These results demonstrate that OLE may be of advantage in inhibiting hyperglycemia, hyperlipidemia and oxidative stress induced by diabetes and suggest that administration of OLE may be helpful in the prevention or at least reduced of diabetic complications associated with oxidative stress.

Keywords: Diabetes mellitus, olive leaves, oxidative stress, red blood cells

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264 Antioxidant Properties and Nutritive Values of Raw and Cooked Pool Barb (Puntius sophore) of Eastern Himalayas

Authors: Ch. Sarojnalini, Wahengbam Sarjubala Devi

Abstract:

Antioxidant properties and nutritive values of raw and cooked Pool barb, Puntius sophore (Hamilton-Buchanan) of Eastern Himalayas, India were determined. Antioxidant activity of the methanol extract of the raw, steamed, fried and curried Pool barb was evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay. In DPPH scavenging assay the IC50 value of the raw, steamed, fried and curried Pool barb was 1.66 micro-gram/ml, 16.09 micro-gram/ml, 8.99 micro-gram/ml, 0.59 micro-gram/ml whereas the IC50 of the reference ascorbic acid was 46.66miro-gram/ml. These results showed that the fish have high antioxidant activity. Protein content was found highest in raw (20.50±0.08%) and lowest in curried (18.66±0.13%). Moisture content in raw, fried and curried was 76.35±0.09, 46.27±0.14 and 57.46±0.24 respectively. Lipid content was recorded 2.46±0.14% in raw and 21.76±0.10% in curried. Ash content varied from 12.57±0.11 to 22.53±0.07%. The total amino acids varied from 36.79±0.02 and 288.43±0.12 mg/100g. Eleven essential mineral elements were found abundant in all the samples. The samples had considerable amount of Fe ranging from 152.17 to 320.39 milli-gram/100gram, Ca 902.06 to 1356.02 milli-gram/100gram, Zn 91.07 to 138.14 milli-gram/100gram, K 193.25 to 261.56 milli-gram/100gram, Mg 225.06 to 229.10 milli-gram/100gram. Ni was not detected in the curried fish. The Mg and K contents were significantly decreased in frying method; however the Fe, Cu, Ca, Co and Mn contents were increased significantly in all the cooked samples. The Mg and Na contents were significantly increased in curried sample and the Cr content was decreased significantly (p<0.05) in all the cooked samples.

Keywords: Antioxidant property, Pool barb, minerals, amino acids, proximate composition, cooking methods.

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263 Antioxidant Properties, Ascorbic Acid and Total Carotenoid Values of Sweet and Hot Red Pepper Paste: A Traditional Food in Turkish Diet

Authors: Kubra Sayin, Derya Arslan

Abstract:

Red pepper (Capsicum annum L.) has long been recognized as a good source of antioxidants, being rich in ascorbic acid and other phytochemicals. In Turkish cuisine red pepper is sometimes consumed raw in salads and baked as a garnish, but its most wide consumption type is red pepper paste. The processing of red pepper into pepper paste includes various thermal treatment steps such as heating and pasteurizing. There are reports demonstrating an enhancement or reduction in antioxidant activity of vegetables after thermal treatment. So this study was conducted to investigate the total phenolic, ascorbic acid and total carotenoids as well as free radical scavenging activity of raw red pepper and various red pepper pastes obtainable on the market. The samples were analyzed for radical-scavenging activity (RSA) and total polyphenol (TP) content using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Folin-Ciocalteu methods, respectively. Total carotenoids and ascorbic acid contents were determined spectrophotometrically. Results suggest that hot pepper paste contained significantly (P<0.05) higher concentrations of TP than sweet pepper paste. However there is no significant (P>0.05) difference in RSA, ascorbic acid and total carotenoids content between sweet and hot red pepper paste products. It is concluded that the red pepper paste, that has a wide range of consumption in Turkish cuisine, presents a good dose of phenolic compounds and antioxidant capacity and it should be regarded as a functional food.

Keywords: Antioxidant properties, Red pepper paste, Total carotenoids, Total phenolic content.

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262 The Effects of Soil Parameters on Efficiency of Essential Oil from Zingiber zerumbet (L.) Smith in Thailand

Authors: Worakrit Worananthakij, Kamonchanok Doungtadum, Nattagan Mingkwan, Supatsorn Chupong

Abstract:

Natural products from herb have been used in different aspects of life as a result of their various biological activities. Generally, plant growth and production of secondary compounds largely depend on environmental conditions. To better understand this correlation, study on biological activity and soil parameter is necessary. This research aims to study the soil parameters which affect the efficiency of the antioxidant activity of essential oils extracted from the Zingiber zerumbet in three areas of Thailand, including Min Buri district, Bangkok province; Muang district, Chiang Mai province and Kaeng Sanam Nang district, Nakhon Ratchasima province. The soil samples in each area were collected and analyzed in the laboratory. The essential oil of Z. zerumbet in each province was extracted and tested for antioxidant activity by hydrodistillation method and DPPH (2,2-diphenyl-1-picrylhydrazyl radical) assay, respectively. The results showed that, the soil parameters such as pH, nitrogen, potassium and phosphorus elements and exchange of cations of soil specimen from Nakhon Ratchasima province were the highest (P<0.05) (6.10 ±0.03, 0.15 ± 0.04 percent of total nitrogen, 16.67 ± 0.46 mg/L, 3.35 ± 0.65 mg/kg and 12.87 ± 0.11 cmol/kg, respectively). In addition, IC50 (Inhibition Concentrtion of antioxidant at 50%) of Z. zerumbet essential oil collected from Nakhon Ratchasima showed the highest value (P<0.05) (1,400 µg/mL). In conclusion, the soil parameters are once important factor for the efficiency of essential oils extract from Z. zerumbet.

Keywords: Antioxidant, essential oil, herb, soil parameter, Zingiber zerumbet.

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261 Statistical Analysis of the Factors that Influence the Properties of Blueberries from Cultivar Bluecrop

Authors: Raquel P. F. Guiné, Susana R. Matos, Daniela V. T. A. Costa, Fernando J. Gonçalves

Abstract:

Because blueberries are worldwide recognized as a good source of beneficial components, their consumption has increased in the past decades, and so have the scientific works about their properties. Hence, this work was undertaken to evaluate the effect of some production and conservation factors on the properties of blueberries from cultivar Bluecrop. The physical and chemical analyses were done according to established methodologies and then all data was treated using software SPSS for assessment of the possible differences among the factors investigated and/or the correlations between the variables at study. The results showed that location of production influenced some of the berries properties (caliber, sugars, antioxidant activity, color and texture) and that the age of the bushes was correlated with moisture, sugars and acidity, as well as lightness. On the other hand, altitude of the farm only was correlated to sugar content. With regards to conservation, it influenced only anthocyanins content and DPPH antioxidant activity. Finally, the type of extract and the order of extraction had a pronounced influence on all the phenolic properties evaluated.

Keywords: Antioxidant activity, blueberry, conservation, geographical origin, phenolic compounds, statistical analysis.

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260 Effect of Fat Percentage and Prebiotic Composition on Proteolysis, ACE-Inhibitory and Antioxidant Activity of Probiotic Yogurt

Authors: Mohammad B. HabibiNajafi, Saeideh Sadat Fatemizadeh, Maryam Tavakoli

Abstract:

In recent years, the consumption of functional foods, including foods containing probiotic bacteria, has come to notice. Milk proteins have been identified as a source of angiotensin-I-converting enzyme )ACE( inhibitory peptides and are currently the best-known class of bioactive peptides. In this study, the effects of adding prebiotic ingredients (inulin and wheat fiber) and fat percentage (0%, 2% and 3.5%) in yogurt containing probiotic Lactobacillus casei on physicochemical properties, degree of proteolysis, antioxidant and ACE-inhibitory activity within 21 days of storage at 5 ± 1 °C were evaluated. The results of statistical analysis showed that the application of prebiotic compounds led to a significant increase in water holding capacity, proteolysis and ACE-inhibitory of samples. The degree of proteolysis in yogurt increases as storage time elapses (P < 0.05) but when proteolysis exceeds a certain threshold, this trend begins to decline. Also, during storage time, water holding capacity reduced initially but increased thereafter. Moreover, based on our findings, the survival of Lactobacillus casei in samples treated with inulin and wheat fiber increased significantly in comparison to the control sample (P < 0.05) whereas the effect of fat percentage on the survival of probiotic bacteria was not significant (P = 0.095). Furthermore, the effect of prebiotic ingredients and the presence of probiotic cultures on the antioxidant activity of samples was significant (P < 0.05).

Keywords: Yogurt, proteolysis, ACE-inhibitory, antioxidant activity.

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259 Effects of Centrifugation, Encapsulation Method and Different Coating Materials on the Total Antioxidant Activity of the Microcapsules of Powdered Cherry Laurels

Authors: B. Cilek Tatar, G. Sumnu, M. Oztop, E. Ayaz

Abstract:

Encapsulation protects sensitive food ingredients against heat, oxygen, moisture and pH until they are released to the system. It can mask the unwanted taste of nutrients that are added to the foods for fortification purposes. Cherry laurels (Prunus laurocerasus) contain phenolic compounds which decrease the proneness to several chronic diseases such as types of cancer and cardiovascular diseases. The objective of this research was to study the effects of centrifugation, different coating materials and homogenization methods on microencapsulation of powders obtained from cherry laurels. In this study, maltodextrin and mixture of maltodextrin:whey protein with a ratio of 1:3 (w/w) were chosen as coating materials. Total solid content of coating materials was kept constant as 10% (w/w). Capsules were obtained from powders of freeze-dried cherry laurels through encapsulation process by silent crusher homogenizer or microfluidization. Freeze-dried cherry laurels were core materials and core to coating ratio was chosen as 1:10 by weight. To homogenize the mixture, high speed homogenizer was used at 4000 rpm for 5 min. Then, silent crusher or microfluidizer was used to complete encapsulation process. The mixtures were treated either by silent crusher for 1 min at 75000 rpm or microfluidizer at 50 MPa for 3 passes. Freeze drying for 48 hours was applied to emulsions to obtain capsules in powder form. After these steps, dry capsules were grounded manually into a fine powder. The microcapsules were analyzed for total antioxidant activity with DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging method. Prior to high speed homogenization, the samples were centrifuged (4000 rpm, 1 min). Centrifugation was found to have positive effect on total antioxidant activity of capsules. Microcapsules treated by microfluidizer were found to have higher total antioxidant activities than those treated by silent crusher. It was found that increasing whey protein concentration in coating material (using maltodextrin:whey protein 1:3 mixture) had positive effect on total antioxidant activity for both silent crusher and microfluidization methods. Therefore, capsules prepared by microfluidization of centrifuged mixtures can be selected as the best conditions for encapsulation of cherry laurel powder by considering their total antioxidant activity. In this study, it was shown that capsules prepared by these methods can be recommended to be incorporated into foods in order to enhance their functionality by increasing antioxidant activity.

Keywords: Antioxidant activity, cherry laurel, microencapsulation, microfluidization.

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258 The Evaluation of Antioxidant and Antimicrobial Activities of Essential Oil and Aqueous, Methanol, Ethanol, Ethyl Acetate and Acetone Extract of Hypericum scabrum

Authors: A. Heshmati, M. Y Alikhani, M. T. Godarzi, M. R. Sadeghimanesh

Abstract:

Herbal essential oil and extracts are a good source of natural antioxidants and antimicrobial compounds. Hypericum is one of the potential sources of these compounds. In this study, the antioxidant and antimicrobial activity of essential oil and aqueous, methanol, ethanol, ethyl acetate and acetone extract of Hypericum scabrum was assessed. Flowers of Hypericum scabrum were collected from the surrounding mountains of Hamadan province and after drying in the shade, the essential oil of the plant was extracted by Clevenger and water, methanol, ethanol, ethyl acetate and acetone extract was obtained by maceration method. Essential oil compounds were identified using the GC-Mass. The Folin-Ciocalteau and aluminum chloride (AlCl3) colorimetric method was used to measure the amount of phenolic acid and flavonoids, respectively. Antioxidant activity was evaluated using DPPH and FRAP. The minimum inhibitory concentration (MIC) and the minimum bacterial/fungicide concentration (MBC/MFC) of essential oil and extracts were evaluated against Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Salmonella typhimurium, Aspergillus flavus and Candida albicans. The essential oil yield of was 0.35%, the lowest and highest extract yield was related to ethyl acetate and water extract. The most component of essential oil was α-Pinene (46.35%). The methanol extracts had the highest phenolic acid (95.65 ± 4.72 µg galic acid equivalent/g dry plant) and flavonoids (25.39 ± 2.73 µg quercetin equivalent/g dry plant). The percentage of DPPH radical inhibition showed positive correlation with concentrations of essential oil or extract. The methanol and ethanol extract had the highest DDPH radical inhibitory. Essential oil and extracts of Hypericum had antimicrobial activity against the microorganisms studied in this research. The MIC and MBC values for essential oils were in the range of 25-25.6 and 25-50 μg/mL, respectively. For the extracts, these values were 1.5625-100 and 3.125-100 μg/mL, respectively. Methanol extracts had the highest antimicrobial activity. Essential oil and extract of Hypericum scabrum, especially methanol extract, have proper antimicrobial and antioxidant activity, and it can be used to control the oxidation and inhibit the growth of pathogenic and spoilage microorganisms. In addition, it can be used as a substitute for synthetic antioxidant and antimicrobial compounds.

Keywords: Antimicrobial, antioxidant, extract, hypericum.

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257 Novel Solid Lipid Nanoparticles for Oral Delivery of Oxyresveratrol: Effect of the Formulation Parameters on the Physicochemical Properties and in vitro Release

Authors: Y. Sangsen, K. Likhitwitayawuid, B. Sritularak, K. Wiwattanawongsa, R. Wiwattanapatapee

Abstract:

Novel solid lipid nanoparticles (SLNs) were developed to improve oral bioavailability of oxyresveratrol (OXY). The SLNs were prepared by a high speed homogenization technique, at an effective speed and time, using Compritol® 888 ATO (5% w/w) as the solid lipid. The appropriate weight proportions (0.3% w/w) of OXY affected the physicochemical properties of blank SLNs. The effects of surfactant types on the properties of the formulations such as particle size and entrapment efficacy were also investigated. Conclusively, Tween 80 combined with soy lecithin was the most appropriate surfactant to stabilize OXY-loaded SLNs. The mean particle size of the optimized formulation was 134.40 ± 0.57 nm. In vitro drug release study, the selected S2 formulation showed a retarded release profile for OXY with no initial burst release compared to OXY suspension in the simulated gastrointestinal fluids. Therefore, these SLNs could provide a suitable system to develop for the oral OXY delivery.

Keywords: Solid lipid nanoparticles, Physicochemical properties, in vitro drug release, Oxyresveratrol.

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256 In vitro Environmental Factors Controlling Root Morphological Traits of Pineapple (Ananas comosus L. Merr)

Authors: S. Mohajer, R. M. Taha, M. Adel

Abstract:

Developing our knowledge of when pineapple roots grow can lead to improved water, fertilizer applications, and more precise culture management. This paper presents current understanding of morphological traits in pineapple roots, highlighting studies using incubation periods and various solid MS media treated with different sucrose concentrations and pH, which directly assess in vitro environmental factors. Rooting parameters had different optimal sucrose concentrations and incubation periods. All shoots failed to root in medium supplemented with sucrose at 5 g/L and no roots formed within the first 45 days in medium enriched with sucrose at 10 g/L. After 75 days, all shoots rooted in medium enriched with 10 and 20 g/L sucrose. Moreover, MS medium supplied with 20 g/L sucrose resulted in the longest and the highest number of roots with 27.3 mm and 4.7, respectively. Root function, such as capacity for P and N uptake, declined rapidly with root length. As a result, the longer the incubation period, the better the rooting responses would be.

Keywords: Environmental factors, in vitro rooting, pineapple, tissue culture.

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255 Effect of Different Media and Mannitol Concentrations on Growth and Development of Vandopsis lissochiloides (Gaudich.) Pfitz. under Slow Growth Conditions

Authors: J. Linjikao, P. Inthima, A. Kongbangkerd

Abstract:

In vitro conservation of orchid germplasm provides an effective technique for ex situ conservation of orchid diversity. In this study, an efficient protocol for in vitro conservation of Vandopsis lissochiloides (Gaudich.) Pfitz. plantlet under slow growth conditions was investigated. Plantlets were cultured on different strength of Vacin and Went medium (½VW and ¼VW) supplemented with different concentrations of mannitol (0, 2, 4, 6 and 8%), sucrose (0 and 3%) and 50 g/L potato extract, 150 mL/L coconut water. The cultures were incubated at 25±2 °C and maintained under 20 µmol/m2s light intensity for 24 weeks without subculture. At the end of preservation period, the plantlets were subcultured to fresh medium for growth recovery. The results found that the highest leaf number per plantlet could be observed on ¼VW medium without adding sucrose and mannitol while the highest root number per plantlet was found on ½VW added with 3% sucrose without adding mannitol after 24 weeks of in vitro storage. The results showed that the maximum number of leaves (5.8 leaves) and roots (5.0 roots) of preserved plantlets were produced on ¼VW medium without adding sucrose and mannitol. Therefore, ¼VW medium without adding sucrose and mannitol was the best minimum growth conditions for medium-term storage of V. lissochiloides plantlets.

Keywords: Preservation, Vandopsis, germplasm, in vitro.

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254 Beijerinckia indica Extracellular Extract Mediated Green Synthesis of Silver Nanoparticles with Antioxidant and Antibacterial Activities against Clinical Pathogens

Authors: Gopalu Karunakaran, Matheswaran Jagathambal, Nguyen Van Minh, Evgeny Kolesnikov, Denis Kuznetsov

Abstract:

This work investigated the use of Beijerinckia indica extracellular extract for the synthesis of silver nanoparticles using AgNO3. The formation of nanoparticles was confirmed by different methods, such as UV-Vis absorption spectroscopy, XRD, FTIR, EDX, and TEM analysis. The formation of silver nanoparticles (AgNPs) was confirmed by the change in color from light yellow to dark brown. The absorbance peak obtained at 430 nm confirmed the presence of silver nanoparticles. The XRD analysis showed the cubic crystalline phase of the synthesized nanoparticles. FTIR revealed the presence of groups that acts as stabilizing and reducing agents for silver nanoparticles formation. The synthesized silver nanoparticles were generally found to be spherical in shape with size ranging from 5 to 20 nm, as evident by TEM analysis. These nanoparticles were found to inhibit pathogenic bacterial strains. This work proved that the bacterial extract is a potential eco-friendly candidate for the synthesis of silver nanoparticles with promising antibacterial and antioxidant properties. 

Keywords: Antioxidant activity, antimicrobial activity, Beijerinckia indica, characterisation, extracellular extracts, silver nanoparticles.

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253 Hypolipidemic and Antioxidant Effects of Black Tea Extract and Quercetin in Atherosclerotic Rats

Authors: Wahyu Widowati, Hana Ratnawati, Tjandrawati Mozefis, Dwiyati Pujimulyani, Yelliantty Yelliantty

Abstract:

Background: Atherosclerosis is the main cause of cardiovascular disease (CVD) with complex and multifactorial process including atherogenic lipoprotein, oxidized low density lipoprotein (LDL), endothelial dysfunction, plaque stability, vascular inflammation, thrombotic and fibrinolytic disorder, exercises and genetic factor Epidemiological studies have shown tea consumption inversely associated with the development and progression of atherosclerosis. The research objectives: to elucidate hypolipidemic, antioxidant effects, as well as ability to improve coronary artery’s histopathologyof black tea extract (BTE) and quercetin in atherosclerotic rats. Methods: The antioxidant activity was determined by using Superoxide Dismutase activity (SOD) of serum and lipid peroxidation product (Malondialdehyde) of plasma and lipid profile including cholesterol total, LDL, triglyceride (TG), High Density Lipoprotein (HDL) of atherosclerotic rats. Inducing atherosclerotic, rats were given cholesterol and cholic acid in feed during ten weeks until rats indicated atherosclerotic symptom with narrowed artery and foamy cells in the artery’s wall. After rats suffered atherosclerotic, the high cholesterol feed and cholic acid were stopped and rats were given BTE 450; 300; 150 mg/kg body weight (BW) daily, quercetin 15; 10; 5 mg/kg BW daily, compared to rats were given vitamin E 60 mg/kg/BW; simvastatin 2.7 mg/kg BW, probucol 30 mg/kg BW daily for 21 days (first treatment) and 42 days (second treatment), negative control (normal feed), positive control (atherosclerotic rats). Results: BTE and quercetin could lower cholesterol total, triglyceride, LDL MDA and increase HDL, SOD were comparable with simvastatin, probucol both for 21 days and 42 days treatment, as well to improve coronary arteries histopathology. Conclusions: BTE andquercetin have hypolipidemic and antioxidant effects, as well as improve coronary arteries histopathology in atherosclerotic rats.

Keywords: Black tea, quercetin, atherosclerosis, antioxidant, hypolipidemic, cardiovascular disease.

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252 Metoprolol Tartrate-Ethylcellulose Tabletted Microparticles: Development of a Validated Invitro In-vivo Correlation

Authors: Fatima Rasool, Mahmood Ahmad, Ghulam Murtaza, Haji M. S. Khan, Shujaat A. Khan, Sonia Khiljee, Muhammad Qamar-Uz-Zaman

Abstract:

This study describes the methodology for the development of a validated in-vitro in-vivo correlation (IVIVC) for metoprolol tartrate modified release dosage forms with distinctive release rate characteristics. Modified release dosage forms were formulated by microencapsulation of metoprolol tartrate into different amounts of ethylcellulose by non-solvent addition technique. Then in-vitro and in-vivo studies were conducted to develop and validate level A IVIVC for metoprolol tartrate. The values of regression co-efficient (R2-values) for IVIVC of T2 and T3 formulations were not significantly (p<0.05) different from 1 while the values of R2 for IVIVC of T1 and Mepressor® were significantly (p<0.05) different from 1. Internal prediction errors of IVIVC, calculated from observed Area under Curve (AUC) and predicted AUC, were less than 10%. This study successfully presents a valid level A IVIVC for metoprolol tartrate modified dosage forms.

Keywords: Metoprolol tartrate, Dissolution, Bioavailability, Validated in-vitro in-vivo correlation.

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251 Screening and Evaluation of in vivo and in vitro Generated Insulin Plant (Vernonia divergens) for Antimicrobial and Anticancer Activities

Authors: Santosh Kumar, Anand Prakash, Kanak Sinha, Anita K Verma

Abstract:

Vernonia divergens Benth., commonly known as “Insulin Plant” (Fam: Asteraceae) is a potent sugar killer. Locally the leaves of the plant, boiled in water are successfully administered to a large number of diabetic patients. The present study evaluates the putative anti-diabetic ingredients, isolated from the in vivo and in vitro grown plantlets of V. divergens for their antimicrobial and anticancer activities. Sterilized explants of nodal segments were cultured on MS (Musashige and Skoog, 1962) medium in presence of different combinations of hormones. Multiple shoots along with bunch of roots were regenerated at 1mg l-1 BAP and 0.5 mg l-1 NAA. Micro-plantlets were separated and sub-cultured on the double strength (2X) of the above combination of hormones leading to increased length of roots and shoots. These plantlets were successfully transferred to soil and survived well in nature. The ethanol extract of plantlets from both in vivo & in vitro sources were prepared in soxhlet extractor and then concentrated to dryness under reduced pressure in rotary evaporator. Thus obtainedconcentrated extracts showed significant inhibitory activity against gram negative bacteria like Escherichia coli and Pseudomonas aeruginosa but no inhibition was found against gram positive bacteria. Further, these ethanol extracts were screened for in vitro percentage cytotoxicity at different time periods (24 h, 48 h and 72 h) of different dilutions. The in vivo plant extract inhibited the growth of EAC mouse cell lines in the range of 65, 66, 78, and 88% at 100, 50, 25 & 12.5μg mL-1 but at 72 h of treatment. In case of the extract of in vitro origin, the inhibition was found against EAC cell lines even at 48h. During spectrophotometric scanning, the extracts exhibited different maxima (ʎ) - four peaks in in vitro extracts as against single in in vivo preparation suggesting the possible change in the nature of ingredients during micropropagation through tissue culture techniques.

Keywords: Anti-cancer, Anti-microbial, EAC mouse cell, Tissue culture, Vernonia divergens.

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250 Functional Lipids and Bioactive Compounds from Oil Rich Indigenous Seeds

Authors: Azza. S. Naik, S. S. Lele

Abstract:

Indian subcontinent has a plethora of traditional medicine systems that provide promising solutions to lifestyle disorders in an 'all natural way'. Spices and oilseeds hold prominence in Indian cuisine hence the focus of the current study was to evaluate the bioactive molecules from Linum usitatissinum (LU), Lepidium sativum (LS), Nigella sativa (NS) and Guizotia abyssinica (GA) seeds. The seeds were characterized for functional lipids like omega-3 fatty acid, antioxidant capacity, phenolic compounds, dietary fiber and anti-nutritional factors. Analysis of the seeds revealed LU and LS to be a rich source of α-linolenic acid (41.85 ± 0.33%, 26.71 ± 0.63%), an omega 3 fatty acid (using GCMS). While studying antioxidant potential NS seeds demonstrated highest antioxidant ability (61.68 ± 0.21 TEAC/ 100 gm DW) due to the presence of phenolics and terpenes as assayed by the Mass spectral analysis. When screened for anti-nutritional factor cyanogenic glycoside, LS seeds showed content as high as 1674 ± 54 mg HCN / kg. GA is a probable good source of a stable vegetable oil (SFA: PUFA 1:2.3). The seeds showed diversified bioactive profile and hence further studies to use different bio molecules in tandem for the development of a possible 'nutraceutical cocktail' have been initiated..

Keywords: antioxidants, bioactives, functional lipids and oilseeds

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249 The Effect of Carboxymethyl Cellulose on the Stability of Emulsions Stabilized by Whey Proteins under Digestion in vitro and in vivo

Authors: D. Leskauskaite, I. Jasutiene, M. Kersiene, E. Malinauskyte, P. Matusevicius

Abstract:

In vitro gastro-duodenal digestion model was used to investigate the changes of emulsions under digestion conditions. Oil in water emulsions stabilized by whey proteins (2%) and stabilized by whey proteins (2%) with addition of carboxymethyl cellulose (0.75%) as gelling agent of continuous phase were prepared at pH7. Both emulsions were destabilized under gastric conditions; however the protective role of carboxymethyl cellulose was indicated by recording delay of fat digestibility of this emulsion. In the presence of carboxymethyl cellulose whey proteins on the interfacial surface of droplets were more resistant to gastric degradation causing limited hydrolysis of fat due to the poor acceptability of lipids for the enzymes. Studies of emulsions using in vivo model supported results from in vitro studies. Lower content of triglycerides in blood serum and higher amount of fecal fat of rats were determined when rats were fed by diet containing emulsion made with whey proteins and carboxymethyl cellulose. 

Keywords: Digestibility, emulsions, lipids, rats.

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248 Effects of Pterocarpus mildbraedii Leaf Extract and Its Fractions on Cadmium and Lead Chloride-Induced Testicular Damage in Male Albino Rats

Authors: R. U. Hamzah, H. L. Muhammad, A. Sayyadi, M. B Busari, R. Garba, M. B. Umar, A. N Abubakar

Abstract:

Lead (Pb) and Cadmium (Cd) are toxic, non-essential transition metals that pose many health risks for both humans and animals. They are environmental toxicants which contribute to testicular damage resulting to infertility problem among male populace worldwide. Chelating agents used for lead and cadmium toxicity are not readily available, toxic, expensive and unable to mop up most of the toxic metals accumulated in various organs. In this study, the effect of crude extract (CE), ethyl acetate fraction (EF) and acetone fraction (AF) of Pterocarpus mildbraedii leaf extract was assessed on cadmium-lead chloride induced testicular damaged in male albino Wistar rats. CE of the leaf was obtained by extracting in absolute methanol which was further subjected to solvent partitioning via vacuum liquid chromatographic (VLC) techniques using ethyl acetate, acetone and 70% methanol. A preliminary phytochemical screening and in vitro antioxidants guided activities on the CE and fractions were determined using standard methods. EF, AF and CE which exhibited significant in vitro activity were subjected to an in vivo study using Wistar rats. In vivo antioxidant markers, male reproductive hormones, testicular enzymes and DNA damage markers were analyzed on the rats’ testes supernatant. AF had the highest quantities of phenols (319.00 mg/g), flavonoids (8.87 mg/g) and tannins (8.87 mg/g) while methanol and EFs were richer in saponins (135.32 µg/g) and alkaloids (38.34 µg/g) respectively. A dose dependent 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP) and lipid peroxidation were observed in all the extract with high antioxidants power in CE and AF. Administration of lead-cadmium chloride solution significantly (p > 0.05) decreases the testicular superoxide dismutase (SOD) activity to 6.82 unit/mg protein, Catalase (CAT) activity to 8.07 of H2O2 consumed/unit/mg protein and Glutathione (GSH) concentration to 31.30 ug/mg protein. There was a concomitant increase in the level of Malondialdehyde (MDA) to a value of 23.70 mmol/mg protein. In addition, lead-cadmium chloride solution significantly (p > 0.05) increases the testicular marker enzymes (Alkaline phosphatase (119.57 u/L), lactate dehydrogenase (357.05 u/L), Acid phosphatase (98.65 u/L)) and DNA damage markers (conjugated dienes (93.39 nmol/mg protein), carbonyl protein (35.39 nmol/mg protein), DNA fragmentation percentage (32.12%)) with lowered testicular hormones (Testosterone (3.1 ng/mL), Follicle stimulating (0.35 IU/mL) and Luteinizing hormones (0.15 IU/mL)) of the animals in negative control group when compared with other treated groups. Treatment with Pterocarpus mildbraedii leaf extract reverts the observed changes with the best activities found in the CE and AFs in a dose dependent manner. Pterocarpus mildbraedii leaf extract ameliorated the lead/cadmium induced testicular damage in male albino rats. The restoration of the aforementioned parameters by some of the extract dosages were comparable to the standard drug with higher activities in the crude and AF. Therefore, Pterocarpus mildbraedii leaf extract can be explored further for the management of lead/cadmium induced toxicity.

Keywords: Cadmium, lead, Pterocarpus mildbraedii, testicular damage.

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247 The Effect of Buckwheat (Fagopyrum esculentum Moench) Groats Addition to the Lard Diet on Antioxidant Parameters of Plasma and Selected Tissues in Wistar Rats

Authors: Chlopicka Joanna, Barton Henryk, Kryczyk Jadwiga, Francik Renata

Abstract:

Recent studies demonstrated that high-fat diet increases oxidative stress in plasma and in a variety of tissues. Many researchers have been looking for natural products, which can reverse the effect of high fat diet. Recently, buckwheat is becoming common ingredient in functional food because of it properties. In study on buckwheat, it is known that, this plant plays roles as anti-oxidative, anti-inflammatory and anti-hypertensive. Nevertheless still little is known about buckwheat groats. The aim of this study was to investigate the effects of addition of buckwheat groats to the fat diet (30% lard), on some antioxidant and oxidant stress parameters in plasma and selected tissues in Wistar rats. The experiment was carried out with three months old male Wistar rats ca. 250g of body weight fed for 5 weeks with either a high-fat (30% of lard) diet or control diet, with or without addition of buckwheat groats. In plasma biochemistry and the activities of the antioxidant enzymes were measured selected tissues: glutathione peroxidase (GPX), catalase (CAT) and the levels of total and reduced glutathione (GSH), free thiol groups (pSH), antioxidant potential of plasma (FRAP) and oxidant stress indices - proteins carbonyl groups (CO) and malonyldialdehyde concentration (MDA). Activity of catalase (CAT) in plasma of rats was significantly increased in buckwheat groats groups and activity of GPx3 in plasma of rats was decreased in buckwheat groups as compared to control group. The reduced glutathione (GSH) in plasma of rats was significantly increased and protein CO was significantly decreased in buckwheat groups as compared to controls. The lowered concentration of GSH was found in serum of rats fed buckwheat groats addition but it accompanied in 7-fold increase in reduced-to-oxidized glutatione ratio, significant increase in HDL and decrease in nonHDL concentration. Conclusions: Buckwheat groats indicate a beneficial effect in inhibiting protein and lipid peroxidation in rats and improved lipid profile. These results suggest that buckwheat groats exert a significant antioxidant potential and may be used as normal food constituent to ameliorate the oxidant-induced damage in organism. 

Keywords: Antioxidant, buckwheat, high-fat diet, rats.

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246 Comparison of Classical and Ultrasound-Assisted Extractions of Hyphaene thebaica Fruit and Evaluation of Its Extract as Antibacterial Activity in Reducing Severity of Erwinia carotovora

Authors: Hanan Moawad, Naglaa M. Abd EL-Rahman

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Erwinia carotovora var. carotovora is the main cause of soft rot in potatoes. Hyphaene thebaica was studied for biocontrol of E. carotovora which inhibited growth of E. carotovora on solid medium, a comparative study of classical and ultrasound-assisted extractions of Hyphaene thebaica fruit. The use of ultrasound decreased significant the total time of treatment and increase the total amount of crude extract. The crude extract was subjected to determine the in vitro, by a bioassay technique revealed that the treatment of paper disks with ultrasound extraction of Hyphaene thebaica reduced the growth of pathogen and produced inhibition zones up to 38mm in diameter. The antioxidant activity of ultrasound-ethanolic extract of Doum fruits (Hyphaene thebaica) was determined. Data obtained showed that the extract contains the secondary metabolites such as Tannins, Saponin, Flavonoids, Phenols, Steroids, Terpenoids, Glycosides and Alkaloids.

Keywords: Ultrasound, classical extract, Biological control, Erwinia carotovora, Hyphaene thebaica.

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245 Micropropagation and in vitro Conservation via Slow Growth Techniques of Prunus webbii (Spach) Vierh: An Endangered Plant Species in Albania

Authors: Valbona Sota, Efigjeni Kongjika

Abstract:

Wild almond is a woody species, which is difficult to propagate either generatively by seed or by vegetative methods (grafting or cuttings) and also considered as Endangered (EN) in Albania based on IUCN criteria. As a wild relative of cultivated fruit trees, this species represents a source of genetic variability and can be very important in breeding programs and cultivation. For this reason, it would be of interest to use an effective method of in vitro mid-term conservation, which involves strategies to slow plant growth through physicochemical alterations of in vitro growth conditions. Multiplication of wild almond was carried out using zygotic embryos, as primary explants, with the purpose to develop a successful propagation protocol. Results showed that zygotic embryos can proliferate through direct or indirect organogenesis. During subculture, stage was obtained a great number of new plantlets identical to mother plants derived from the zygotic embryos. All in vitro plantlets obtained from subcultures underwent in vitro conservation by minimal growth in low temperature (4ºC) and darkness. The efficiency of this technique was evaluated for 3, 6, and 10 months of conservation period. Maintenance in these conditions reduced micro cuttings growth. Survival and regeneration rates for each period were evaluated and resulted that the maximal time of conservation without subculture on 4ºC was 10 months, but survival and regeneration rates were significantly reduced, specifically 15.6% and 7.6%. An optimal period of conservation in these conditions can be considered the 5-6 months storage, which can lead to 60-50% of survival and regeneration rates. This protocol may be beneficial for mass propagation, mid-term conservation, and for genetic manipulation of wild almond.

Keywords: Micropropagation, minimal growth, storage, wild almond.

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