Search results for: short-chain fatty acids
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1054

Search results for: short-chain fatty acids

154 Mentha piperita Formulations in Natural Deep Eutectic Solvents: Phenolic Profile and Biological Activity

Authors: Tatjana Jurić, Bojana Blagojević, Denis Uka, Ružica Ždero Pavlović, Boris M. Popović

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Natural deep eutectic solvents (NADES) represent a class of modern systems that have been developed as a green alternative to toxic organic solvents, which are commonly used as extraction media. It has been considered that hydrogen bonding is the main interaction leading to the formation of NADES. The aim of this study was phytochemical characterization and determination of the antioxidant and antibacterial activity of Mentha piperita leaf extracts obtained by six choline chloride-based NADES. NADES were prepared by mixing choline chloride with different hydrogen bond donors in 1:1 molar ratio following the addition of 30% (w/w) water. The mixtures were then heated (60 °C) and stirred (650 rpm) until the clear homogenous liquids were obtained. The Mentha piperita extracts were prepared by mixing 75 mg of peppermint leaves with 1 mL of NADES following by the heating and stirring (60 °C, 650 rpm) within 30 min. The content of six phenolics in extracts was determined using HPLC-PDA. The dominant compounds presented in peppermint leaves - rosmarinic acid and luteolin 7-O-glucoside, were extracted by NADES at a similar level as 70% ethanol. The microdilution method was applied to test the antibacterial activity of extracts. Compared with 70% ethanol, all NADES systems showed higher antibacterial activity towards Pseudomonas aeruginosa (Gram -), Staphylococcus aureus (Gram +), Escherichia coli (Gram -), and Salmonella enterica (Gram -), especially NADES containing organic acids. The majority of NADES extracts showed a better ability to neutralize DPPH radical than conventional solvent and similar ability to reduce Fe3+ to Fe2+ ions in FRAP assay. The obtained results introduce NADES systems as the novel, sustainable, and low-cost solvents with a variety of applications.

Keywords: antibacterial activity, antioxidant activity, green extraction, natural deep eutectic solvents, polyphenols

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153 Safety Evaluation of Post-Consumer Recycled PET Materials in Chilean Industry by Overall Migration Tests

Authors: Evelyn Ilabaca, Ximena Valenzuela, Alejandra Torres, María José Galotto, Abel Guarda

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One of the biggest problems in food packaging industry, especially with the plastic materials, is the fact that these materials are usually obtained from non-renewable resources and also remain as waste after its use, causing environmental issues. This is an international concern and particular attention is given to reduction, reuse and recycling strategies for decreasing the waste from plastic packaging industry. In general, polyethylenes represent most plastic waste and recycling process of post-consumer polyethylene terephthalate (PCR-PET) has been studied. US Food and Drug Administration (FDA), European Food Safety Authority (EFSA) and Southern Common Market (MERCOSUR) have generated different legislative documents to control the use of PCR-PET in the production of plastic packaging intended direct food contact in order to ensure the capacity of recycling process to remove possible contaminants that can migrate into food. Consequently, it is necessary to demonstrate by challenge test that the recycling process is able to remove specific contaminants, obtaining a safe recycled plastic to human health. These documents establish that the concentration limit for substitute contaminants in PET is 220 ppb (ug/kg) and the specific migration limit is 10 ppb (ug/kg) for each contaminant, in addition to assure the sensorial characteristics of food are not affected. Moreover, under the Commission Regulation (EU) N°10/2011 on plastic materials and articles intended to come into contact with food, it is established that overall migration limit is 10 mg of substances per 1 dm2 of surface area of the plastic material. Thus, the aim of this work is to determine the safety of PCR-PET-containing food packaging materials in Chile by measuring their overall migration, and their comparison with the established limits at international level. This information will serve as a basis to provide a regulation to control and regulate the use of recycled plastic materials in the manufacture of plastic packaging intended to be in direct contact with food. The methodology used involves a procedure according to EN-1186:2002 with some modifications. The food simulants used were ethanol 10 % (v/v) and acetic acid 3 % (v/v) as aqueous food simulants, and ethanol 95 % (v/v) and isooctane as substitutes of fatty food simulants. In this study, preliminary results showed that Chilean food packaging plastics with different PCR-PET percentages agree with the European Legislation for food aqueous character.

Keywords: contaminants, polyethylene terephthalate, plastic food packaging, recycling

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152 Induction of Callus and Expression of Compounds in Capsicum Frutescens Supplemented with of 2, 4-D

Authors: Jamilah Syafawati Yaacob, Muhammad Aiman Ramli

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Cili padi or Capsicum frutescens is one of capsicum species from nightshade family, Solanaceae. It is famous in Malaysia and is widely used as a food ingredient. Capsicum frutescens also possess vast medicinal properties. The objectives of this study are to determine the most optimum 2,4-D hormone concentration for callus induction from stem explants C. frutescens and the effects of different 2,4-D concentrations on expression of compounds from C. frutescens. Seeds were cultured on MS media without hormones (MS basal media) to yield aseptic seedlings of this species, which were then used to supply explant source for subsequent tissue culture experiments. Stem explants were excised from aseptic seedlings and cultured on MS media supplemented with various concentrations (0.1, 0.3 and 0.5 mg/L) of 2,4-D to induce formation of callus. Fresh weight, dry weight and callus growth percentage in all samples were recorded. The highest mean of dry weight was observed in MS media supplemented with 0.5 mg/L 2,4-D, where 0.4499 ± 0.106 g of callus was produced. The highest percentage of callus growth (16.4%) was also observed in cultures supplemented with 0.5 mg/L 2,4-D. The callus samples were also subjected to HPLC-MS to evaluate the effect of hormone concentration on expression of bio active compounds in different samples. Results showed that caffeoylferuloylquinic acids were present in all samples, but was most abundant in callus cells supplemented with 0.3 & 0.5 mg/L 2,4-D. Interestingly, there was an unknown compound observed to be highly expressed in callus cells supplemented with 0.1 mg/L 2,4-D, but its presence was less significant in callus cells supplemented with 0.3 and 0.5 mg/L 2,4-D. Furthermore, there was also a compound identified as octadecadienoic acid, which was uniquely expressed in callus supplemented with 0.5 mg/L 2,4-D, but absent in callus cells supplemented with 0.1 and 0.3 mg/L 2,4-D. The results obtained in this study indicated that plant growth regulators played a role in expression of secondary metabolites in plants. The increase or decrease of these growth regulators may have triggered a change in the secondary metabolite biosynthesis pathways, thus causing differential expression of compounds in this plant.

Keywords: callus, in vitro, secondary metabolite, 2, 4-Dichlorophenoxyacetic acid

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151 Investigation of Fumaric Acid Radiolysis Using Gamma Irradiation

Authors: Wafa Jahouach-Rabai, Khouloud Ouerghi, Zohra Azzouz-Berriche, Faouzi Hosni

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Widely used organic products in the pharmaceutical industry have been detected in environmental systems, essentially carboxylic acids. In this purpose, the degradation efficiency of these contaminants was evaluated using an advanced oxidation process (AOP), namely ionization process as an alternative to conventional water treatment technologies. This process permitted the generation of radical reactions to directly degrade organic pollutants in wastewater. In fact, gamma irradiation of aqueous solutions produces several reactive radicals, essentially hydroxyl radical (OH), to destroy recalcitrant pollutants. Different concentrations of aqueous solutions of Fumaric acid (FA) were considered in this study (0.1-1 mmol/L), which were treated by irradiation doses from 1 to 15 kGy with 6.1 kGy/h rate by ionizing system in pilot scale (⁶⁰Co irradiator). Variations of main parameters influencing degradation efficiency versus absorbed doses were released in the aim to optimize total mineralization of considered pollutants. Preliminary degradation pathway until complete mineralization into CO₂ has been suggested based on detection of residual degradation derivatives using different techniques, namely high performance liquid chromatography (HPLC) and electron paramagnetic resonance spectroscopy (EPR). Results revealed total destruction of treated compound, which improve the efficiency of this process in water remediation. We investigated the reactivity of hydroxyl radicals generated by irradiation on dicarboxylic acid (FA) in aqueous solutions, leading to its degradation into other smaller molecules. In fact, gamma irradiation of FA leads to the formation of hydroxylated intermediates such as hydroxycarbonyl radical which were identified by EPR spectroscopy. Finally, pilot plant irradiation facilities improved the applicability of radiation technology on large scale.

Keywords: AOP, radiolysis, fumaric acid, gamma irradiation, hydroxyl radical, EPR, HPLC

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150 Production, Characterisation and Assessment of Biomixture Fuels for Compression Ignition Engine Application

Authors: K. Masera, A. K. Hossain

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Hardly any neat biodiesel satisfies the European EN14214 standard for compression ignition engine application. To satisfy the EN14214 standard, various additives are doped into biodiesel; however, biodiesel additives might cause other problems such as increase in the particular emission and increased specific fuel consumption. In addition, the additives could be expensive. Considering the increasing level of greenhouse gas GHG emissions and fossil fuel depletion, it is forecasted that the use of biodiesel will be higher in the near future. Hence, the negative aspects of the biodiesel additives will likely to gain much more importance and need to be replaced with better solutions. This study aims to satisfy the European standard EN14214 by blending the biodiesels derived from sustainable feedstocks. Waste Cooking Oil (WCO) and Animal Fat Oil (AFO) are two sustainable feedstocks in the EU (including the UK) for producing biodiesels. In the first stage of the study, these oils were transesterified separately and neat biodiesels (W100 & A100) were produced. Secondly, the biodiesels were blended together in various ratios: 80% WCO biodiesel and 20% AFO biodiesel (W80A20), 60% WCO biodiesel and 40% AFO biodiesel (W60A40), 50% WCO biodiesel and 50% AFO biodiesel (W50A50), 30% WCO biodiesel and 70% AFO biodiesel (W30A70), 10% WCO biodiesel and 90% AFO biodiesel (W10A90). The prepared samples were analysed using Thermo Scientific Trace 1300 Gas Chromatograph and ISQ LT Mass Spectrometer (GC-MS). The GS-MS analysis gave Fatty Acid Methyl Ester (FAME) breakdowns of the fuel samples. It was found that total saturation degree of the samples was linearly increasing (from 15% for W100 to 54% for A100) as the percentage of the AFO biodiesel was increased. Furthermore, it was found that WCO biodiesel was mainly (82%) composed of polyunsaturated FAMEs. Cetane numbers, iodine numbers, calorific values, lower heating values and the densities (at 15 oC) of the samples were estimated by using the mass percentages data of the FAMEs. Besides, kinematic viscosities (at 40 °C and 20 °C), densities (at 15 °C), heating values and flash point temperatures of the biomixture samples were measured in the lab. It was found that estimated and measured characterisation results were comparable. The current study concluded that biomixture fuel samples W60A40 and W50A50 were perfectly satisfying the European EN 14214 norms without any need of additives. Investigation on engine performance, exhaust emission and combustion characteristics will be conducted to assess the full feasibility of the proposed biomixture fuels.

Keywords: biodiesel, blending, characterisation, CI engine

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149 The Effects of Arginine, Glutamine and Threonine Supplementation in the Starting Phase on Subsequent Performance of Male Broile

Authors: Jalal Fazli Amiri, Mohammad Hossein Shahir, Mohammad Hossein Nemati, Afshin Heidarinia

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The current study was performed to investigate the effects of arginine, threonine, and glutamine supplementation in excess of requirements in the starter period (17 days) on performance, intestinal morphology, and immune response of broilers. Four hundred and sixteen male day-old chicks were assigned in a 2×2×2 factorial arrangement to a completely randomized design with four replicates (13 birds per replicate ). Treatments were: a control group that received the basal diet, basal diet plus 1% glutamine, basal diet plus 0.2% threonine, basal diet plus 0.75 % arginine, and combination of these three amino acids (glutamine+arginine, glutamine+threonine, arginine+threonine and arginine+ glutamine+threonine). The effect of glutamine supplementation on feed intake was significant in week 4 (p < 0.05), week 6 (p < 0.001), and total feed intake (p < 0.05) and caused declined feed intake. No significant differences of glutamine addition were observed on intestinal morphology (villi height, crypt depth, villi height to crypt depth ratio, villi width). Threonine supplementation caused increased weight gain in week 2 (p < 0.001) and 3 and a decrease of total feed intake (p < 0.05). Duodenum and jejunum villi height, crypt depth, villi height to crypt depth ratio, villi width were not affected. The effect of arginine supplementation was the increase of breast percentage (p < 0.05) and a decrease of jejunum villi high (p < 0.05) and Jejunum crypt depth (p < 0.05). Supplementation of arginine, threonine, and glutamine had no significant effects on blood titer of antibodies against Newcastle disease, infectious bronchitis, avian influenza. Overall, it seems that the supplementation of arginine, threonine, and glutamine in excess of requirements in the starter period had no effect on performance in subsequent periods and intestinal morphology.

Keywords: intestinal morphology, immunity, broiler chickens, glutamine, arginine, threonine

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148 A Review: Role of Chromium in Broiler

Authors: Naveed Zahra, Zahid Kamran, Shakeel Ahmad

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Heat stress is one of the most important environmental stressors challenging poultry production worldwide. The detrimental effect of heat stress results in reduction in the productive performance of poultry with high incidences of mortality. Researchers have made efforts to prevent such damage to poultry production through dietary manipulation. Supplementation with Chromium (Cr) might have some positive effects on some aspect of blood parameters and broilers performance. Chromium (Cr) the element whose trivalent Cr (III) organic state is present in trace amounts in animal feed and water is found to be a key element in evading heat stress and thus cutting down the heavy expenditure on air conditioning in broiler sheds. Chromium, along with other essential minerals is lost due to increased excretion during heat stress and thus its inclusion in broiler diet is kind of mandatory in areas of hot climate. Chromium picolinate in broiler diet has shown a hike in growth rate including muscle gain with body fat reduction under environmental stress. Fat reduction is probably linked to the ability of chromium to increase the sensitivity of the insulin receptors on tissues and thus the uptake of sugar from blood increases which decreases the amount of glucose to be converted to amino acids and stored in adipose tissue as triglycerides. Organic chromium has also shown to increase lymphocyte proliferation rate and antioxidant levels. So, the immune competency, muscle gain and fat reduction along with evasion of heat stress are good enough signs that indicate the fruitful inclusion of dietary chromium for broiler. This promising element may bring the much needed break in the local poultry industry. The task is now to set the exact dose of the element in the diet that would be useful enough and still not toxic to broiler. In conclusion there is a growing body of evidence which suggest that chromium may be an essential trace element for livestock and poultry. The nutritional requirement for chromium may vary with different species and physiological state within a species.

Keywords: broiler, chromium, heat stress, performance

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147 Phylogenetic Analysis Based On the Internal Transcribed Spacer-2 (ITS2) Sequences of Diadegma semiclausum (Hymenoptera: Ichneumonidae) Populations Reveals Significant Adaptive Evolution

Authors: Ebraheem Al-Jouri, Youssef Abu-Ahmad, Ramasamy Srinivasan

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The parasitoid, Diadegma semiclausum (Hymenoptera: Ichneumonidae) is one of the most effective exotic parasitoids of diamondback moth (DBM), Plutella xylostella in the lowland areas of Homs, Syria. Molecular evolution studies are useful tools to shed light on the molecular bases of insect geographical spread and adaptation to new hosts and environment and for designing better control strategies. In this study, molecular evolution analysis was performed based on the 42 nuclear internal transcribed spacer-2 (ITS2) sequences representing the D. semiclausum and eight other Diadegma spp. from Syria and worldwide. Possible recombination events were identified by RDP4 program. Four potential recombinants of the American D. insulare and D. fenestrale (Jeju) were detected. After detecting and removing recombinant sequences, the ratio of non-synonymous (dN) to synonymous (dS) substitutions per site (dN/dS=ɷ) has been used to identify codon positions involved in adaptive processes. Bayesian techniques were applied to detect selective pressures at a codon level by using five different approaches including: fixed effects likelihood (FEL), internal fixed effects likelihood (IFEL), random effects method (REL), mixed effects model of evolution (MEME) and Program analysis of maximum liklehood (PAML). Among the 40 positively selected amino acids (aa) that differed significantly between clades of Diadegma species, three aa under positive selection were only identified in D. semiclausum. Additionally, all D. semiclausum branches tree were highly found under episodic diversifying selection (EDS) at p≤0.05. Our study provide evidence that both recombination and positive selection have contributed to the molecular diversity of Diadegma spp. and highlights the significant contribution of D. semiclausum in adaptive evolution and influence the fitness in the DBM parasitoid.

Keywords: diadegma sp, DBM, ITS2, phylogeny, recombination, dN/dS, evolution, positive selection

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146 BiFormerDTA: Structural Embedding of Protein in Drug Target Affinity Prediction Using BiFormer

Authors: Leila Baghaarabani, Parvin Razzaghi, Mennatolla Magdy Mostafa, Ahmad Albaqsami, Al Warith Al Rushaidi, Masoud Al Rawahi

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Predicting the interaction between drugs and their molecular targets is pivotal for advancing drug development processes. Due to the time and cost limitations, computational approaches have emerged as an effective approach to drug-target interaction (DTI) prediction. Most of the introduced computational based approaches utilize the drug molecule and protein sequence as input. This study does not only utilize these inputs, it also introduces a protein representation developed using a masked protein language model. In this representation, for every individual amino acid residue within the protein sequence, there exists a corresponding probability distribution that indicates the likelihood of each amino acid being present at that particular position. Then, the similarity between each pair of amino-acids is computed to create similarity matrix. To encode the knowledge of the similarity matrix, Bi-Level Routing Attention (BiFormer) is utilized, which combines aspects of transformer-based models with protein sequence analysis and represents a significant advancement in the field of drug-protein interaction prediction. BiFormer has the ability to pinpoint the most effective regions of the protein sequence that are responsible for facilitating interactions between the protein and drugs, thereby enhancing the understanding of these critical interactions. Thus, it appears promising in its ability to capture the local structural relationship of the proteins by enhancing the understanding of how it contributes to drug protein interactions, thereby facilitating more accurate predictions. To evaluate the proposed method, it was tested on two widely recognized datasets: Davis and KIBA. A comprehensive series of experiments was conducted to illustrate its effectiveness in comparison to cuttingedge techniques.

Keywords: BiFormer, transformer, protein language processing, self-attention mechanism, binding affinity, drug target interaction, similarity matrix, protein masked representation, protein language model

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145 Potential Use of Cnidoscolus Chayamansa Leaf from Mexico as High-Quality Protein Source

Authors: Diana Karina Baigts Allende, Mariana Gonzalez Diaz, Luis Antonio Chel Guerrero, Mukthar Sandoval Peraza

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Poverty and food insecurity are still incident problems in the developing countries, where population´s diet is based on cereals which are lack in protein content. Nevertheless, during last years the use of native plants has been studied as an alternative source of protein in order to improve the nutritional intake. Chaya crop also called Spinach tree, is a prehispanic plant native from Central America and South of Mexico (Mayan culture), which has been especially valued due to its high nutritional content particularly protein and some medicinal properties. The aim of this work was to study the effect of protein isolation processing from Chaya leaf harvest in Yucatan, Mexico on its structure quality in order: i) to valorize the Chaya crop and ii) to produce low-cost and high-quality protein. Chaya leaf was extruded, clarified and recovered using: a) acid precipitation by decreasing the pH value until reach the isoelectric point (3.5) and b) thermal coagulation, by heating the protein solution at 80 °C during 30 min. Solubilized protein was re-dissolved in water and spray dried. The presence of Fraction I protein, known as RuBisCO (Rubilose-1,5-biphosfate carboxylase/oxygenase) was confirmed by gel electrophoresis (SDS-PAGE) where molecular weight bands of 55 KDa and 12 KDa were observed. The infrared spectrum showed changes in protein structure due to the isolation method. The use of high temperatures (thermal coagulation) highly decreased protein solubility in comparison to isoelectric precipitated protein, the nutritional properties according to amino acid profile was also disturbed, showing minor amounts of overall essential amino acids from 435.9 to 367.8 mg/g. Chaya protein isolate obtained by acid precipitation showed higher protein quality according to essential amino acid score compared to FAO recommendations, which could represent an important sustainable source of protein for human consumption.

Keywords: chaya leaf, nutritional properties, protein isolate, protein structure

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144 Evaluation of Genetic Fidelity and Phytochemical Profiling of Micropropagated Plants of Cephalantheropsis obcordata: An Endangered Medicinal Orchid

Authors: Gargi Prasad, Ashiho A. Mao, Deepu Vijayan, S. Mandal

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The main objective of the present study was to optimize and develop an efficient protocol for in vitro propagation of a medicinally important orchid Cephalantheropsis obcordata (Lindl.) Ormerod along with genetic stability analysis of regenerated plants. This plant has been traditionally used in Chinese folk medicine and the decoction of whole plant is known to possess anticancer activity. Nodal segments used as explants were inoculated on Murashige and Skoog (MS) medium supplemented with various concentrations of isopentenyl adenine (2iP). The rooted plants were successfully acclimatized in the greenhouse with 100% survival rate. Inter-simple sequence repeats (ISSR) markers were used to assess the genetic fidelity of in vitro raised plants and the mother plant. It was revealed that monomorphic bands showing the absence of polymorphism in all in vitro raised plantlets analyzed, confirming the genetic uniformity among the regenerants. Phytochemical analysis was done to compare the antioxidant activities and HPLC fingerprinting assay of 80% aqueous ethanol extract of the leaves and stem of in vitro and in vivo grown C. obcordata. The extracts of the plants were examined for their antioxidant activities by using free radical 1, 1-diphenyl-2-picryl hydrazyl (DPPH) scavenging method, 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging ability, reducing power capacity, estimation of total phenolic content, flavonoid content and flavonol content. A simplified method for the detection of ascorbic acid, phenolic acids and flavonoids content was also developed by using reversed phase high-performance liquid chromatography (HPLC). This is the first report on the micropropagation, genetic integrity study and quantitative phytochemical analysis of in vitro regenerated plants of C. obcordata.

Keywords: Cephalantheropsis obcordata, genetic fidelity, ISSR markers, HPLC

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143 Desulfurization of Crude Oil Using Bacteria

Authors: Namratha Pai, K. Vasantharaj, K. Haribabu

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Our Team is developing an innovative cost effective biological technique to desulfurize crude oil. ’Sulphur’ is found to be present in crude oil samples from .05% - 13.95% and its elimination by industrial methods is expensive currently. Materials required :- Alicyclobacillus acidoterrestrius, potato dextrose agar, oxygen, Pyragallol and inert gas(nitrogen). Method adapted and proposed:- 1) Growth of bacteria studied, energy needs. 2) Compatibility with crude-oil. 3) Reaction rate of bacteria studied and optimized. 4) Reaction development by computer simulation. 5) Simulated work tested by building the reactor. The method being developed requires the use of bacteria Alicyclobacillus acidoterrestrius - an acidothermophilic heterotrophic, soil dwelling aerobic, Sulfur bacteria. The bacteria are fed to crude oil in a unique manner. Its coated onto potato dextrose agar beads, cultured for 24 hours (growth time coincides with time when it begins reacting) and fed into the reactor. The beads are to be replenished with O2 by passing them through a jacket around the reactor which has O2 supply. The O2 can’t be supplied directly as crude oil is inflammable, hence the process. Beads are made to move around based on the concept of fluidized bed reactor. By controlling the velocity of inert gas pumped , the beads are made to settle down when exhausted of O2. It is recycled through the jacket where O2 is re-fed and beads which were inside the ring substitute the exhausted ones. Crude-oil is maintained between 1 atm-270 M Pa pressure and 45°C treated with tartaric acid (Ph reason for bacteria growth) for optimum output. Bacteria being of oxidising type react with Sulphur in crude-oil and liberate out SO4^2- and no gas. SO4^2- is absorbed into H2O. NaOH is fed once reaction is complete and beads separated. Crude-oil is thus separated of SO4^2-, thereby Sulphur, tartaric acid and other acids which are separated out. Bio-corrosion is taken care of by internal wall painting (phenolepoxy paints). Earlier methods used included use of Pseudomonas and Rhodococcus species. They were found to be inefficient, time and energy consuming and reduce the fuel value as they fed on skeleton.

Keywords: alicyclobacillus acidoterrestrius, potato dextrose agar, fluidized bed reactor principle, reaction time for bacteria, compatibility with crude oil

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142 Identification and Quantification of Sesquiterpene Lactones of Sagebrush (Artemisia tridentate) and Its Chemical Modification

Authors: Rosemary Anibogwu, Kavita Sharma, Karl De Jesus

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Sagebrush is an abundant and naturally occurring plant in the Intermountain West region of the United States. The plant contains an array of bioactive compounds such as flavonoids, terpenoids, sterols, and phenolic acids. It is important to identify and characterize these compounds because Native Americans use sagebrush as herbal medicine. These compounds are also utilized for preventing infection in wounds, treating headaches and colds, and possess antitumor properties. This research is an exploratory study on the sesquiterpene present in the leaves of sagebrush. The leaf foliage was extracted with 100 % chloroform and 100 % methanol. The percentage yield for the crude was considerably higher in chloroform. The Thin Layer Chromatography (TLC) analysis of the crude extracted unveiled a brown band at Rf = 0.25 and a dark brown band at Rf = 0.74, along with three unknown faint bands the 254 nm UV lamp. Furthermore, the two distinct brown (Achillin) and dark brown band (Hydroxyachillin) in TLC were further utilized in the isolation of pure compounds with column chromatography. The structures of Achillin and Hydroxyachillin were elucidated based on extensive spectroscopic analysis, including TLC, High-Performance Liquid Chromatography (HPLC), 1D- and 2D-Nuclear Magnetic Resonance (NMR), and Mass Spectroscopy (MS). The antioxidant activities of crude extract and three pure compounds were evaluated in terms of their peroxyl radical scavenging by Ferric Reducing Ability of Plasma (FRAP) and 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) methods. The crude extract showed the antioxidant activity of 18.99 ± 0.51 µmol TEg -1 FW for FRAP and 11.59 ± 0.38 µmol TEg -1 FW for DPPH. The activities of Achillin, Hydroxyachillin, and Quercetagetin trimethyl ether were 13.03, 15.90 and 14.02 µmol TEg -1 FW respectively for the FRAP assay. The three purified compounds have been submitted to the National Cancer Institute 60 cancer cell line for further study.

Keywords: HPLC, nuclear magnetic resonance spectroscopy, sagebrush, sesquiterpene lactones

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141 Phenolic Composition and Antioxidant Activity of Sorbus L. Fruits and Leaves

Authors: Raudone Lina, Raudonis Raimondas, Gaivelyte Kristina, Pukalskas Audrius, Janulis Valdimaras, Viskelis Pranas

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Sorbus L. species are widely distributed in the Northern hemisphere and have been used for medicinal purposes in various traditional medicine systems and as food ingredients. Various Sorbus L. raw materials, fruits, leaves, inflorescences, barks, possess diuretic, anti-inflammatory, hypoglycemic, anti-diarrheal and vasoprotective activities. Phenolics, to whom main pharmacological activities are attributed, are compounds of interest due to their notable antioxidant activity. The aim of this study was to determine the antioxidant profiles of fruits and leaves of selected Sorbus L. species (S. anglica, S. aria f. latifolia, S. arranensis, S. aucuparia, S. austriaca, S. caucasica, S. commixta, S. discolor, S. gracilis, S. hostii, S. semi-incisa, S. tianschanica) and to identify the phenolic compounds with potent contribution to antioxidant activity. Twenty two constituents were identified in Sorbus L. species using ultra high performance liquid chromatography coupled to quadruple and time-of-flight mass spectrometers (UPLC–QTOF–MS). Reducing activity of individual constituents was determined using high performance liquid chromatography (HPLC) coupled to post-column FRAP assay. Signicantly greatest trolox equivalent values corresponding up to 45% of contribution to antioxidant activity were assessed for neochlorogenic and chlorogenic acids, which were determined as markers of antioxidant activity in samples of leaves and fruits. Characteristic patterns of antioxidant profiles obtained using HPLC post-column FRAP assay significantly depend on specific Sorbus L. species and raw materials and are suitable for equivalency research of Sorbus L. fruits and leaves. Selecting species and target plant organs with richest phenolic composition and strongly expressed antioxidant power is the first step in further research of standardized extracts.

Keywords: FRAP, antioxidant, phenolic, Sorbus L., chlorogenic acid, neochlorogenic acid

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140 Increased Expression Levels of Soluble Epoxide Hydrolase in Obese and Its Modulation by Physical Exercise

Authors: Abdelkrim Khadir, Sina Kavalakatt, Preethi Cherian, Ali Tiss

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Soluble epoxide hydrolase (sEH) is an emerging therapeutic target in several chronic states that have inflammation as a common underlying cause such as immunometabolic diseases. Indeed, sEH is known to play a pro-inflammatory role by metabolizing anti-inflammatory, epoxyeicosatrienoic acids (EETs) to pro-inflammatory diols. Recently, it was shown sEH to be linked to diet and microbiota interaction in rat models of obesity. Nevertheless, the functional contribution of sEH and its anti-inflammatory substrates EETs in obesity remain poorly understood. In the current study, we compared the expression pattern of sEH between lean and obese nondiabetic human subjects using subcutaneous adipose tissue (SAT) and peripheral blood mononuclear cells (PBMCs). Using RT-PCR, western blot and immunofluorescence confocal microscopy, we show here that the level of sEH mRNA and protein to be significantly increased in obese subjects with concomitant increase in endoplasmic reticulum (ER) stress components (GRP78 and ATF6α) and inflammatory markers (TNF-α, IL-6) when compared to lean controls. The observation that sEH was overexpressed in obese subjects’ prompt us to investigate whether physical exercise could reduce its expression. In this study, we report here 3-months supervised physical exercise significantly attenuated the expression of sEH in both the SAT and PBMCs, with a parallel decrease in the expression of ER stress markers along with attenuated inflammatory response. On the other hand, homocysteine, a sulfur containing amino acid deriving from the essential amino acid methionine was shown to be directly associated with insulin resistance. When 3T3-L1 preadipocytes cells were treated with homocysteine our results show increased sEH levels along with ER stress markers. Collectively, our data suggest that sEH upregulation is strongly linked to ER stress in adiposity and that physical exercise modulates its expression. This gives further evidence that exercise might be useful as a strategy for managing obesity and preventing its associated complications.

Keywords: obesity, adipose tissue, epoxide hydrolase, ER stress

Procedia PDF Downloads 139
139 Humins: From Industrial By-Product to High Value Polymers

Authors: Pierluigi Tosi, Ed de Jong, Gerard van Klink, Luc Vincent, Alice Mija

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During the last decades renewable and low-cost resources have attracted increasingly interest. Carbohydrates can be derived by lignocellulosic biomasses, which is an attractive option since they represent the most abundant carbon source available in nature. Carbohydrates can be converted in a plethora of industrially relevant compounds, such as 5-hydroxymethylfurfural (HMF) and levulinic acid (LA), within acid catalyzed dehydration of sugars with mineral acids. Unfortunately, these acid catalyzed conversions suffer of the unavoidable formation of highly viscous heterogeneous poly-disperse carbon based materials known as humins. This black colored low value by-product is made by a complex mixture of macromolecules built by covalent random condensations of the several compounds present during the acid catalyzed conversion. Humins molecular structure is still under investigation but seems based on furanic rings network linked by aliphatic chains and decorated by several reactive moieties (ketones, aldehydes, hydroxyls, …). Despite decades of research, currently there is no way to avoid humins formation. The key parameter for enhance the economic viability of carbohydrate conversion processes is, therefore, increasing the economic value of the humins by-product. Herein are presented new humins based polymeric materials that can be prepared starting from the raw by-product by thermal treatment, without any step of purification or pretreatment. Humins foams can be produced with the control of reaction key parameters, obtaining polymeric porous materials with designed porosity, density, thermal and electrical conductivity, chemical and electrical stability, carbon amount and mechanical properties. Physico chemical properties can be enhanced by modifications on the starting raw material or adding different species during the polymerization. A comparisons on the properties of different compositions will be presented, along with tested applications. The authors gratefully acknowledge the European Community for financial support through Marie-Curie H2020-MSCA-ITN-2015 "HUGS" Project.

Keywords: by-product, humins, polymers, valorization

Procedia PDF Downloads 143
138 Exploring Emerging Viruses From a Protected Reserve

Authors: Nemat Sokhandan Bashir

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Threats from viruses to agricultural crops could be even larger than the losses caused by the other pathogens because, in many cases, the viral infection is latent but crucial from an epidemic point of view. Wild vegetation can be a source of many viruses that eventually find their destiny in crop plants. Although often asymptomatic in wild plants due to adaptation, they can potentially cause serious losses in crops. Therefore, exploring viruses in wild vegetation is very important. Recently, omics have been quite useful for exploring plant viruses from various plant sources, especially wild vegetation. For instance, we have discovered viruses such as Ambrossia asymptomatic virus I (AAV-1) through the application of metagenomics from Oklahoma Prairie Reserve. Accordingly, extracts from randomly-sampled plants are subjected to high speed and ultracentrifugation to separated virus-like particles (VLP), then nucleic acids in the form of DNA or RNA are extracted from such VLPs by treatment with phenol—chloroform and subsequent precipitation by ethanol. The nucleic acid preparations are separately treated with RNAse or DNAse in order to determine the genome component of VLPs. In the case of RNAs, the complementary cDNAs are synthesized before submitting to DNA sequencing. However, for VLPs with DNA contents, the procedure would be relatively straightforward without making cDNA. Because the length of the nucleic acid content of VPLs can be different, various strategies are employed to achieve sequencing. Techniques similar to so-called "chromosome walking" may be used to achieve sequences of long segments. When the nucleotide sequence data were obtained, they were subjected to BLAST analysis to determine the most related previously reported virus sequences. In one case, we determined that the novel virus was AAV-l because the sequence comparison and analysis revealed that the reads were the closest to the Indian citrus ringspot virus (ICRSV). AAV—l had an RNA genome with 7408 nucleotides in length and contained six open reading frames (ORFs). Based on phylogenies inferred from the replicase and coat protein ORFs of the virus, it was placed in the genus Mandarivirus.

Keywords: wild, plant, novel, metagenomics

Procedia PDF Downloads 80
137 Impact of Non-Starch Polysaccharides on Sensorial Characteristics and Textural Properties of Bread

Authors: Farhan Saeed, Imran Pasha, Faqir M. Anjum, Muhammad U. Arshad

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Introduction: Cereals especially wheat is one example in this respite as it contains several nutrients and phytochemicals. In this regard, presences of non-starch polysaccharides are of significance value e.g. arabinoxylans (AX) and arabinogalactans (AG). These ingredients possess several functional and nutritional properties and in this project, efforts were directed to extract AX and AG from different spring wheat varieties of Pakistan and subsequent utilization in cereal based baked products. Methodology: In the present study, effort was made to characterize eight different spring wheats e.g. Lasani-08, FSD-08, Mairaj-08, Shafaq-06, Sehar-06, Bhakkar-02, Uqab-2000 and Inqalab-91 with special reference to non-starch polysaccharides (arabinoxylans and arabinogalactans) extraction followed by their utilization in baked products. Major Findings of Study: Results showed that the arabinoxylans and arabinogalactans content in whole wheat flour of different wheat varieties ranged from 2.93 to 4.68% and 0.47 to 0.93%, respectively while in bran, they ranged from 11.71 to 18.38% and 1.07-4.43%, respectively. Phenolic compounds i.e. ferulic acid, p-coumaric acids were 1.12 and 19.6mg/100g, respectively. Owing to presence of these phenolic compounds, it has persuasive antioxidant potential. Arabinoxylan has negative impact on gluten quality as reduced gluten strength was observed while significant results were obtained for rheological characteristic. Moreover, adding Arabinoxylan and arabinogalactan in bread formulation resulted in significant increase in volume and texture of the final product. In addition, the hardness of bread lessened considerably due to the increase in the concentration of arabinoxylan and arabinogalactan. Additionally, fracturability of bread improved as the both non-starch polysaccharides levels increased. The highest gumminess value was given to Shafaq-06 with increasing trend from control to 0.5% arabinoxylan. Whilst with the addition of arabinogalactan, the highest bread gumminess value (155.74 ± 6.1, 156.32 ± 7.9) was also observed in Shafaq-06. Concluding Statement: Conclusively, it may be inferred that non-starch polysaccharides hold potential to be extracted and utilized in cereal based products for best quality and value addition.

Keywords: non-starch polysaccharides, arabinoxylan, arabinogalactan, bread

Procedia PDF Downloads 223
136 Isolation and Characterization of the First Known Inhibitor Cystine Knot Peptide in Sea Anemone: Inhibitory Activity on Acid-Sensing Ion Channels

Authors: Armando A. Rodríguez, Emilio Salceda, Anoland Garateix, André J. Zaharenko, Steve Peigneur, Omar López, Tirso Pons, Michael Richardson, Maylín Díaz, Yasnay Hernández, Ludger Ständker, Jan Tytgat, Enrique Soto

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Acid-sensing ion channels are cation (Na+) channels activated by a pH drop. These proteins belong to the ENaC/degenerin superfamily of sodium channels. ASICs are involved in sensory perception, synaptic plasticity, learning, memory formation, cell migration and proliferation, nociception, and neurodegenerative disorders, among other processes; therefore those molecules that specifically target these channels are of growing pharmacological and biomedical interest. Sea anemones produce a large variety of ion channels peptide toxins; however, those acting on ligand-gated ion channels, such as Glu-gated, Ach-gated ion channels, and acid-sensing ion channels (ASICs), remain barely explored. The peptide PhcrTx1 is the first compound characterized from the sea anemone Phymanthus crucifer, and it constitutes a novel ASIC inhibitor. This peptide was purified by chromatographic techniques and pharmacologically characterized on acid-sensing ion channels of mammalian neurons using patch-clamp techniques. PhcrTx1 inhibited ASIC currents with an IC50 of 100 nM. Edman degradation yielded a sequence of 32 amino acids residues, with a molecular mass of 3477 Da by MALDI-TOF. No similarity to known sea anemone peptides was found in protein databases. The computational analysis of Cys-pattern and secondary structure arrangement suggested that this is a structurally ICK (Inhibitor Cystine Knot)-type peptide, a scaffold that had not been found in sea anemones but in other venomous organisms. These results show that PhcrTx1 represents the first member of a new structural group of sea anemones toxins acting on ASICs. Also, this peptide constitutes a novel template for the development of drugs against pathologies related to ASICs function.

Keywords: animal toxin, inhibitor cystine knot, ion channel, sea anemone

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135 The Effects of Nano Zerovalent Iron (nZVI) and Magnesium Oxide Nanoparticles on Methane Production during Anaerobic Digestion of Waste Activated Sludge

Authors: Passkorn Khanthongthip, John T. Novak

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Many studies have been reported that the nZVI and MgO NPs were often found in waste activated sludge (WAS). However, little is known about the impact of those NPs on WAS stabilization. The aims of this study were to investigate the effects of both NPs on WAS anaerobic digestion for methane production and to examine the change of metanogenic population under those different environments using qPCR. Four dosages (2, 50, 100, and 200 mg/g-TSS) of MgO NPs were added to four different bottles containing WAS to investigate the impact of MgO NPs on methane production during WAS anaerobic digestion. The effects of nZVI on methane production during WAS anaerobic digestion were also conducted in another four bottles using the same methods described above except that the MgO NPs were replaced by nZVI. A bottle of WAS anaerobic digestion without nanoparticles addition was also operated to serve as a control. It was found that the relative amounts, compared to the control system, of methane production in each WAS anaerobic digestion bottle adding 2, 50, 100, 200 mg/gTSS MgO NPs were 98, 62, 28, and 14 %, respectively. This suggests that higher MgO NPs resulted in lower methane production. The data of batch test for the effects of corresponding released Mg2+ indicated that 50 mg/gTSS MgO NPs or higher could inhibit methane production at least 25%. Moreover, the volatile fatty acid (VFA) concentration was 328, 384, 928, 3,684, and 7,848 mg/L for the control and four WAS anaerobic digestion bottles with 2, 50, 100, 200 mg/gTSS MgO NPs addition, respectively. Higher VFA concentration could reduce pH and subsequently decrease methanogen growth, resulting in lower methane production. The relative numbers of total gene copies of methanogens analyzed from samples taken from WAS anaerobic digestion bottles were approximately 99, 68, 38, and 24 % of control for the addition of 2, 50, 100, and 200 mg/gTSS, respectively. Obviously, the more MgO NPs appeared in sludge anaerobic digestion system, the less methanogens remained. In contrast, the relative amount of methane production found in another four WAS anaerobic digestion bottles adding 2, 50, 100, and 200 mg/gTSS nZVI were 102, 128, 112, and 104 % of the control, respectively. The measurement of methanogenic population indicated that the relative content of methanogen gene copies were 101, 132, 120, and 112 % of those found in control, respectively. Additionally, the cumulative VFA was 320, 234, 308, and 330 mg/L, respectively. This reveals that nZVI addition could assist to increase methanogenic population. Higher amount of methanogen accelerated VFA degradation for greater methane production, resulting in lower VFA accumulation in digesters. Moreover, the data for effects of corresponding released Fe2+ conducted by batch tests suggest that the addition of approximately 50 mg/gTSS nZVI increased methane production by 20%. In conclusion, the presence of MgO NPs appeared to diminish the methane production during WAS anaerobic digestion. Higher MgO NPs dosages resulted in more inhibition on methane production. In contrast, nZVI addition promoted the amount of methanogenic population which facilitated methane production.

Keywords: magnesium oxide nanoparticles, methane production, methanogenic population, nano zerovalent iron

Procedia PDF Downloads 295
134 An Alternative Semi-Defined Larval Diet for Rearing of Sand Fly Species Phlebotomus argentipes in Laboratory

Authors: Faizan Hassan, Seema Kumari, V. P. Singh, Pradeep Das, Diwakar Singh Dinesh

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Phlebotomus argentipes is an established vector for Visceral Leishmaniasis in Indian subcontinent. Laboratory colonization of Sand flies is imperative in research on vectors, which requires a proper diet for their larvae and adult growth that ultimately affects their survival and fecundity. In most of the laboratories, adult Sand flies are reared on rabbit blood feeding/artificial blood feeding and their larvae on fine grinded rabbit faeces as a sole source of food. Rabbit faeces are unhygienic, difficult to handle, high mites infestation as well as owing to bad odour which creates menacing to human users ranging from respiratory problems to eye infection and most importantly it does not full fill all the nutrients required for proper growth and development. It is generally observed that the adult emergence is very low in comparison to egg hatched, which may be due to insufficient food nutrients provided to growing larvae. To check the role of food nutrients on larvae survival and adult emergence, a high protein rich artificial diet for sand fly larvae were used in this study. The composition of artificial diet to be tested includes fine grinded (9 gm each) Rice, Pea nuts & Soyabean balls. These three food ingredients are rich source of all essential amino acids along with carbohydrate and minerals which is essential for proper metabolism and growth. In this study artificial food was found significantly more effective for larval development and adult emergence than rabbit faeces alone (P value >0.05). The weight of individual larvae was also found higher in test pots than the control. This study suggest that protein plays an important role in insect larvae development and adding carbohydrate will also enhances the fecundity of insects larvae.

Keywords: artificial food, nutrients, Phlebotomus argentipes, sand fly

Procedia PDF Downloads 305
133 Preparation and Characterization of Supported Metal Nanocrystal Using Simple Heating Method for Renewable Diesel Synthesis from Nyamplung Oil (Calophyllum inophyllum Oil)

Authors: Aida Safiera, Andika Dwi Rubyantoro, Muhammad Bagus Prakasa

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Indonesia’s needs of diesel oil each year are increasing and getting urge. However, that problems are not supported by the amount of oil production that still low and also influenced by the fact of oil reserve is reduced. Because of that, the government prefers to import from other countries than fulfill the needs of diesel. To anticipate that problem, development of fuel based on renewable diesel is started. Renewable diesel is renewable alternative fuel that is hydrocarbon derivative from decarbonylation of non-edible oil. Indonesia is rich with natural resources, including nyamplung oil (Calophyllum inophyllum oil) and zeolite. Nyamplung oil (Calophyllum inophyllum oil) has many stearic acids which are useful on renewable diesel synthesis meanwhile zeolite is cheap. Zeolite is many used on high temperature reaction and cracking process on oil industry. Zeolite also has advantages which are a high crystallization, surface area and pores. In this research, the main focus that becomes our attention is on preparation and characterization of metal nanocrystal. Active site that used in this research is Nickel Molybdenum (NiMo). The advantage of nanocrystal with nano scale is having larger surface area. The synthesis of metal nanocrystal will be done with conventional preparation modification method that is called simple heating. Simple heating method is a metal nanocrystal synthesis method using continuous media which is polymer liquid. This method is a simple method and produces a small particles size in a short time. Influence of metal nanocrystal growth on this method is the heating profile. On the synthesis of nanocrystal, the manipulated variables are temperature and calcination time. Results to achieve from this research are diameter size on nano scale (< 100 nm) and uniform size without any agglomeration. Besides that, the conversion of synthesis of renewable diesel is high and has an equal specification with petroleum diesel. Catalyst activities are tested by FT-IR and GC-TCD on decarbonylation process with a pressure 15 bar and temperature 375 °C. The highest conversion from this reaction is 35% with selectivity around 43%.

Keywords: renewable diesel, simple heating, metal nanocrystal, NiMo, zeolite

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132 Production and Purification of Monosaccharides by Hydrolysis of Sugar Cane Bagasse in an Ionic Liquid Medium

Authors: T. R. Bandara, H. Jaelani, G. J. Griffin

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The conversion of lignocellulosic waste materials, such as sugar cane bagasse, to biofuels such as ethanol has attracted significant interest as a potential element for transforming transport fuel supplies to totally renewable sources. However, the refractory nature of the cellulosic structure of lignocellulosic materials has impeded progress on developing an economic process, whereby the cellulose component may be effectively broken down to glucose monosaccharides and then purified to allow downstream fermentation. Ionic liquid (IL) treatment of lignocellulosic biomass has been shown to disrupt the crystalline structure of cellulose thus potentially enabling the cellulose to be more readily hydrolysed to monosaccharides. Furthermore, conventional hydrolysis of lignocellulosic materials yields byproducts that are inhibitors for efficient fermentation of the monosaccharides. However, selective extraction of monosaccharides from an aqueous/IL phase into an organic phase utilizing a combination of boronic acids and quaternary amines has shown promise as a purification process. Hydrolysis of sugar cane bagasse immersed in an aqueous solution with IL (1-ethyl-3-methylimidazolium acetate) was conducted at different pH and temperature below 100 ºC. It was found that the use of a high concentration of hydrochloric acid to acidify the solution inhibited the hydrolysis of bagasse. At high pH (i.e. basic conditions), using sodium hydroxide, catalyst yields were reduced for total reducing sugars (TRS) due to the rapid degradation of the sugars formed. For purification trials, a supported liquid membrane (SLM) apparatus was constructed, whereby a synthetic solution containing xylose and glucose in an aqueous IL phase was transported across a membrane impregnated with phenyl boronic acid/Aliquat 336 to an aqueous phase. The transport rate of xylose was generally higher than that of glucose indicating that a SLM scheme may not only be useful for purifying sugars from undesirable toxic compounds, but also for fractionating sugars to improve fermentation efficiency.

Keywords: biomass, bagasse, hydrolysis, monosaccharide, supported liquid membrane, purification

Procedia PDF Downloads 254
131 Heterogeneity of Genes Encoding the Structural Proteins of Avian Infectious Bronchitis Virus

Authors: Shahid Hussain Abro, Siamak Zohari, Lena H. M. Renström, Désirée S. Jansson, Faruk Otman, Karin Ullman, Claudia Baule

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Infectious bronchitis is an acute, highly contagious respiratory, nephropathogenic and reproductive disease of poultry that is caused by infectious bronchitis virus (IBV). The present study used a large data set of structural gene sequences, including newly generated ones and sequences available in the GenBank database to further analyze the diversity and to identify selective pressures and recombination spots. There were some deletions or insertions in the analyzed regions in isolates of the Italy-02 and D274 genotypes. Whereas, there were no insertions or deletions observed in the isolates of the Massachusetts and 4/91 genotype. The hypervariable nucleotide sequence regions spanned positions 152–239, 554–582, 686–737 and 802–912 in the S1 sub-unit of the all analyzed genotypes. The nucleotide sequence data of the E gene showed that this gene was comparatively unstable and subjected to a high frequency of mutations. The M gene showed substitutions consistently distributed except for a region between nucleotide positions 250–680 that remained conserved. The lowest variation in the nucleotide sequences of ORF5a was observed in the isolates of the D274 genotype. While, ORF5b and N gene sequences showed highly conserved regions and were less subjected to variation. Genes ORF3a, ORF3b, M, ORF5a, ORF5b and N presented negative selective pressure among the analyzed isolates. However, some regions of the ORFs showed favorable selective pressure(s). The S1 and E proteins were subjected to a high rate of mutational substitutions and non-synonymous amino acids. Strong signals of recombination breakpoints and ending break point were observed in the S and N genes. Overall, the results of this study revealed that very likely the strong selective pressures in E, M and the high frequency of substitutions in the S gene can probably be considered the main determinants in the evolution of IBV.

Keywords: IBV, avian infectious bronchitis, structural genes, genotypes, genetic diversity

Procedia PDF Downloads 435
130 X-Ray Crystallographic Studies on BPSL2418 from Burkholderia pseudomallei

Authors: Mona Alharbi

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Melioidosis has emerged as a lethal disease. Unfortunately, the molecular mechanisms of virulence and pathogenicity of Burkholderia pseudomallei remain unknown. However, proteomics research has selected putative targets in B. pseudomallei that might play roles in the B. pseudomallei virulence. BPSL 2418 putative protein has been predicted as a free methionine sulfoxide reductase and interestingly there is a link between the level of the methionine sulfoxide in pathogen tissues and its virulence. Therefore in this work, we describe the cloning expression, purification, and crystallization of BPSL 2418 and the solution of its 3D structure using X-ray crystallography. Also, we aimed to identify the substrate binding and reduced forms of the enzyme to understand the role of BPSL 2418. The gene encoding BPSL2418 from B. pseudomallei was amplified by PCR and reclone in pETBlue-1 vector and transformed into E. coli Tuner DE3 pLacI. BPSL2418 was overexpressed using E. coli Tuner DE3 pLacI and induced by 300μM IPTG for 4h at 37°C. Then BPS2418 purified to better than 95% purity. The pure BPSL2418 was crystallized with PEG 4000 and PEG 6000 as precipitants in several conditions. Diffraction data were collected to 1.2Å resolution. The crystals belonged to space group P2 21 21 with unit-cell parameters a = 42.24Å, b = 53.48Å, c = 60.54Å, α=γ=β= 90Å. The BPSL2418 binding MES was solved by molecular replacement with the known structure 3ksf using PHASER program. The structure is composed of six antiparallel β-strands and four α-helices and two loops. BPSL2418 shows high homology with the GAF domain fRMsrs enzymes which suggest that BPSL2418 might act as methionine sulfoxide reductase. The amino acids alignment between the fRmsrs including BPSL 2418 shows that the three cysteines that thought to catalyze the reduction are fully conserved. BPSL 2418 contains the three conserved cysteines (Cys⁷⁵, Cys⁸⁵ and Cys¹⁰⁹). The active site contains the six antiparallel β-strands and two loops where the disulfide bond formed between Cys⁷⁵ and Cys¹⁰⁹. X-ray structure of free methionine sulfoxide binding and native forms of BPSL2418 were solved to increase the understanding of the BPSL2418 catalytic mechanism.

Keywords: X-Ray Crystallography, BPSL2418, Burkholderia pseudomallei, Melioidosis

Procedia PDF Downloads 248
129 A Dissipative Particle Dynamics Study of a Capsule in Microfluidic Intracellular Delivery System

Authors: Nishanthi N. S., Srikanth Vedantam

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Intracellular delivery of materials has always proved to be a challenge in research and therapeutic applications. Usually, vector-based methods, such as liposomes and polymeric materials, and physical methods, such as electroporation and sonoporation have been used for introducing nucleic acids or proteins. Reliance on exogenous materials, toxicity, off-target effects was the short-comings of these methods. Microinjection was an alternative process which addressed the above drawbacks. However, its low throughput had hindered its adoption widely. Mechanical deformation of cells by squeezing them through constriction channel can cause the temporary development of pores that would facilitate non-targeted diffusion of materials. Advantages of this method include high efficiency in intracellular delivery, a wide choice of materials, improved viability and high throughput. This cell squeezing process can be studied deeper by employing simple models and efficient computational procedures. In our current work, we present a finite sized dissipative particle dynamics (FDPD) model to simulate the dynamics of the cell flowing through a constricted channel. The cell is modeled as a capsule with FDPD particles connected through a spring network to represent the membrane. The total energy of the capsule is associated with linear and radial springs in addition to constraint of the fixed area. By performing detailed simulations, we studied the strain on the membrane of the capsule for channels with varying constriction heights. The strain on the capsule membrane was found to be similar though the constriction heights vary. When strain on the membrane was correlated to the development of pores, we found higher porosity in capsule flowing in wider channel. This is due to localization of strain to a smaller region in the narrow constriction channel. But the residence time of the capsule increased as the channel constriction narrowed indicating that strain for an increased time will cause less cell viability.

Keywords: capsule, cell squeezing, dissipative particle dynamics, intracellular delivery, microfluidics, numerical simulations

Procedia PDF Downloads 140
128 Effect of Biostimulants to Control the Phelipanche ramosa L. Pomel in Processing Tomato Crop

Authors: G. Disciglio, G. Gatta, F. Lops, A. Libutti, A. Tarantino, E. Tarantino

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The experimental trial was carried out in open field at Foggia district (Apulia Region, Southern Italy), during the spring-summer season 2014, in order to evaluate the effect of four biostimulant products (RadiconÒ, Viormon plusÒ, LysodinÒ and SiaptonÒ 10L), compared with a control (no biostimulant), on the infestation of processing tomato crop (cv Dres) by the chlorophyll-lacking root parasite Phelipanche ramosa. Biostimulants consist in different categories of products (microbial inoculants, humic and fulvic acids, hydrolyzed proteins and aminoacids, seaweed extracts) which play various roles in plant growing, including the improvement of crop resistance and quali-quantitative characteristics of yield. The experimental trial was arranged according to a complete randomized block design with five treatments, each of one replicated three times. The processing tomato seedlings were transplanted on 5 May 2014. Throughout the crop cycle, P. ramosa infestation was assessed according to the number of emerged shoots (branched plants) counted in each plot, at 66, 78 and 92 day after transplanting. The tomato fruits were harvested at full-stage of maturity on 8 August 2014. From each plot, the marketable yield was measured and the quali-quantitative yield parameters (mean weight, dry matter content, colour coordinate, colour index and soluble solids content of the fruits) were determined. The whole dataset was tested according to the basic assumptions for the analysis of variance (ANOVA) and the differences between the means were determined using Tukey’s tests at the 5% probability level. The results of the study showed that none of the applied biostimulants provided a whole control of Phelipanche, although some positive effects were obtained from their application. To this respect, the RadiconÒ appeared to be the most effective in reducing the infestation of this root-parasite in tomato crop. This treatment also gave the higher tomato yield.

Keywords: biostimulant, control methods, Phelipanche ramosa, tomato crop

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127 Common Caper (Capparis Spinosa L.) From Oblivion and Neglect to the Interface of Medicinal Plants

Authors: Ahmad Alsheikh Kaddour

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Herbal medicine has been a long-standing phenomenon in Arab countries since ancient times because of its breadth and moderate temperament. Therefore, it possesses a vast natural and economic wealth of medicinal and aromatic herbs. This prompted ancient Egyptians and Arabs to discover and exploit them. The economic importance of the plant is not only from medicinal uses; it is a plant of high economic value for its various uses, especially in food, cosmetic and aromatic industries. It is also an ornamental plant and soil stabilization. The main objective of this research is to study the chemical changes that occur in the plant during the growth period, as well as the production of plant buds, which were previously considered unwanted plants. The research was carried out in the period 2021-2022 in the valley of Al-Shaflah (common caper), located in Qumhana village, 7 km north of Hama Governorate, Syria. The results of the research showed a change in the percentage of chemical components in the plant parts. The ratio of protein content and the percentage of fatty substances in fruits and the ratio of oil in the seeds until the period of harvesting of these plant parts improved, but the percentage of essential oils decreased with the progress of the plant growth, while the Glycosides content where improved with the plant aging. The production of buds is small, with dimensions as 0.5×0.5 cm, which is preferred for commercial markets, harvested every 2-3 days in quantities ranging from 0.4 to 0.5 kg in one cut/shrubs with 3 years’ age as average for the years 2021-2022. The monthly production of a shrub is between 4-5 kg per month. The productive period is 4 months approximately. This means that the seasonal production of one plant is 16-20 kg and the production of 16-20 tons per year with a plant density of 1,000 shrubs per hectare, which is the optimum rate of cultivation in the unit of mass, given the price of a kg of these buds is equivalent to 1 US $; however, this means that the annual output value of the locally produced hectare ranges from 16,000 US $ to 20,000 US $ for farmers. The results showed that it is possible to transform the cultivation of this plant from traditional random to typical areas cultivation, with a plant density of 1,000-1,100 plants per hectare according to the type of soil to obtain production of medicinal and nutritious buds, as well as, the need to pay attention to this national wealth and invest in the optimal manner, which leads to the acquisition of hard currency through export to support the national income.

Keywords: common caper, medicinal plants, propagation, medical, economic importance

Procedia PDF Downloads 72
126 Healthy Beverages Made from Grape Juice: Antioxidant, Energetic, and Isotonic Components

Authors: Yasmina Bendaali, Cristian Vaquero, Carlos Escott, Carmen González, Antonio Morata

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Consumer tendencies to healthy eating habits and request for organic beverages led to the production of new drinks from fruit juices as a source of nutrients and bioactive compounds. Grape juice is a rich source of sugars, organic acids, and phenolic compounds, which define its beneficial effect on health and the attractive sensory profile for consumers' choices (color, taste, flavor). Thus, grape juice was used as a source of sugars, avoiding the addition of sweeteners by diluting it with mineral water to obtain the sugar concentration recommended for isotonic drinks (6% to 8%) to provide energy during physical activities. In addition, phenolic compounds of grape juice are associated with many human health benefits, mainly antioxidant activity, which helps to prevent different diseases associated with oxidative stress, including cancers and cardiovascular and neurodegenerative diseases. Furthermore, physical exercise has been shown to increase the production of free radicals and other reactive oxygen species. Thus, athletes need to improve their antioxidant defense systems to prevent oxidative damage. Different studies have demonstrated the positive effect of grape juice consumption during physical activities, which improves antioxidant activity and performance, protects against oxidative damage, and reduces inflammation. Thus, the use of grape juice to develop isotonic drinks can provide isotonic drinks with antioxidant and biological activities in addition to their principal role of rehydration and replacement of minerals and carbohydrates during physical exercises. Moreover, attractive sensory characteristics, mainly color, which is provided by anthocyanin content, have a great contribution to making the drinks more natural and help to dispense the use of synthetic dyes in addition to the health benefits which will be a novel product in the field of healthy beverages responding on the demand of consumers for new, innovative, and healthy products.

Keywords: grape juice, isotonic, antioxidants, anthocyanins, natural, sport

Procedia PDF Downloads 77
125 A Study on the Effect of Different Climate Conditions on Time of Balance of Bleeding and Evaporation in Plastic Shrinkage Cracking of Concrete Pavements

Authors: Hasan Ziari, Hassan Fazaeli, Seyed Javad Vaziri Kang Olyaei, Asma Sadat Dabiri

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The presence of cracks in concrete pavements is a place for the ingression of corrosive substances, acids, oils, and water into the pavement and reduces its long-term durability and level of service. One of the causes of early cracks in concrete pavements is the plastic shrinkage. This shrinkage occurs due to the formation of negative capillary pressures after the equilibrium of the bleeding and evaporation rates at the pavement surface. These cracks form if the tensile stresses caused by the restrained shrinkage exceed the tensile strength of the concrete. Different climate conditions change the rate of evaporation and thus change the balance time of the bleeding and evaporation, which changes the severity of cracking in concrete. The present study examined the relationship between the balance time of bleeding and evaporation and the area of cracking in the concrete slabs using the standard method ASTM C1579 in 27 different environmental conditions by using continuous video recording and digital image analyzing. The results showed that as the evaporation rate increased and the balance time decreased, the crack severity significantly increased so that by reducing the balance time from the maximum value to its minimum value, the cracking area increased more than four times. It was also observed that the cracking area- balance time curve could be interpreted in three sections. An examination of these three parts showed that the combination of climate conditions has a significant effect on increasing or decreasing these two variables. The criticality of a single factor cannot cause the critical conditions of plastic cracking. By combining two mild environmental factors with a severe climate factor (in terms of surface evaporation rate), a considerable reduction in balance time and a sharp increase in cracking severity can be prevented. The results of this study showed that balance time could be an essential factor in controlling and predicting plastic shrinkage cracking in concrete pavements. It is necessary to control this factor in the case of constructing concrete pavements in different climate conditions.

Keywords: bleeding and cracking severity, concrete pavements, climate conditions, plastic shrinkage

Procedia PDF Downloads 146