Search results for: phorbol esters
Commenced in January 2007
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Edition: International
Paper Count: 123

Search results for: phorbol esters

33 Anticancer Lantadene Derivatives: Synthesis, Cytotoxic and Docking Studies

Authors: A. Monika, Manu Sharma, Hong Boo Lee, Richa Dhingra, Neelima Dhingra

Abstract:

Nuclear factor-κappa B serve as a molecular lynchpin that links persistent infections and chronic inflammation to increased cancer risk. Inflammation has been recognized as a hallmark and cause of cancer. Natural products present a privileged source of inspiration for chemical probe and drug design. Herbal remedies were the first medicines used by humans due to the many pharmacologically active secondary metabolites produced by plants. Some of the metabolites like Lantadene (pentacyclic triterpenoids) from the weed Lantana camara has been known to inhibit cell division and showed anti-antitumor potential. The C-3 aromatic esters of lantadenes were synthesized, characterized and evaluated for cytotoxicity and inhibitory potential against Tumor necrosis factor alpha-induced activation of Nuclear factor-κappa B in lung cancer cell line A549. The 3-methoxybenzoyloxy substituted lead analogue inhibited kinase activity of the inhibitor of nuclear factor-kappa B kinase in a single-digit micromolar concentration. At the same time, the lead compound showed promising cytotoxicity against A549 lung cancer cells with IC50 ( half maximal inhibitory concentration) of 0.98l µM. Further, molecular docking of 3-methoxybenzoyloxy substituted analogue against Inhibitor of nuclear factor-kappa B kinase (Protein data bank ID: 3QA8) showed hydrogen bonding interaction involving oxygen atom of 3-methoxybenzoyloxy with the Arginine-31 and Glutamine-110. Encouraging results indicate the Lantadene’s potential to be developed as anticancer agents.

Keywords: anticancer, lantadenes, pentacyclic triterpenoids, weed

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32 Study of Irritant and Anti-inflammatory Activity of Snuhi/Zaqqum (Euphorbia nerifolia) with Special Reference to Holy Quran and Ayurveda

Authors: Mohammed Khalil Ur Rahman, Pradnya Chigle, Bushra Farhen

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Indian mythology believes that Vedas are eternal treatises. Vedas are categorized into four divisions viz., Rigveda, Yajurveda, Samveda, Atharveda. All these spiritual classics not only deal with rituals and customs but also consist of inclusion of many references related to health. Out of these four, Atharveda deals with maximum principles pertaining to health sciences. Therefore, it is said that the science and the art of Ayurveda has developed from Atharveda. Ayurveda deals with many medicinal plants either as a single therapeutic use or in combination. One such medicinal plant is Snuhi (Euphorbia neriifolia Linn.) which finds its extensive importance along with Haridra and Apamargakshar, in the preparation of Ksharsutra which in turn is used for the treatment of Fistula in Ano. It is interesting to note that this plant Snuhi is also referred in Holy Quran as the Tree of Zaqqum advocated as the food for the sinners as a part of torment. The reference in Surat Ad-Dukhan is as follows: - 44:43-46. “Verily, the tree of Zaqqum will be the food of the sinners, Like boiling oil, it will boil in the bellies, like the boiling of scalding water.” The above verse implies that plant Snuhi/Zaqqum due to irritant property acts as a drastic purgative but at the same time it also possesses anti inflammatory properties in order to relieve the irritation. These properties of Zaqqum has been unfolded in the modern research which states that, Diterpene polycyclic esters are responsible for its toxic and irritant nature whereas; triterpenes are responsible for its anti inflammatory property. Present work will be an effort to review the concept of Quran about latex of the Tree of Zaqqum in terms of its phytochemistry and its therapeutic use in Ksharsutra pertaining to irritant and anti inflammatory property.

Keywords: ayurveda, Quran, zaqqum, ksharsutra, latex piles, inflammation

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31 Quality Evaluation of Grape Seed Oils of the Ionian Islands Based on GC-MS and Other Spectroscopic Techniques

Authors: I. Oikonomou, I. Lappa, D. Daferera, C. Kanakis, L. Kiokakis, K. Skordilis, A. Avramouli, E. Kalli, C. Pappas, P. A. Tarantilis, E. Skotti

Abstract:

Grape seeds are waste products of wineries and often referred to as an important agricultural and industrial waste product with the potential to be used in pharmaceutical, food, and cosmetic applications. In this study, grape seed oil from traditional Ionian varieties was examined for the determination of the quality and the characteristics of each variety. Initially, the fatty acid methyl ester (FAME) profiles were analyzed using Gas Chromatography-Mass Spectrometry, after transesterification. Furthermore, other quality parameters of the grape seed oils were determined by Spectroscopy techniques, UV-Vis and Raman included. Moreover, the antioxidant capacity of the oil was measured by 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assays and their antioxidant capacity expressed in Trolox equivalents. K and ΔΚ indices were measured in 232, 268, 270 nm, as an oil quality index. The results indicate that the air-dried grape seed total oil content ranged from 5.26 to 8.77% w/w, which is in accordance with the other grape seed varieties tested in similar studies. The composition of grape seed oil is predominated with linoleic and oleic fatty acids, with the linoleic fatty acid ranging from 53.68 to 69.95% and both the linoleic and oleic fatty acids totaling 78-82% of FAMEs, which is analogous to the fatty acid composition of safflower oil. The antioxidant assays ABTS and DPPH scored high, exhibiting that the oils have potential in the cosmetic and culinary businesses. Above that, our results demonstrate that Ionian grape seed oils have prospects that can go further than cosmetic or culinary use, into the pharmaceuticals industry. Finally, the reclamation of grape seeds from wineries waste stream is in accordance with the bio-economy strategic framework and contributes to environmental protection.

Keywords: antioxidant capacity, fatty acid methyl esters, grape seed oil, GC-MS

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30 Response Surface Methodology to Supercritical Carbon Dioxide Extraction of Microalgal Lipids

Authors: Yen-Hui Chen, Terry Walker

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As the world experiences an energy crisis, investing in sustainable energy resources is a pressing mission for many countries. Microalgae-derived biodiesel has attracted intensive attention as an important biofuel, and microalgae Chlorella protothecoides lipid is recognized as a renewable source for microalgae-derived biodiesel production. Supercritical carbon dioxide (SC-CO₂) is a promising green solvent that may potentially substitute the use of organic solvents for lipid extraction; however, the efficiency of SC-CO₂ extraction may be affected by many variables, including temperature, pressure and extraction time individually or in combination. In this study, response surface methodology (RSM) was used to optimize the process parameters, including temperature, pressure and extraction time, on C. protothecoides lipid yield by SC-CO₂ extraction. A second order polynomial model provided a good fit (R-square value of 0.94) for the C. protothecoides lipid yield. The linear and quadratic terms of temperature, pressure and extraction time—as well as the interaction between temperature and pressure—showed significant effects on lipid yield during extraction. The optimal lipid yield from the model was predicted as the temperature of 59 °C, the pressure of 350.7 bar and the extraction time 2.8 hours. Under these conditions, the experimental lipid yield (25%) was close to the predicted value. The principal fatty acid methyl esters (FAME) of C. protothecoides lipid-derived biodiesel were oleic acid methyl ester (60.1%), linoleic acid methyl ester (18.6%) and palmitic acid methyl ester (11.4%), which made up more than 90% of the total FAMEs. In summary, this study indicated that RSM was useful to characterize the optimization the SC-CO₂ extraction process of C. protothecoides lipid yield, and the second-order polynomial model could be used for predicting and describing the lipid yield very well. In addition, C. protothecoides lipid, extracted by SC-CO₂, was suggested as a potential candidate for microalgae-derived biodiesel production.

Keywords: Chlorella protothecoides, microalgal lipids, response surface methodology, supercritical carbon dioxide extraction

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29 Motif Search-Aided Screening of the Pseudomonas syringae pv. Maculicola Genome for Genes Encoding Tertiary Alcohol Ester Hydrolases

Authors: M. L. Mangena, N. Mokoena, K. Rashamuse, M. G. Tlou

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Tertiary alcohol ester (TAE) hydrolases are a group of esterases (EC 3.1.1.-) that catalyze the kinetic resolution of TAEs and as a result, they are sought-after for the production of optically pure tertiary alcohols (TAs) which are useful as building blocks for number biologically active compounds. What sets these enzymes apart is, the presence of a GGG(A)X-motif in the active site which appears to be the main reason behind their activity towards the sterically demanding TAEs. The genome of Pseudomonas syringae pv. maculicola (Psm) comprises a multitude of genes that encode esterases. We therefore, hypothesize that some of these genes encode TAE hydrolases. In this study, Psm was screened for TAE hydrolase activity using the linalyl acetate (LA) plate assay and a positive reaction was observed. As a result, the genome of Psm was screened for esterases with a GGG(A)X-motif using the motif search tool and two potential TAE hydrolase genes (PsmEST1 and 2, 1100 and 1000bp, respectively) were identified, PsmEST1 was amplified by PCR and the gene sequenced for confirmation. Analysis of the sequence data with the SingnalP 4.1 server revealed that the protein comprises a signal peptide (22 amino acid residues) on the N-terminus. Primers specific for the gene encoding the mature protein (without the signal peptide) were designed such that they contain NdeI and XhoI restriction sites for directional cloning of the PCR products into pET28a. The gene was expressed in E. coli JM109 (DE3) and the clones screened for TAE hydrolase activity using the LA plate assay. A positive clone was selected, overexpressed and the protein purified using nickel affinity chromatography. The activity of the esterase towards LA was confirmed using thin layer chromatography.

Keywords: hydrolases, tertiary alcohol esters, tertiary alcohols, screening, Pseudomonas syringae pv., maculicola genome, esterase activity, linalyl acetate

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28 Culturable Microbial Diversity and Adaptation Strategy in the Jutulsessen and Ahlmannryggen of Western Dronning Maud Land, Antarctica

Authors: Shiv Mohan Singh, Gwyneth Matcher

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To understand the culturable microbial composition and diversity patterns, soil samples were collected from inland nunataks of Jutulsessen and Ahlmannryggen ranges in Dronning Maud Land, Antarctica. 16S rRNA, ITS and the D1/D2 domain sequencing techniques were used for characterization of microbial communities of these geographical areas. The total 37 species of bacteria such as Arthrobacter agilis, Acinetobacter baumannii, Arthrobacter flavus, Arthrobacter ginsengisoli, Arthrobacter oxydans, Arthrobacter oryzae, Arthrobacter polychromogenes, Arthrobacter sulfonivorans, Bacillus altitudinis, Bacillus cereus, Bacillus paramycoides, Brevundimonas vesicularis, Brachybacterium rhamnosum, Curtobacterium luteum, Dermacoccus nishinomiyaensis, Dietzia aerolata, Janibacter indicus, Knoellia subterranean, Kocuria palustris, Kytococcus aerolatus, Lysinibacillus sphaericus, Microbacterium phyllosphaerae, Micrococcus yunnanensis, Methylobacterium rhodesianum, Moraxella osloensis, Paracoccus acridae, Pontibacter amylolyticus, Pseudomonas hunanensis, Pseudarthrobacter siccitolerans, Pseudarthrobacter phenanthrenivorans, Rhodococcus aerolatus, Rhodococcus sovatensis, Sphingomonas daechungensis, Sphingomonas sanguinis, Stenotrophomonas pavanii, Staphylococcus gallinarum, Staphylococcus arlettae and 9 species of fungi such as Candida davisiana, Cosmospora arxii, Geomyces destructans, Lecanicillium muscarium, Memnoniella humicola, Paecilomyces lilacinus, Pseudogymnoascus verrucosus, Phaeophlebiopsis ignerii and Thyronectria caraganae were recorded. Fatty acid methyl esters (FAME) analyses of representative species of each genus have shown predominance branched and unsaturated fatty acids indicate its adaptation strategy in Antarctic cold environment. To the best of our knowledge, this is the first record of culturable bacterial communities from Jutulsessen and Ahlmannryggen ranges in Western Dronning Maud Land, Antarctica.

Keywords: antarctica, microbe, adaptation, polar

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27 Comparison of Fatty Acids Composition of Three Commercial Fish Species Farmed in the Adriatic Sea

Authors: Jelka Pleadin, Greta Krešić, Tina Lešić, Ana Vulić, Renata Barić, Tanja Bogdanović, Dražen Oraić, Ana Legac, Snježana Zrnčić

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Fish has been acknowledged as an integral component of a well-balanced diet, providing a healthy source of energy, high-quality proteins, vitamins, essential minerals and, especially, n-3 long-chain polyunsaturated fatty acids (n-3 LC PUFA), mainly eicosapentaenoic acid (20:5 n-3 EPA), and docosahexaenoicacid, (22:6 n-3 DHA), whose pleiotropic effects in terms of health promotion and disease prevention have been increasingly recognised. In this study, the fatty acids composition of three commercially important farmed fish species: sea bream (Sparus aurata), sea bass (Dicentrarchus labrax) and dentex (Dentex dentex) was investigated. In total, 60 fish samples were retrieved during 2015 (n = 30) and 2016 (n = 30) from different locations in the Adriatic Sea. Methyl esters of fatty acids were analysed using gas chromatography (GC) with flame ionization detection (FID). The results show that the most represented fatty acid in all three analysed species is oleic acid (C18:1n-9, OA), followed by linoleic acid (C18:2n-6, LA) and palmitic acid (C16:0, PA). Dentex was shown to have two to four times higher eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid content as compared to sea bream and sea bass. The recommended n-6/n-3 ratio was determined in all fish species but obtained results pointed to statistically significant differences (p < 0.05) in fatty acid composition among the analysed fish species and their potential as a dietary source of valuable fatty acids. Sea bass and sea bream had a significantly higher proportion of n-6 fatty acids, while dentex had a significantly higher proportion of n-3 (C18:4n-3, C20:4n-3, EPA, DHA) fatty acids. A higher hypocholesterolaemic and hypercholesterolaemic fatty acids (HH) ratio was determined for sea bass and sea bream, which comes as the consequence of a lower share of SFA determined in these two species in comparison to dentex. Since the analysed fish species vary in their fatty acids composition consumption of diverse fish species would be advisable. Based on the established lipid quality indicators, dentex, a fish species underutilised by the aquaculture, seems to be a highly recommendable and important source of fatty acids recommended to be included into the human diet.

Keywords: dentex, fatty acids, farmed fish, sea bass, sea bream

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26 Algae for Wastewater Treatment and CO₂ Sequestration along with Recovery of Bio-Oil and Value Added Products

Authors: P. Kiran Kumar, S. Vijaya Krishna, Kavita Verma1, V. Himabindu

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Concern about global warming and energy security has led to increased biomass utilization as an alternative feedstock to fossil fuels. Biomass is a promising feedstock since it is abundant and cheap and can be transformed into fuels and chemical products. Microalgae biofuels are likely to have a much lower impact on the environment. Microalgae cultivation using sewage with industrial flue gases is a promising concept for integrated biodiesel production, CO₂ sequestration, and nutrients recovery. Autotrophic, Mixotrophic, and Heterotrophic are the three modes of cultivation for microalgae biomass. Several mechanical and chemical processes are available for the extraction of lipids/oily components from microalgae biomass. In organic solvent extraction methods, a prior drying of biomass and recovery of the solvent is required, which are energy-intensive. Thus, the hydrothermal process overcomes the drawbacks of conventional solvent extraction methods. In the hydrothermal process, the biomass is converted into oily components by processing in a hot, pressurized water environment. In this process, in addition to the lipid fraction of microalgae, other value-added products such as proteins, carbohydrates, and nutrients can also be recovered. In the present study was (Scenedesmus quadricauda) was isolated and cultivated in autotrophic, heterotrophic, and mixotrophically using sewage wastewater and industrial flue gas in batch and continuous mode. The harvested algae biomass from S. quadricauda was used for the recovery of lipids and bio-oil. The lipids were extracted from the algal biomass using sonication as a cell disruption method followed by solvent (Hexane) extraction, and the lipid yield obtained was 8.3 wt% with Palmitic acid, Oleic acid, and Octadeonoic acid as fatty acids. The hydrothermal process was also carried out for extraction of bio-oil, and the yield obtained was 18wt%. The bio-oil compounds such as nitrogenous compounds, organic acids, and esters, phenolics, hydrocarbons, and alkanes were obtained by the hydrothermal process of algal biomass. Nutrients such as NO₃⁻ (68%) and PO₄⁻ (15%) were also recovered along with bio-oil in the hydrothermal process.

Keywords: flue gas, hydrothermal process, microalgae, sewage wastewater, sonication

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25 Fatty Acids in Female's Gonads of the Red Sea Fish Rhabdosargus Sarba During the Spawning Season

Authors: Suhaila Qari, Samia Moharram, Safaa Alowaidi

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Objectives: To determine the fatty acids profiles in female fish, R. sarba from the Red Sea during the spawning season. Methods: Monthly individual Rhabdosargus sarba were obtained from Bangalah market in Jeddah, Red Sea and transported to the laboratory in ice aquarium. The total length, standard length and weight were measured, fishes were dissected. Ovaries were removed, weighed and 10 ml of concentrated hydrochloric acid were added to 10g of the ovary in a conical flask and immersed in boiling water until the sample was dissolved and the fat was seen to collect on the surface. The conical was cooled and the fat was extracted by shaking with 30 ml of diethyl ether. The extract was bowled after allowing the layers to separate into a weighed flask. The extraction was repeated three times more and distilled off the solvent then the fat dried at 100oC, cooled and weighed. Then 50 mg of lipid was put in a tube, 5 ml of methanolic sulphuric acid was added and 2 ml of benzene, the tube well closed and placed in water bath at 90oC for an hour and half. After cooling, 8 ml water and 5 ml petroleum was added shacked strongly and the ethereal layer was separated in a dry tube, evaporated to dryness. The fatty acid methyl esters were analyzed using a Hewlett Packard (HP 6890) chromatography, asplit /splitless injector and flame ionization detector (FID). Results: In female Rhabdosargus sarba, a total of 29 fatty acids detected in ovaries throughout the spawning season. The main fatty acid group in total lipid was saturated fatty acid (SFA, 28.9%), followed by 23.5% of polyunsaturated fatty acids (PUFA) and 12.9% of monounsaturated fatty acids (MUFA). The dominant SFA were palmitic and stearic, the major MUFA were palmitoleic and oleic, and the major PUFA were C18:2 and C22:2. During spawning stages no significant differences in total SFA, MUFA and PUFA, the highest value of SFA was in late spawning (36.78%). However, the highest value of MUFA and PUFA was in spawning (16.70% and 24.96% respectively). During spawning season there were a significant differences in total SFA between March (late spawning stage) and December (nearly ripe stage), (P < 0.05).

Keywords: sparidae, Rhabdosargus sarba, fish, fatty acids, spawning, gonads, red sea

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24 An Association Model to Correlate the Experimentally Determined Mixture Solubilities of Methyl 10-Undecenoate with Methyl Ricinoleate in Supercritical Carbon Dioxide

Authors: V. Mani Rathnam, Giridhar Madras

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Fossil fuels are depleting rapidly as the demand for energy, and its allied chemicals are continuously increasing in the modern world. Therefore, sustainable renewable energy sources based on non-edible oils are being explored as a viable option as they do not compete with the food commodities. Oils such as castor oil are rich in fatty acids and thus can be used for the synthesis of biodiesel, bio-lubricants, and many other fine industrial chemicals. There are several processes available for the synthesis of different chemicals obtained from the castor oil. One such process is the transesterification of castor oil, which results in a mixture of fatty acid methyl esters. The main products in the above reaction are methyl ricinoleate and methyl 10-undecenoate. To separate these compounds, supercritical carbon dioxide (SCCO₂) was used as a green solvent. SCCO₂ was chosen as a solvent due to its easy availability, non-toxic, non-flammable, and low cost. In order to design any separation process, the preliminary requirement is the solubility or phase equilibrium data. Therefore, the solubility of a mixture of methyl ricinoleate with methyl 10-undecenoate in SCCO₂ was determined in the present study. The temperature and pressure range selected for the investigation were T = 313 K to 333 K and P = 10 MPa to 18 MPa. It was observed that the solubility (mol·mol⁻¹) of methyl 10-undecenoate varied from 2.44 x 10⁻³ to 8.42 x 10⁻³ whereas it varied from 0.203 x 10⁻³ to 6.28 x 10⁻³ for methyl ricinoleate within the chosen operating conditions. These solubilities followed a retrograde behavior (characterized by the decrease in the solubility values with the increase in temperature) throughout the range of investigated operating conditions. An association theory model, coupled with regular solution theory for activity coefficients, was developed in the present study. The deviation from the experimental data using this model can be quantified using the average absolute relative deviation (AARD). The AARD% for the present compounds is 4.69 and 8.08 for methyl 10-undecenoate and methyl ricinoleate, respectively in a mixture of methyl ricinoleate and methyl 10-undecenoate. The maximum solubility enhancement of 32% was observed for the methyl ricinoleate in a mixture of methyl ricinoleate and methyl 10-undecenoate. The highest selectivity of SCCO₂ was observed to be 12 for methyl 10-undecenoate in a mixture of methyl ricinoleate and methyl 10-undecenoate.

Keywords: association theory, liquid mixtures, solubilities, supercritical carbon dioxide

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23 Identification of Bioactive Substances of Opuntia ficus-indica By-Products

Authors: N. Chougui, R. Larbat

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The first economic importance of Opuntia ficus-indica relies on the production of edible fruits. This food transformation generates a large amount of by-products (seeds and peels) in addition to cladodes produced by the plant. Several studies showed the richness of these products with bioactive substances like phenolics that have potential applications. Indeed, phenolics have been associated with protection against oxidation and several biological activities responsible of different pathologies. Consequently, there has been a growing interest in identifying natural antioxidants from plants. This study falls within the framework of the industrial exploitation of by-products of the plant. The study aims to investigate the metabolic profile of three by-products (cladodes, peel seeds) regarding total phenolic content by liquid chromatography coupled to mass spectrometry approach (LC-MSn). The byproducts were first washed, crushed and stored at negative temperature. The total phenolic compounds were then extracted by aqueous-ethanolic solvent in order to be quantified and characterized by LC-MS. According to the results obtained, the peel extract was the richest in phenolic compounds (1512.58 mg GAE/100 g DM) followed by the cladode extract (629.23 GAE/100 g DM) and finally by the seed extract (88.82 GAE/100 g DM) which is mainly used for its oil. The LC-MS analysis revealed diversity in phenolics in the three extracts and allowed the identification of hydroxybenzoic acids, hydroxycinnamic acids and flavonoids. The highest complexity was observed in the seed phenolic composition; more than twenty compounds were detected that belong to acids esters among which three feruloyl sucrose isomers. Sixteen compounds belonging to hydroxybenzoic acids, hydroxycinnamic acids and flavonoids were identified in the peel extract, whereas, only nine compounds were found in the cladode extract. It is interesting to highlight that the phenolic composition of the cladode extract was closer to that of the peel exact. However, from a quantitative viewpoint, the peel extract presented the highest amounts. Piscidic and eucomic acids were the two most concentrated molecules, corresponding to 271.3 and 121.6 mg GAE/ 100g DM respectively. The identified compounds were known to have high antioxidant and antiradical potential with the ability to inhibit lipid peroxidation and to exhibit a wide range of biological and therapeutic properties. The findings highlight the importance of using the Opuntia ficus-indica by-products.

Keywords: characterization, LC-MSn analysis, Opuntia ficus-indica, phenolics

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22 Desirable Fatty Acids in Meat of Cattle Fed Different Levels of Lipid-Based Diets

Authors: Tiago N. P. Valente, Erico S. Lima, Roberto O. Roça

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Introduction: Research has stimulated animal production studies on solutions to decrease the level of saturated fatty acids and increase unsaturated in foods of animal origin. The objective of this study was to determine the effect of the dietary inclusion of lipid-based diets on the fatty acid profiles from finishing cattle. Materials and Methods: The study was carried out in the Chapéu de Couro Farm in Aguaí/SP, Brazil. A group of 39 uncastrated Nellore cattle. Mean age of the animals was 36 months, and initial mean live weight was 494.1 ± 10.1. Animals were randomly assigned to one of three treatments, based on dry matter: feed with control diet 2.50% cottonseed, feed with 11.50% cottonseed, and feed with 3.13% cottonseed added of 1.77% protected lipid. Forage:concentrate ratio was 50:50 on a dry matter basis. Sugar cane chopped was used as forage. After 63 days mean final live weight was 577.01 kg ± 11.34. After slaughter, carcasses were identified and divided into two halves that were kept in a cold chamber for 24 hours at 2°C. Then, part of the M. longissimus thoracis of each animal was removed between the 12th and 13th rib of the left half carcass. The samples steaks were 2.5 cm thick and were identified and stored frozen in a freezer at -18°C. The analysis of methyl esters of fatty acids was carried out in a gas chromatograph. Desirable fatty acids (FADes) were determined by the sum of unsaturated fatty acids and stearic acid (C18:0). Results and Discussion: No differences (P>0.05) between the diets for the proportion of FADes in the meat of the animals in this study, according to the lipid sources used. The inclusion of protected fat or cottonseed in the diet did not change the proportion of FADes in the meat. The proportion mean of FADes in meat in the present study were: as pentadecanoic acid (C15:1 = 0.29%), palmitoleic acid (C16:1 = 4.26%), heptadecanoic acid (C17:1 = 0.07%), oleic acid (C18:1n9c = 37.32%), γ-linolenic acid (0.94%) and α-linolenic acid (1.04%), elaidic acid (C18:1n9t = 0.50%), eicosatrienoic acid (C20:3n3 = 0.03%), eicosapentaenoic acid (C20:5n3 = 0.04%), erucic acid (C22:1n9 = 0.89%), docosadienoic acid (C22:2 = 0.04%) and stearic acid (C18:0 = 21.53%). Conclusions: The add the cottonseed or protected lipid in diet is not affected fatty acids profiles the desirable fatty acids in meat. Acknowledgements: IFGoiano, FAPEG and CNPq (Brazil).

Keywords: beef quality, cottonseed, protected fat, unsaturated fatty acids

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21 Ultrastructural Characterization of Lipid Droplets of Rat Hepatocytes after Whole Body 60-Cobalt Gamma Radiation

Authors: Ivna Mororó, Lise P. Labéjof, Stephanie Ribeiro, Kely Almeida

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Lipid droplets (LDs) are normally presented in greater or lesser number in the cytoplasm of almost all eukaryotic and some prokaryotic cells. They are independent organelles composed of a lipid ester core and a surface phospholipid monolayer. As a lipid storage form, they provide an available source of energy for the cell. Recently it was demonstrated that they play an important role in other many cellular processes. Among the many unresolved questions about them, it is not even known how LDs is formed, how lipids are recruited to LDs and how they interact with the other organelles. Excess fat in the organism is pathological and often associated with the development of some genetic, hormonal or behavioral diseases. The formation and accumulation of lipid droplets in the cytoplasm can be increased by exogenous physical or chemical agents. It is well known that ionizing radiation affects lipid metabolism resulting in increased lipogenesis in cells, but the details of this process are unknown. To better understand the mode of formation of LDs in liver cells, we investigate their ultrastructural morphology after irradiation. For that, Wistar rats were exposed to whole body gamma radiation from 60-cobalt at various single doses. Samples of the livers were processed for analysis under a conventional transmission electron microscope. We found that when compared to controls, morphological changes in liver cells were evident at the higher doses of radiation used. It was detected a great number of lipid droplets of different sizes and homogeneous content and some of them merged each other. In some cells, it was observed diffused LDs, not limited by a monolayer of phospholipids. This finding suggests that the phospholipid monolayer of the LDs was disrupted by ionizing radiation exposure that promotes lipid peroxydation of endo membranes. Thus the absence of the phospholipid monolayer may prevent the realization of some cellular activities as follow: - lipid exocytosis which requires the merging of LDs membrane with the plasma membrane; - the interaction of LDs with other membrane-bound organelles such as the endoplasmic reticulum (ER), the golgi and mitochondria and; - lipolysis of lipid esters contained in the LDs which requires the presence of enzymes located in membrane-bound organelles as ER. All these impediments can contribute to lipid accumulation in the cytoplasm and the development of diseases such as liver steatosis, cirrhosis and cancer.

Keywords: radiobiology, hepatocytes, lipid metabolism, transmission electron microscopy

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20 Surface Display of Lipase on Yarrowia lipolytica Cells

Authors: Evgeniya Y. Yuzbasheva, Tigran V. Yuzbashev, Natalia I. Perkovskaya, Elizaveta B. Mostova

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Cell-surface display of lipase is of great interest as it has many applications in the field of biotechnology owing to its unique advantages: simplified product purification, and cost-effective downstream processing. One promising area of application for whole-cell biocatalysts with surface displayed lipase is biodiesel synthesis. Biodiesel is biodegradable, renewable, and nontoxic alternative fuel for diesel engines. Although the alkaline catalysis method has been widely used for biodiesel production, it has a number of limitations, such as rigorous feedstock specifications, complicated downstream processes, including removal of inorganic salts from the product, recovery of the salt-containing by-product glycerol, and treatment of alkaline wastewater. Enzymatic synthesis of biodiesel can overcome these drawbacks. In this study, Lip2p lipase was displayed on Yarrowia lipolytica cells via C- and N-terminal fusion variant. The active site of lipase is located near the C-terminus, therefore to prevent the activity loosing the insertion of glycine-serine linker between Lip2p and C-domains was performed. The hydrolytic activity of the displayed lipase reached 12,000–18,000 U/g of dry weight. However, leakage of enzyme from the cell wall was observed. In case of C-terminal fusion variant, the leakage was occurred due to the proteolytic cleavage within the linker peptide. In case of N-terminal fusion variant, the leaking enzyme was presented as three proteins, one of which corresponded to the whole hybrid protein. The calculated number of recombinant enzyme displayed on the cell surface is approximately 6–9 × 105 molecules per cell, which is close to the theoretical maximum (2 × 106 molecules/cell). Thus, we attribute the enzyme leakage to the limited space available on the cell surface. Nevertheless, cell-bound lipase exhibited greater stability to short-term and long-term temperature treatment than the native enzyme. It retained 74% of original activity at 60°C for 5 min of incubation, and 83% of original activity after incubation at 50°C during 5 h. Cell-bound lipase had also higher stability in organic solvents and detergents. The developed whole-cell biocatalyst was used for recycling biodiesel synthesis. Two repeated cycles of methanolysis yielded 84.1–% and 71.0–% methyl esters after 33–h and 45–h reactions, respectively.

Keywords: biodiesel, cell-surface display, lipase, whole-cell biocatalyst

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19 Hybrid Fermentation System for Improvement of Ergosterol Biosynthesis

Authors: Alexandra Tucaliuc, Alexandra C. Blaga, Anca I. Galaction, Lenuta Kloetzer, Dan Cascaval

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Ergosterol (ergosta-5,7,22-trien-3β-ol), also known as provitamin D2, is the precursor of vitamin D2 (ergocalciferol), because it is converted under UV radiation to this vitamin. The natural sources of ergosterol are mainly the yeasts (Saccharomyces sp., Candida sp.), but it can be also found in fungus (Claviceps sp.) or plants (orchids). In the yeasts cells, ergosterol is accumulated in membranes, especially in free form in the plasma membrane, but also as esters with fatty acids in membrane lipids. The chemical synthesis of ergosterol does not represent an efficient method for its production, in these circumstances, the most attractive alternative for producing ergosterol at larger-scale remains the aerobic fermentation using S. cerevisiae on glucose or by-products from agriculture of food industry as substrates, in batch or fed-batch operating systems. The aim of this work is to analyze comparatively the influence of aeration efficiency on ergosterol production by S. cerevisiae in batch and fed-batch fermentations, by considering different levels of mixing intensity, aeration rate, and n-dodecane concentration. The effects of the studied factors are quantitatively described by means of the mathematical correlations proposed for each of the two fermentation systems, valid both for the absence and presence of oxygen-vector inside the broth. The experiments were carried out in a laboratory stirred bioreactor, provided with computer-controlled and recorded parameters. n-Dodecane was used as oxygen-vector and the ergosterol content inside the yeasts cells has been considered at the fermentation moment related to the maximum concentration of ergosterol, 9 hrs for batch process and 20 hrs for fed-batch one. Ergosterol biosynthesis is strongly dependent on the dissolved oxygen concentration. The hydrocarbon concentration exhibits a significant influence on ergosterol production mainly by accelerating the oxygen transfer rate. Regardless of n-dodecane addition, by maintaining the glucose concentration at a constant level in the fed-batch process, the amount of ergosterol accumulated into the yeasts cells has been almost tripled. In the presence of hydrocarbon, the ergosterol concentration increased by over 50%. The value of oxygen-vector concentration corresponding to the maximum level of ergosterol depends mainly on biomass concentration, due to its negative influences on broth viscosity and interfacial phenomena of air bubbles blockage through the adsorption of hydrocarbon droplets–yeast cells associations. Therefore, for the batch process, the maximum ergosterol amount was reached for 5% vol. n-dodecane, while for the fed-batch process for 10% vol. hydrocarbon.

Keywords: bioreactors, ergosterol, fermentation, oxygen-vector

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18 Changes in Chromatographically Assessed Fatty Acid Profile during Technology of Dairy Products

Authors: Lina Lauciene, Vaida Andruleviciute, Ingrida Sinkeviciene, Mindaugas Malakauskas, Loreta Serniene

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Dairy product manufacturers constantly are looking for new markets for their production. And in most cases, the problem of product compliance with the composition requirements of foreign products is highlighted. This is especially true of the composition of milk fat in dairy products. It is well known that there are many factors such as feeding ratio, season, cow breed, stage of lactation that affect the fatty acid composition in milk. However, there is less evidence on the impact of the technological process on the composition of fatty acids in raw milk and products made from it. In this study the influence of the technological process on fat composition in 82% fat butter, 15% fat curd, 3.6% fat yogurt and 2.5% fat UHT milk was determined. The samples were collected at each stage of production, starting with raw milk and ending with the final product in the Lithuanian milk-processing company. Fatty acids methyl esters were quantified using a GC (Clarus 680, Perkin Elmer) equipped with flame ionization detector (FID) and a capillary column SP-2560, 100 m x 0.25 mm id x 0.20 µm. Fatty acids peaks were identified using Supelco® 37 Component FAME Mix. The concentration of each fatty acid was expressed in percent of the total fatty acid amount. In the case of UHT milk production, it was compared raw milk, cream, milk mixture, and UHT milk but significant differences were not estimated between these stages. Analyzing stages of the yogurt production (raw milk, pasteurized milk, and milk with a starter culture and yogurt), no significant changes were detected between stages as well. A slight difference was observed with C4:0 - a percentage of this fatty acid was less (p=0.053) in the final stage than in milk with the starter culture. During butter production, the composition of fatty acids in raw cream, buttermilk, and butter did not change significantly. Only C14:0 decreased in the butter then compared to buttermilk. The curd fatty acid analysis showed the increase of C6:0, C8:0, C10:0, C11:0, C12:0 C14:0 and C17:0 at the final stage when compared to raw milk, cream, milk mixture, and whey. Meantime the increase of C18:1n9c (in comparison with milk mixture and curd) and C18:2n6c (in comparison with raw milk, milk mixture, and curd) was estimated in cream. The results of this study suggest that the technological process did not affect the composition of fatty acids in UHT milk, yogurt, butter, and curd but had the impact on the concentration of individual fatty acids. In general, all of the fatty acids from the raw milk were converted into the final product, only some of them slightly changed the concentration. Therefore, in order to ensure an appropriate composition of certain fatty acids in the final product, producers must carefully choose the raw milk. Acknowledgment: This research was funded by Lithuanian Ministry of Agriculture (No. MT-17-13).

Keywords: dairy products, fat composition, fatty acids, technological process

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17 Single Cell Oil of Oleaginous Fungi from Lebanese Habitats as a Potential Feed Stock for Biodiesel

Authors: M. El-haj, Z. Olama, H. Holail

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Single cell oils (SCOs) accumulated by oleaginous fungi have emerged as a potential alternative feedstock for biodiesel production. Five fungal strains were isolated from the Lebanese environment namely Fusarium oxysporum, Mucor hiemalis, Penicillium citrinum, Aspergillus tamari, and Aspergillus niger that have been selected among 39 oleaginous strains for their potential ability to accumulate lipids (lipid content was more than 40% on dry weight basis). Wide variations were recorded in the environmental factors that lead to maximum lipid production by fungi under test and were cultivated under submerged fermentation on medium containing glucose as a carbon source. The maximum lipid production was attained within 6-8 days, at pH range 6-7, 24 to 48 hours age of seed culture, 4 to 6.107 spores/ml inoculum level and 100 ml culture volume. Eleven culture conditions were examined for their significance on lipid production using Plackett-Burman factorial design. Reducing sugars and nitrogen source were the most significant factors affecting lipid production process. Maximum lipid yield was noticed with 15.62, 14.48, 12.75, 13.68 and 20.41g/l for Fusarium oxysporum, Mucor hiemalis, Penicillium citrinum, Aspergillus tamari, and Aspergillus niger respectively. A verification experiment was carried out to examine model validation and revealed more than 94% validity. The profile of extracted lipids from each fungal isolate was studied using thin layer chromatography (TLC) indicating the presence of monoacylglycerols, diaacylglycerols, free fatty acids, triacylglycerols and sterol esters. The fatty acids profiles were also determined by gas-chromatography coupled with flame ionization detector (GC-FID). Data revealed the presence of significant amount of oleic acid (29-36%), palmitic acid (18-24%), linoleic acid (26.8-35%), and low amount of other fatty acids in the extracted fungal oils which indicate that the fatty acid profiles were quite similar to that of conventional vegetable oil. The cost of lipid production could be further reduced with acid-pretreated lignocellulotic corncob waste, whey and date molasses to be utilized as the raw material for the oleaginous fungi. The results showed that the microbial lipid from the studied fungi was a potential alternative resource for biodiesel production.

Keywords: agro-industrial waste products, biodiesel, fatty acid, single cell oil, Lebanese environment, oleaginous fungi

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16 A Study on the Shear-Induced Crystallization of Aliphatic-Aromatic Copolyester

Authors: Ramin Hosseinnezhad, Iurii Vozniak, Andrzej Galeski

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Shear-induced crystallization, originated from orientation of chains along the flow direction, is an inevitable part of most polymer processing technologies. It plays a dominant role in determining the final product properties and is affected by many factors such as shear rate, cooling rate, total strain, etc. Investigation of the shear-induced crystallization process become of great importance for preparation of nanocomposite, which requires crystallization of nanofibrous sheared inclusions at higher temperatures. Thus, the effects of shear time, shear rate, and also thermal condition of cooling on crystallization of two aliphatic-aromatic copolyesters have been investigated. This was performed using Linkam optical shearing system (CSS450) for both Ecoflex® F Blend C1200 produced by BASF and synthesized copolyester of butylene terephthalate and a mixture of butylene esters: adipate, succinate, and glutarate, (PBASGT), containing 60% of aromatic comonomer. Crystallization kinetics of these biodegradable copolyesters was studied at two different conditions of shearing. First, sample with a thickness of 60µm was heated to 60˚C above its melting point and subsequently subjected to different shear rates (100–800 sec-1) while cooling with specific rates. Second, the same type of sample was cooled down when shearing at constant temperature was finished. The intensity of transmitted depolarized light, recorded by a camera attached to the optical microscope, was used as a measure to follow the crystallization. Temperature dependencies of conversion degree of samples during cooling were collected and used to determine the half-temperature (Th), at which 50% conversion degree was reached. Shearing ecoflex films for 45 seconds with a shear rate of 100 sec-1 resulted in significant increase of Th from 56˚C to 70˚C. Moreover, the temperature range for the transition of molten samples to crystallized state decreased from 42˚C to 20˚C. Comparatively low shift of 10˚C in Th towards higher temperature was observed for PBASGT films at shear rate of 600 sec-1 for 45 seconds. However, insufficient melt flow strength and non-laminar flow due to Taylor vortices was a hindrance to reach more elevated Th at very high shear rates (600–800 sec-1). The shift in Th was smaller for the samples sheared at a constant temperature and subsequently cooled down. This may be attributed to the longer time gap between cessation of shearing and the onset of crystallization. The longer this time gap, the more possibility for crystal nucleus to re-melt at temperatures above Tm and for polymer chains to recoil and relax. It is found that the crystallization temperature, crystallization induction time and spherulite growth of aliphatic-aromatic copolyesters are dramatically influenced by both the cooling rate and the shear imposed during the process.

Keywords: induced crystallization, shear rate, aliphatic-aromatic copolyester, ecoflex

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15 Measurement of Fatty Acid Changes in Post-Mortem Belowground Carcass (Sus-scrofa) Decomposition: A Semi-Quantitative Methodology for Determining the Post-Mortem Interval

Authors: Nada R. Abuknesha, John P. Morgan, Andrew J. Searle

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Information regarding post-mortem interval (PMI) in criminal investigations is vital to establish a time frame when reconstructing events. PMI is defined as the time period that has elapsed between the occurrence of death and the discovery of the corpse. Adipocere, commonly referred to as ‘grave-wax’, is formed when post-mortem adipose tissue is converted into a solid material that is heavily comprised of fatty acids. Adipocere is of interest to forensic anthropologists, as its formation is able to slow down the decomposition process. Therefore, analysing the changes in the patterns of fatty acids during the early decomposition process may be able to estimate the period of burial, and hence the PMI. The current study concerned the investigation of the fatty acid composition and patterns in buried pig fat tissue. This was in an attempt to determine whether particular patterns of fatty acid composition can be shown to be associated with the duration of the burial, and hence may be used to estimate PMI. The use of adipose tissue from the abdominal region of domestic pigs (Sus-scrofa), was used to model the human decomposition process. 17 x 20cm piece of pork belly was buried in a shallow artificial grave, and weekly samples (n=3) from the buried pig fat tissue were collected over an 11-week period. Marker fatty acids: palmitic (C16:0), oleic (C18:1n-9) and linoleic (C18:2n-6) acid were extracted from the buried pig fat tissue and analysed as fatty acid methyl esters using the gas chromatography system. Levels of the marker fatty acids were quantified from their respective standards. The concentrations of C16:0 (69.2 mg/mL) and C18:1n-9 (44.3 mg/mL) from time zero exhibited significant fluctuations during the burial period. Levels rose (116 and 60.2 mg/mL, respectively) and fell starting from the second week to reach 19.3 and 18.3 mg/mL, respectively at week 6. Levels showed another increase at week 9 (66.3 and 44.1 mg/mL, respectively) followed by gradual decrease at week 10 (20.4 and 18.5 mg/mL, respectively). A sharp increase was observed in the final week (131.2 and 61.1 mg/mL, respectively). Conversely, the levels of C18:2n-6 remained more or less constant throughout the study. In addition to fluctuations in the concentrations, several new fatty acids appeared in the latter weeks. Other fatty acids which were detectable in the time zero sample, were lost in the latter weeks. There are several probable opportunities to utilise fatty acid analysis as a basic technique for approximating PMI: the quantification of marker fatty acids and the detection of selected fatty acids that either disappear or appear during the burial period. This pilot study indicates that this may be a potential semi-quantitative methodology for determining the PMI. Ideally, the analysis of particular fatty acid patterns in the early stages of decomposition could be an additional tool to the already available techniques or methods in improving the overall processes in estimating PMI of a corpse.

Keywords: adipocere, fatty acids, gas chromatography, post-mortem interval

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14 Evaluating the Effect of 'Terroir' on Volatile Composition of Red Wines

Authors: María Luisa Gonzalez-SanJose, Mihaela Mihnea, Vicente Gomez-Miguel

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The zoning methodology currently recommended by the OIVV as official methodology to carry out viticulture zoning studies and to define and delimit the ‘terroirs’ has been applied in this study. This methodology has been successfully applied on the most significant an important Spanish Oenological D.O. regions, such as Ribera de Duero, Rioja, Rueda and Toro, but also it have been applied around the world in Portugal, different countries of South America, and so on. This is a complex methodology that uses edaphoclimatic data but also other corresponding to vineyards and other soils’ uses The methodology is useful to determine Homogeneous Soil Units (HSU) to different scale depending on the interest of each study, and has been applied from viticulture regions to particular vineyards. It seems that this methodology is an appropriate method to delimit correctly the medium in order to enhance its uses and to obtain the best viticulture and oenological products. The present work is focused on the comparison of volatile composition of wines made from grapes grown in different HSU that coexist in a particular viticulture region of Castile-Lion cited near to Burgos. Three different HSU were selected for this study. They represented around of 50% of the global area of vineyards of the studied region. Five different vineyards on each HSU under study were chosen. To reduce variability factors, other criteria were also considered as grape variety, clone, rootstocks, vineyard’s age, training systems and cultural practices. This study was carried out during three consecutive years, then wine from three different vintage were made and analysed. Different red wines were made from grapes harvested in the different vineyards under study. Grapes were harvested to ‘Technological maturity’, which are correlated with adequate levels of sugar, acidity, phenolic content (nowadays named phenolic maturity), good sanitary stages and adequate levels of aroma precursors. Results of the volatile profile of the wines produced from grapes of each HSU showed significant differences among them pointing out a direct effect of the edaphoclimatic characteristic of each UHT on the composition of the grapes and then on the volatile composition of the wines. Variability induced by HSU co-existed with the well-known inter-annual variability correlated mainly with the specific climatic conditions of each vintage, however was most intense, so the wine of each HSU were perfectly differenced. A discriminant analysis allowed to define the volatiles with discriminant capacities which were 21 of the 74 volatiles analysed. Detected discriminant volatiles were chemical different, although .most of them were esters, followed by were superior alcohols and fatty acid of short chain. Only one lactone and two aldehydes were selected as discriminant variable, and no varietal aroma compounds were selected, which agree with the fact that all the wine were made from the same grape variety.

Keywords: viticulture zoning, terroir, wine, volatile profile

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13 Bio-Oil Compounds Sorption Enhanced Steam Reforming

Authors: Esther Acha, Jose Cambra, De Chen

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Hydrogen is considered an important energy vector for the 21st century. Nowadays there are some difficulties for hydrogen economy implantation, and one of them is the high purity required for hydrogen. This energy vector is still being mainly produced from fuels, from wich hydrogen is produced as a component of a mixture containing other gases, such as CO, CO2 and H2O. A forthcoming sustainable pathway for hydrogen is steam-reforming of bio-oils derived from biomass, e.g. via fast pyrolysis. Bio-oils are a mixture of acids, alcohols, aldehydes, esters, ketones, sugars phenols, guaiacols, syringols, furans, multi-functional compounds and also up to a 30 wt% of water. The sorption enhanced steam reforming (SESR) process is attracting a great deal of attention due to the fact that it combines both hydrogen production and CO2 separation. In the SESR process, carbon dioxide is captured by an in situ sorbent, which shifts the reversible reforming and water gas shift reactions to the product side, beyond their conventional thermodynamic limits, giving rise to a higher hydrogen production and lower cost. The hydrogen containing mixture has been obtained from the SESR of bio-oil type compounds. Different types of catalysts have been tested. All of them contain Ni at around a 30 wt %. Two samples have been prepared with the wet impregnation technique over conventional (gamma alumina) and non-conventional (olivine) supports. And a third catalysts has been prepared over a hydrotalcite-like material (HT). The employed sorbent is a commercial dolomite. The activity tests were performed in a bench-scale plant (PID Eng&Tech), using a stainless steel fixed bed reactor. The catalysts were reduced in situ in the reactor, before the activity tests. The effluent stream was cooled down, thus condensed liquid was collected and weighed, and the gas phase was analysed online by a microGC. The hydrogen yield, and process behavior was analysed without the sorbent (the traditional SR where a second purification step will be needed but that operates in steady state) and the SESR (where the purification step could be avoided but that operates in batch state). The influence of the support type and preparation method will be observed in the produced hydrogen yield. Additionally, the stability of the catalysts is critical, due to the fact that in SESR process sorption-desorption steps are required. The produced hydrogen yield and hydrogen purity has to be high and also stable, even after several sorption-desorption cycles. The prepared catalysts were characterized employing different techniques to determine the physicochemical properties of the fresh-reduced and used (after the activity tests) materials. The characterization results, together with the activity results show the influence of the catalysts preparation method, calcination temperature, or can even explain the observed yield and conversion.

Keywords: CO2 sorbent, enhanced steam reforming, hydrogen

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12 Carbon-Supported Pd Nano-Particles as Green Catalysts for the Production of Fuels from Biomass

Authors: Andrea Dragu, Solen Kinayyigit, Valerie Colliere, Karin Karin Philippot, Camelia Bala, Vasile I. Parvulescu

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The production of transportation fuels from biomass has gained a growing attention due to diminishing fossil fuel reserves, rising petroleum prices and increasing concern about global warming. In recent years, renewable hydrocarbons that are completely fungible with fossil fuels have been suggested to be efficiently produced by catalytic deoxygenation of fatty acids and their derivatives viadecarboxylation / decarbonylation. Several triglycerides (tall oil fatty acids) and saturated/unsaturated fatty acids and their corresponding esters were used as feedstocks. Their impact together with the influence of the reaction conditions and the catalyst composition on the nature of the reaction pathways of the deoxygenation of vegetable oils and their derivatives were recently reviewed. Following this state of the art the aim of the present study was the investigation of Pd NPs deposited onto mesoporous carbon supports as active and stable catalysts for the deoxygenation of oleic acid. The catalysts were prepared by the deposition of Pd NPs synthesised following an organometallic route on mesoporous carbons with different characteristics. Experiments were carried out under both batch and flow conditions. They demonstrated that under batch conditions (200 atm; 573K), the extent of the reaction depended, firstly, on the Pd loading and then on the metal dispersion and the oxidation state of palladium, both influenced by the way the support has been treated before the NPs deposition and by the preparation/stabilization methodology of Pd NPs. No aromatic compounds were detected in the reaction products but octadecanol and octadecane were observed in large extents. Under flow conditions (4 atm; 573 K), the conversion of stearic acid was superior to that observed in batch conditions. The product mixture contained over 20% heptadecane. No octadecanol, octadecane, and aromatic compounds were detected. The maxima in performances are obtained after only 0.5 h. After that, the yields in heptadecane suffer from a severe decrease until 3h reaction time. However, at that time, stopping feeding the reactor with oleic acid and flushing the catalyst only with mesitylene recovered the activity and the selectivity of the catalysts. With the complete removal of H2, the analysis revealed the presence of heptadecene in high excess compared to heptadecane (almost 7 to 1), thus suggesting decarbonylation as the main route. ICP-OES measurements indicated no leaching of palladium and simple washing of catalysts with mesitylene allowed recycling without any change in conversion or product distribution. Noteworthy, mesitylene as solvent exhibited no effect in this reaction. In conclusion, this study demonstrates the feasibility of such catalysts for the green production of fuels from biomass.

Keywords: fuels from biomass, green catalyst, Pd nano-particles , recycble catalyst

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11 Study of the Kinetics of Formation of Carboxylic Acids Using Ion Chromatography during Oxidation Induced by Rancimat of the Oleic Acid, Linoleic Acid, Linolenic Acid, and Biodiesel

Authors: Patrícia T. Souza, Marina Ansolin, Eduardo A. C. Batista, Antonio J. A. Meirelles, Matthieu Tubino

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Lipid oxidation is a major cause of the deterioration of the quality of the biodiesel, because the waste generated damages the engines. Among the main undesirable effects are the increase of viscosity and acidity, leading to the formation of insoluble gums and sediments which cause the blockage of fuel filters. The auto-oxidation is defined as the spontaneous reaction of atmospheric oxygen with lipids. Unsaturated fatty acids are usually the components affected by such reactions. They are present as free fatty acids, fatty esters and glycerides. To determine the oxidative stability of biodiesels, through the induction period, IP, the Rancimat method is used, which allows continuous monitoring of the induced oxidation process of the samples. During the oxidation of the lipids, volatile organic acids are produced as byproducts, in addition, other byproducts, including alcohols and carbonyl compounds, may be further oxidized to carboxylic acids. By the methodology developed in this work using ion chromatography, IC, analyzing the water contained in the conductimetric vessel, were quantified organic anions of carboxylic acids in samples subjected to oxidation induced by Rancimat. The optimized chromatographic conditions were: eluent water:acetone (80:20 v/v) with 0.5 mM sulfuric acid; flow rate 0.4 mL min-1; injection volume 20 µL; eluent suppressor 20 mM LiCl; analytical curve from 1 to 400 ppm. The samples studied were methyl biodiesel from soybean oil and unsaturated fatty acids standards: oleic, linoleic and linolenic. The induced oxidation kinetics curves were constructed by analyzing the water contained in the conductimetric vessels which were removed, each one, from the Rancimat apparatus at prefixed intervals of time. About 3 g of sample were used under the conditions of 110 °C and air flow rate of 10 L h-1. The water of each conductimetric Rancimat measuring vessel, where the volatile compounds were collected, was filtered through a 0.45 µm filter and analyzed by IC. Through the kinetic data of the formation of the organic anions of carboxylic acids, the formation rates of the same were calculated. The observed order of the rates of formation of the anions was: formate >>> acetate > hexanoate > valerate for the oleic acid; formate > hexanoate > acetate > valerate for the linoleic acid; formate >>> valerate > acetate > propionate > butyrate for the linolenic acid. It is possible to suppose that propionate and butyrate are obtained mainly from linolenic acid and that hexanoate is originated from oleic and linoleic acid. For the methyl biodiesel the order of formation of anions was: formate >>> acetate > valerate > hexanoate > propionate. According to the total rate of formation these anions produced during the induced degradation of the fatty acids can be assigned the order of reactivity: linolenic acid > linoleic acid >>> oleic acid.

Keywords: anions of carboxylic acids, biodiesel, ion chromatography, oxidation

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10 Interfacial Reactions between Aromatic Polyamide Fibers and Epoxy Matrix

Authors: Khodzhaberdi Allaberdiev

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In order to understand the interactions on the interface polyamide fibers and epoxy matrix in fiber- reinforced composites were investigated industrial aramid fibers: armos, svm, terlon using individual epoxy matrix components, epoxies: diglycidyl ether of bisphenol A (DGEBA), three- and diglycidyl derivatives of m, p-amino-, m, p-oxy-, o, m,p-carboxybenzoic acids, the models: curing agent, aniline and the compound, that depict of the structure the primary addition reaction the amine to the epoxy resin, N-di (oxyethylphenoxy) aniline. The chemical structure of the surface of untreated and treated polyamide fibers analyzed using Fourier transform infrared spectroscopy (FTIR). The impregnation of fibers with epoxy matrix components and N-di (oxyethylphenoxy) aniline has been carried out by heating 150˚C (6h). The optimum fiber loading is at 65%.The result a thermal treatment is the covalent bonds formation , derived from a combined of homopolymerization and crosslinking mechanisms in the interfacial region between the epoxy resin and the surface of fibers. The reactivity of epoxy resins on interface in microcomposites (MC) also depends from processing aids treated on surface of fiber and the absorbance moisture. The influences these factors as evidenced by the conversion of epoxy groups values in impregnated with DGEBA of the terlons: industrial, dried (in vacuum) and purified samples: 5.20 %, 4.65% and 14.10%, respectively. The same tendency for svm and armos fibers is observed. The changes in surface composition of these MC were monitored by X-ray photoelectron spectroscopy (XPS). In the case of the purified fibers, functional groups of fibers act as well as a catalyst and curing agent of epoxy resin. It is found that the value of the epoxy groups conversion for reinforced formulations depends on aromatic polyamides nature and decreases in the order: armos >svm> terlon. This difference is due of the structural characteristics of fibers. The interfacial interactions also examined between polyglycidyl esters substituted benzoic acids and polyamide fibers in the MC. It is found that on interfacial interactions these systems influences as well as the structure and the isomerism of epoxides. The IR-spectrum impregnated fibers with aniline showed that the polyamide fibers appreciably with aniline do not react. FTIR results of treated fibers with N-di (oxyethylphenoxy) aniline fibers revealed dramatically changes IR-characteristic of the OH groups of the amino alcohol. These observations indicated hydrogen bondings and covalent interactions between amino alcohol and functional groups of fibers. This result also confirms appearance of the exo peak on Differential Scanning Calorimetry (DSC) curve of the MC. Finally, the theoretical evaluation non-covalent interactions between individual epoxy matrix components and fibers has been performed using the benzanilide and its derivative contaning the benzimidazole moiety as a models of terlon and svm,armos, respectively. Quantum-topological analysis also demonstrated the existence hydrogen bond between amide group of models and epoxy matrix components.All the results indicated that on the interface polyamide fibers and epoxy matrix exist not only covalent, but and non-covalent the interactions during the preparation of MC.

Keywords: epoxies, interface, modeling, polyamide fibers

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9 Intraspecific Biochemical Diversity of Dalmatian Pyrethrum Across the Different Bioclimatic Regions of Its Natural Distribution Area

Authors: Martina Grdiša, Filip Varga, Nina Jeran, Ante Turudić, Zlatko Šatović

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Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevir.) Sch. Bip.) is a plant species that occurs naturally in the eastern Mediterranean. It is of immense economic importance as it synthesizes and accumulates the phytochemical compound pyrethrin. Pyrethrin consists of several monoterpene esters (pyrethrin I and II, cinerin I and II and jasmolin I and II), which have insecticidal and repellent activity through their synergistic action. In this study, 15 natural Dalmatian pyrethrum populations were sampled along their natural range in Croatia, Bosnia and Herzegovina and Montenegro to characterize and compare their pyrethrin profiles and to define the bioclimatic factors associated with the accumulation of each pyrethrin compound. Pyrethrins were extracted from the dried flower heads of Dalmatian pyrethrum using ultrasound-assisted extraction and the amount of each compound was quantified using high-performance liquid chromatography coupled to DAD-UV /VIS. The biochemical data were subjected to analysis of variance, correlation analysis and multivariate analysis. Quantitative variability within and among populations was found, with population P15 Vranjske Njive, Podgorica having the significantly highest pyrethrin I content (66.47% of total pyrethrin content), while the highest levels of total pyrethrin were found in P14 Budva (1.27% of dry flower weight; DW), followed by P08 Korčula (1.15% DW). Based on the environmental conditions at the sampling sites of the populations, five bioclimatic groups were distinguished, referred to as A, B, C, D, and E, each with rare chemical profile. The first group (A) consisted of the northern Adriatic population P01 Vrbnik, Krk and the population P06 Sevid - the coastal population of the central Adriatic, and generally differed significantly from the other bioclimatic groups by higher average jasmolin II values (2.13% of total pyrethrin). The second group (B) consisted of two central Adriatic island populations (P02 Telašćica, Dugi otok and P03 Žman, Dugi otok), while the remaining central Adriatic island populations were grouped in bioclimatic group C, which was characterized by the significantly highest average pyrethrin II (48.52% of total pyrethrin) and cinerin II (5.31% DW) content. The South Adriatic inland populations P10 Srđ and P11 Trebinje (Bosnia and Herzegovina), and the populations from Montenegro (P12 Grahovo, P13 Lovćen, P14 Budva and P15 Vranjske Njive, Podgorica) formed bioclimatic group E. This bioclimatic group was characterized by the highest average values for pyrethrin I (53.07 % of total pyrethrin), total pyrethrin content (1.06 % DW) and the ratio of pyrethrin I and II (1.85). Slightly lower values (although not significant) for the latter traits were detected in bioclimatic group D (southern Adriatic island populations P07 Vis, P08 Korčula and P09 Mljet). A weak but significant correlation was found between the levels of some pyrethrin compounds and bioclimatic variables (e.g., BIO03 Isothermality and BIO04 Temperature Seasonality), which explains part of the variability observed in the populations studied. This suggests the interconnection between bioclimatic variables and biochemical profiles either through the selection of adapted genotypes or through the ability of species to alter the expression of biochemical traits in response to environmental changes.

Keywords: biopesticides, biochemical variability, pyrethrin, Tanacetum cinerariifolium

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8 Design and Development of Graphene Oxide Modified by Chitosan Nanosheets Showing pH-Sensitive Surface as a Smart Drug Delivery System for Control Release of Doxorubicin

Authors: Parisa Shirzadeh

Abstract:

Drug delivery systems in which drugs are traditionally used, multi-stage and at specified intervals by patients, do not meet the needs of the world's up-to-date drug delivery. In today's world, we are dealing with a huge number of recombinant peptide and protean drugs and analogues of hormones in the body, most of which are made with genetic engineering techniques. Most of these drugs are used to treat critical diseases such as cancer. Due to the limitations of the traditional method, researchers sought to find ways to solve the problems of the traditional method to a large extent. Following these efforts, controlled drug release systems were introduced, which have many advantages. Using controlled release of the drug in the body, the concentration of the drug is kept at a certain level, and in a short time, it is done at a higher rate. Graphene is a natural material that is biodegradable, non-toxic, and natural compared to carbon nanotubes; its price is lower than carbon nanotubes and is cost-effective for industrialization. On the other hand, the presence of highly effective surfaces and wide surfaces of graphene plates makes it more effective to modify graphene than carbon nanotubes. Graphene oxide is often synthesized using concentrated oxidizers such as sulfuric acid, nitric acid, and potassium permanganate based on Hummer 1 method. In comparison with the initial graphene, the resulting graphene oxide is heavier and has carboxyl, hydroxyl, and epoxy groups. Therefore, graphene oxide is very hydrophilic and easily dissolves in water and creates a stable solution. On the other hand, because the hydroxyl, carboxyl, and epoxy groups created on the surface are highly reactive, they have the ability to work with other functional groups such as amines, esters, polymers, etc. Connect and bring new features to the surface of graphene. In fact, it can be concluded that the creation of hydroxyl groups, Carboxyl, and epoxy and in fact graphene oxidation is the first step and step in creating other functional groups on the surface of graphene. Chitosan is a natural polymer and does not cause toxicity in the body. Due to its chemical structure and having OH and NH groups, it is suitable for binding to graphene oxide and increasing its solubility in aqueous solutions. Graphene oxide (GO) has been modified by chitosan (CS) covalently, developed for control release of doxorubicin (DOX). In this study, GO is produced by the hummer method under acidic conditions. Then, it is chlorinated by oxalyl chloride to increase its reactivity against amine. After that, in the presence of chitosan, the amino reaction was performed to form amide transplantation, and the doxorubicin was connected to the carrier surface by π-π interaction in buffer phosphate. GO, GO-CS, and GO-CS-DOX characterized by FT-IR, RAMAN, TGA, and SEM. The ability to load and release is determined by UV-Visible spectroscopy. The loading result showed a high capacity of DOX absorption (99%) and pH dependence identified as a result of DOX release from GO-CS nanosheet at pH 5.3 and 7.4, which show a fast release rate in acidic conditions.

Keywords: graphene oxide, chitosan, nanosheet, controlled drug release, doxorubicin

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7 A Study on the Chemical Composition of Kolkheti's Sphagnum Peat Peloids to Evaluate the Perspective of Use in Medical Practice

Authors: Al. Tsertsvadze. L. Ebralidze, I. Matchutadze. D. Berashvili, A. Bakuridze

Abstract:

Peatlands are landscape elements, they are formed over a very long period by physical, chemical, biologic, and geologic processes. In the moderate zone of Caucasus, the Kolkheti lowlands are distinguished by the diversity of relictual plants, a high degree of endemism, orographic, climate, landscape, and other characteristics of high levels of biodiversity. The unique properties of the Kolkheti region lead to the formation of special, so-called, endemic peat peloids. The composition and properties of peloids strongly depend on peat-forming plants. Peat is considered a unique complex of raw materials, which can be used in different fields of the industry: agriculture, metallurgy, energy, biotechnology, chemical industry, health care. They are formed in permanent wetland areas. As a result of decay, higher plants remain in the anaerobic area, with the participation of microorganisms. Peat mass absorbs soil and groundwater. Peloids are predominantly rich with humic substances, which are characterized by high biological activity. Humic acids stimulate enzymatic activity, regenerative processes, and have anti-inflammatory activity. Objects of the research were Kolkheti peat peloids (Ispani, Anaklia, Churia, Chirukhi, Peranga) possessing different formation phases. Due to specific physical and chemical properties of research objects, the aim of the research was to develop analytical methods in order to study the chemical composition of the objects. The research was held using modern instrumental methods of analysis: Ultraviolet-visible spectroscopy and Infrared spectroscopy, Scanning Electron Microscopy, Centrifuge, dry oven, Ultraturax, pH meter, fluorescence spectrometer, Gas chromatography-mass spectrometry (GC-MS/MS), Gas chromatography. Based on the research ration between organic and inorganic substances, the spectrum of micro and macro elements, also the content of minerals was determined. The content of organic nitrogen was determined using the Kjeldahl method. The total composition of amino acids was studied by a spectrophotometric method using standard solutions of glutamic and aspartic acids. Fatty acid was determined using GC (Gas chromatography). Based on the obtained results, we can conclude that the method is valid to identify fatty acids in the research objects. The content of organic substances in the research objects was held using GC-MS. Using modern instrumental methods of analysis, the chemical composition of research objects was studied. Each research object is predominantly reached with a broad spectrum of organic (fatty acids, amino acids, carbocyclic and heterocyclic compounds, organic acids and their esters, steroids) and inorganic (micro and macro elements, minerals) substances. Modified methods used in the presented research may be utilized for the evaluation of cosmetological balneological and pharmaceutical means prepared on the base of Kolkheti's Sphagnum Peat Peloids.

Keywords: modern analytical methods, natural resources, peat, chemistry

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6 Key Aroma Compounds as Predictors of Pineapple Sensory Quality

Authors: Jenson George, Thoa Nguyen, Garth Sanewski, Craig Hardner, Heather Eunice Smyth

Abstract:

Pineapple (Ananas comosus), with its unique sweet flavour, is one of the most popular tropical, non-climacteric fruits consumed worldwide. It is also the third most important tropical fruit in world production. In Australia, 99% of the pineapple production is from the Queensland state due to the favourable subtropical climatic conditions. The flavourful fruit is known to contain around 500 volatile organic compounds (VOC) at varying concentrations and greatly contribute to the flavour quality of pineapple fruit by providing distinct aroma sensory properties that are sweet, fruity, tropical, pineapple-like, caramel-like, coconut-like, etc. The aroma of pineapple is one of the important factors attracting consumers and strengthening the marketplace. To better understand the aroma of Australian-grown pineapples, the matrix-matched Gas chromatography–mass spectrometry (GC-MS), Head Space - Solid-phase microextraction (HS-SPME), Stable-isotope dilution analysis (SIDA) method was developed and validated. The developed method represents a significant improvement over current methods with the incorporation of multiple external reference standards, multiple isotopes labeled internal standards, and a matching model system of pineapple fruit matrix. This method was employed to quantify 28 key aroma compounds in more than 200 genetically diverse pineapple varieties from a breeding program. The Australian pineapple cultivars varied in content and composition of free volatile compounds, which were predominantly comprised of esters, followed by terpenes, alcohols, aldehydes, and ketones. Using selected commercial cultivars grown in Australia, and by employing the sensorial analysis, the appearance (colour), aroma (intensity, sweet, vinegar/tang, tropical fruits, floral, coconut, green, metallic, vegetal, fresh, peppery, fermented, eggy/sulphurous) and texture (crunchiness, fibrousness, and juiciness) were obtained. Relationships between sensory descriptors and volatiles were explored by applying multivariate analysis (PCA) to the sensorial and chemical data. The key aroma compounds of pineapple exhibited a positive correlation with corresponding sensory properties. The sensory and volatile data were also used to explore genetic diversity in the breeding population. GWAS was employed to unravel the genetic control of the pineapple volatilome and its interplay with fruit sensory characteristics. This study enhances our understanding of pineapple aroma (flavour) compounds, their biosynthetic pathways and expands breeding option for pineapple cultivars. This research provides foundational knowledge to support breeding programs, post-harvest and target market studies, and efforts to optimise the flavour of commercial pineapple varieties and their parent lines to produce better tasting fruits for consumers.

Keywords: Ananas comosus, pineapple, flavour, volatile organic compounds, aroma, Gas chromatography–mass spectrometry (GC-MS), Head Space - Solid-phase microextraction (HS-SPME), Stable-isotope dilution analysis (SIDA).

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5 Applications of Polyvagal Theory for Trauma in Clinical Practice: Auricular Acupuncture and Herbology

Authors: Aurora Sheehy, Caitlin Prince

Abstract:

Within current orthodox medical protocols, trauma and mental health issues are deemed to reside within the realm of cognitive or psychological therapists and are marginalised in these areas, in part due to limited drugs option available, mostly manipulating neurotransmitters or sedating patients to reduce symptoms. By contrast, this research presents examples from the clinical practice of how trauma can be assessed and treated physiologically. Adverse Childhood Experiences (ACEs) are a tally of different types of abuse and neglect. It has been used as a measurable and reliable predictor of the likelihood of the development of autoimmune disease. It is a direct way to demonstrate reliably the health impact of traumatic life experiences. A second assessment tool is Allostatic Load, which refers to the cumulative effects that chronic stress has on mental and physical health. It records the decline of an individual’s physiological capacity to cope with their experience. It uses a specific grouping of serum testing and physical measures. It includes an assessment of neuroendocrine, cardiovascular, immune and metabolic systems. Allostatic load demonstrates the health impact that trauma has throughout the body. It forms part of an initial intake assessment in clinical practice and could also be used in research to evaluate treatment. Examining medicinal plants for their physiological, neurological and somatic effects through the lens of Polyvagal theory offers new opportunities for trauma treatments. In situations where Polyvagal theory recommends activities and exercises to enable parasympathetic activation, many herbs that affect Effector Memory T (TEM) cells also enact these responses. Traditional or Indigenous European herbs show the potential to support the polyvagal tone, through multiple mechanisms. As the ventral vagal nerve reaches almost every major organ, plants that have actions on these tissues can be understood via their polyvagal actions, such as monoterpenes as agents to improve respiratory vagal tone, cyanogenic glycosides to reset polyvagal tone, volatile oils rich in phenyl methyl esters improve both sympathetic and parasympathetic tone, bitters activate gut function and can strongly promote parasympathetic regulation. Auricular Acupuncture uses a system of somatotopic mapping of the auricular surface overlaid with an image of an inverted foetus with each body organ and system featured. Given that the concha of the auricle is the only place on the body where the Vagus Nerve neurons reach the surface of the skin, several investigators have evaluated non-invasive, transcutaneous electrical nerve stimulation (TENS) at auricular points. Drawn from an interdisciplinary evidence base and developed through clinical practice, these assessment and treatment tools are examples of practitioners in the field innovating out of necessity for the best outcomes for patients. This paper draws on case studies to direct future research.

Keywords: polyvagal, auricular acupuncture, trauma, herbs

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4 Reproductive Biology and Lipid Content of Albacore Tuna (Thunnus alalunga) in the Western Indian Ocean

Authors: Zahirah Dhurmeea, Iker Zudaire, Heidi Pethybridge, Emmanuel Chassot, Maria Cedras, Natacha Nikolic, Jerome Bourjea, Wendy West, Chandani Appadoo, Nathalie Bodin

Abstract:

Scientific advice on the status of fish stocks relies on indicators that are based on strong assumptions on biological parameters such as condition, maturity and fecundity. Currently, information on the biology of albacore tuna, Thunnus alalunga, in the Indian Ocean is scarce. Consequently, many parameters used in stock assessment models for Indian Ocean albacore originate largely from other studied stocks or species of tuna. Inclusion of incorrect biological data in stock assessment models would lead to inappropriate estimates of stock status used by fisheries manager’s to establish future catch allowances. The reproductive biology of albacore tuna in the western Indian Ocean was examined through analysis of the sex ratio, spawning season, length-at-maturity (L50), spawning frequency, fecundity and fish condition. In addition, the total lipid content (TL) and lipid class composition in the gonads, liver and muscle tissues of female albacore during the reproductive cycle was investigated. A total of 923 female and 867 male albacore were sampled from 2013 to 2015. A bias in sex-ratio was found in favour of females with fork length (LF) <100 cm. Using histological analyses and gonadosomatic index, spawning was found to occur between 10°S and 30°S, mainly to the east of Madagascar from October to January. Large females contributed more to reproduction through their longer spawning period compared to small individuals. The L50 (mean ± standard error) of female albacore was estimated at 85.3 ± 0.7 cm LF at the vitellogenic 3 oocyte stage maturity threshold. Albacore spawn on average every 2.2 days within the spawning region and spawning months from November to January. Batch fecundity varied between 0.26 and 2.09 million eggs and the relative batch fecundity (mean  standard deviation) was estimated at 53.4 ± 23.2 oocytes g-1 of somatic-gutted weight. Depending on the maturity stage, TL in ovaries ranged from 7.5 to 577.8 mg g-1 of wet weight (ww) with different proportions of phospholipids (PL), wax esters (WE), triacylglycerol (TAG) and sterol (ST). The highest TL were observed in immature (mostly TAG and PL) and spawning capable ovaries (mostly PL, WE and TAG). Liver TL varied from 21.1 to 294.8 mg g-1 (ww) and acted as an energy (mainly TAG and PL) storage prior to reproduction when the lowest TL was observed. Muscle TL varied from 2.0 to 71.7 g-1 (ww) in mature females without a clear pattern between maturity stages, although higher values of up to 117.3 g-1 (ww) was found in immature females. TL results suggest that albacore could be viewed predominantly as a capital breeder relying mostly on lipids stored before the onset of reproduction and with little additional energy derived from feeding. This study is the first one to provide new information on the reproductive development and classification of albacore in the western Indian Ocean. The reproductive parameters will reduce uncertainty in current stock assessment models which will eventually promote sustainability of the fishery.

Keywords: condition, size-at-maturity, spawning behaviour, temperate tuna, total lipid content

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