Search results for: amino acids
727 Genome-Wide Analysis of BES1/BZR1 Gene Family in Five Plant Species
Authors: Jafar Ahmadi, Zhohreh Asiaban, Sedigheh Fabriki Ourang
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Brassinosteroids (BRs) regulate cell elongation, vascular differentiation, senescence and stress responses. BRs signal through the BES1/BZR1 family of transcription factors, which regulate hundreds of target genes involved in this pathway. In this research a comprehensive genome-wide analysis was carried out in BES1/BZR1 gene family in Arabidopsis thaliana, Cucumis sativus, Vitis vinifera, Glycin max, and Brachypodium distachyon. Specifications of the desired sequences, dot plot and hydropathy plot were analyzed in the protein and genome sequences of five plant species. The maximum amino acid length was attributed to protein sequence Brdic3g with 374aa and the minimum amino acid length was attributed to protein sequence Gm7g with 163aa. The maximum Instability index was attributed to protein sequence AT1G19350 equal with 79.99 and the minimum Instability index was attributed to protein sequence Gm5g equal with 33.22. Aliphatic index of these protein sequences ranged from 47.82 to 78.79 in Arabidopsis thaliana, 49.91 to 57.50 in Vitis vinifera, 55.09 to 82.43 in Glycin max, 54.09 to 54.28 in Brachypodium distachyon 55.36 to 56.83 in Cucumis sativus. Overall, data obtained from our investigation contributes a better understanding of the complexity of the BES1/BZR1 gene family and provides the first step towards directing future experimental designs to perform systematic analysis of the functions of the BES1/BZR1 gene family.Keywords: BES1/BZR1, brassinosteroids, phylogenetic analysis, transcription factor
Procedia PDF Downloads 331726 Peach as a Potential Functional Food: Biological Activity and Important Phenolic Compound Source
Authors: Luís R. Silva, Catarina Bento, Ana C. Gonçalves, Fábio Jesus, Branca M. Silva
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Nowadays, the general population is more and more concerned about nutrition and the health implications of an unbalanced diet. Current knowledge regarding the health benefits and antioxidant properties of certain foods such as fruits and vegetables has gained the interest of both the general public and scientific community. Peach (Prunus persica (L.) Batsch) is one of the most consumed fruits worldwide, with low sugar contents and a broad range of nutrients essential to the normal functioning of the body. Six different peach cultivars from the Fundão region in Portugal were evaluated regarding their phenolic composition by LC-DAD and biological activity. The prepared extracts’ capacity to scavenge free-radicals was tested through the stable free radical DPPH• and nitric oxide (•NO). Additionally, antidiabetic potential and protective effects against peroxyl radical (ROO•) induced damage to erythrocytes were also tested. LC-DAD analysis allowed the identification of 17 phenolic compounds, among which 5-O-caffeoylquinic acids and 3-O-caffeoylquinic acids are pointed out as the most abundant. Regarding the antioxidant activity, all cultivars displayed concentration-dependent free-radical scavenging activity against both nitrogen species and DPPH•. In respect to α-glucosidase inhibitory activity, Royal Magister and Royal Glory presented the highest inhibitory activity (IC50 = 11.7 ± 1.4 and 17.1 ± 1.7 μg/mL, respectively), nevertheless all six cultivars presented higher activity than the control acarbose. As for the protective effect of Royal Lu extract on the oxidative damage induced in erythrocytes by ROO•, the results were quite promising showing inhibition IC50 values of 110.0 ± 4.5 μg/mL and 83.8 ± 6.5 μg/mL for hemolysis and hemoglobin oxidation, respectively. The demonstrated activity is of course associated to the peaches’ phenolic profile, rich in phenolic acids and flavonoids with high hydrogen donating capacity. These compounds have great industrial interest for the manufacturing of natural products. The following step would naturally be the extraction and isolation from the plant tissues and large-scale production through biotechnology techniques.Keywords: antioxidants, functional food, phenolic compounds, peach
Procedia PDF Downloads 284725 Using Human-Digestive Simulator to Harbor Encapsulated Lactobacillus casei 01 along with Pasteurized-Purple-Rice Drinks for Examination of the Health-Promoting Effects
Authors: Srivilai Worametrachanon, Arunee Apichartsrangkoon, Jiranat Techarang, Boonrak Phanchaisri
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A human-digestive simulator consisted of four colon compartments, i.e., stomach, small intestine, proximal colon and distal colon used to harbor L. casei 01 plus either pasteurized ordinary-purple-rice drinks or germinated-purple-rice drinks. Accordingly, three treatment compositions had been set up and the effects of treatments on colon bacterial communities including their by-products were thoroughly examined. L. casei 01 plus purple-rice drinks gave rise to significantly high formation (P ≤ 0.05) of short-chain-fatty acids (SCFA) of which highest acetic acid was found followed by propionic and butyric acids, while the germinated-rice drink showed the greatest impact. Moreover, the effect was more pronounced upon prolonged fermentation. In addition, the influence of treatments on colon microbes was also demonstrated. Accordingly, desirable bacteria including colon Lactobacilli and Bifidobacteria were significantly increased (P ≤ 0.05) in both colons in comparison with the control and the effect was more prominent after adding purple-rice drink. On the other hand, undesirable Clostridia and coliforms were apparently diminished by the influence of treatment conditions, in which both compartments exhibited similar results.Keywords: human-digestive simulator, Lactobacillus casei 01, Pasteurized-purple-rice drinks
Procedia PDF Downloads 217724 Liquid-Liquid Extraction of Uranium (VI) from Aqueous Solution Using 1-Hydroxyalkylidene-1,1-Diphosphonic Acids
Authors: Mustapha Bouhoun Ali, Ahmed Yacine Badjah Hadj Ahmed, Mouloud Attou, Abdel Hamid Elias, Mohamed Amine Didi
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The extraction of uranium(VI) from aqueous solutions has been investigated using 1-hydroxyhexadecylidene-1,1-diphosphonic acid (HHDPA) and 1-hydroxydodecylidene-1,1-diphosphonic acid (HDDPA), which were synthesized and characterized by elemental analysis and by FT-IR, 1H NMR, 31P NMR spectroscopy. In this paper, we propose a tentative assignment for the shifts of those two ligands and their specific complexes with uranium(VI). We carried out the extraction of uranium(VI) by HHDPA and HDDPA from [carbon tetrachloride + 2-octanol (v/v: 90%/10%)] solutions. Various factors such as contact time, pH, organic/aqueous phase ratio and extractant concentration were considered. The optimum conditions obtained were: contact time = 20 min, organic/aqueous phase ratio = 1, pH value = 3.0 and extractant concentration = 0.3M. The extraction yields are more significant in the case of the HHDPA which is equipped with a hydrocarbon chain, longer than that of the HDDPA. Logarithmic plots of the uranium(VI) distribution ratio vs. pHeq and the extractant concentration showed that the ratio of extractant to extracted uranium(VI) (ligand/metal) is 2:1. The formula of the complex of uranium(VI) with the HHDPA and the DHDPA is UO2(H3L)2 (HHDPA and DHDPA are denoted as H4L). A spectroscopic analysis has showed that coordination of uranium(VI) takes place via oxygen atoms.Keywords: liquid-liquid extraction, uranium(VI), 1-hydroxyalkylidene-1, 1-diphosphonic acids, HHDPA, HDDPA, aqueous solution
Procedia PDF Downloads 525723 Optimization of Bio-Diesel Production from Rubber Seed Oils
Authors: Pawit Tangviroon, Apichit Svang-Ariyaskul
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Rubber seed oil is an attractive alternative feedstock for biodiesel production because it is not related to food-chain plant. Rubber seed oil contains large amount of free fatty acids, which causes problem in biodiesel production. Free fatty acids can react with alkaline catalyst in biodiesel production. Acid esterification is used as pre-treatment to convert unwanted compound to desirable biodiesel. Phase separation of oil and methanol occurs at low ratio of methanol to oil and causes low reaction rate and conversion. Acid esterification requires large excess of methanol in order to increase the miscibility of methanol in oil and accordingly, it is a more expensive separation process. In this work, the kinetics of esterification of rubber seed oil with methanol is developed from available experimental results. Reactive distillation process was designed by using Aspen Plus program. The effects of operating parameters such as feed ratio, molar reflux ratio, feed temperature, and feed stage are investigated in order to find the optimum conditions. Results show that the reactive distillation process is proved to be better than conventional process. It consumes less feed methanol and less energy while yielding higher product purity than the conventional process. This work can be used as a guideline for further development to industrial scale of biodiesel production using reactive distillation.Keywords: biodiesel, reactive distillation, rubber seed oil, transesterification
Procedia PDF Downloads 339722 Molecular Characterisation and Expression of Glutathione S-Transferase of Fasciola Gigantica
Authors: J. Adeppa, S. Samanta, O. K. Raina
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Fasciolosis is a widespread economically important parasitic infection throughout the world caused by Fasciola hepatica and F. gigantica. In order to identify novel immunogen conferring significant protection against fasciolosis, currently, research has been focused on the defined antigens viz. glutathione S-transferase, fatty acid binding protein, cathepsin-L, fluke hemoglobin, paramyosin, myosin and F. hepatica- Kunitz Type Molecule. Among various antigens, GST which plays a crucial role in detoxification processes, i.e. phase II defense mechanism of this parasite, has a unique position as a novel vaccine candidate and a drug target in the control of this disease. For producing the antigens in large quantities and their purification to complete homogeneity, the recombinant DNA technology has become an important tool to achieve this milestone. RT- PCR was carried out using F. gigantica total RNA as template, and an amplicon of 657 bp GST gene was obtained. TA cloning vector was used for cloning of this gene, and the presence of insert was confirmed by blue-white selection for recombinant colonies. Sequence analysis of the present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin (accession no. AF112657), with six nucleotide changes at 72, 74, 423, 513, 549 and 627th bp found in the present isolate, causing an overall change of 4 amino acids. The 657 bp GST gene was cloned at BamH1 and HindIII restriction sites of the prokaryotic expression vector pPROEXHTb in frame with six histidine residues and expressed in E. coli DH5α. Recombinant protein was purified from the bacterial lysate under non-denaturing conditions by the process of sonication after lysozyme treatment and subjecting the soluble fraction of the bacterial lysate to Ni-NTA affinity chromatography. Western blotting with rabbit hyper-immune serum showed immuno-reactivity with 25 kDa recombinant GST. Recombinant protein detected F. gigantica experimental as well as field infection in buffaloes by dot-ELISA. However, cross-reactivity studies on Fasciola gigantica GST antigen are needed to evaluate the utility of this protein in the serodiagnosis of fasciolosis.Keywords: fasciola gigantic, fasciola hepatica, GST, RT- PCR
Procedia PDF Downloads 180721 Nutraceutical Characterization of Optimized Shatavari Asparagus racemosus Willd (Asparagaceae) Low Alcohol Nutra Beverage
Authors: Divya Choudhary, Hariprasad P., S. N. Naik
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This study examines a low-alcohol nutra-beverage made with shatavari, a plant commonly used in traditional medicine. During fermentation, the addition of a specific strain of yeast affected the beverage's properties, including its pH level, yeast count, ethanol content, and antioxidant, phenolic, and flavonoid levels. We also analyzed the beverage's storage and shelf life. Despite its bitter taste, the low alcohol content of the beverage made it enjoyable to drink and visually appealing. Our analysis showed that the optimal time for fermentation was between the 14th and 21st day when the beverage had ideal levels of sugar, organic acids, and vitamins. The final product contained fructose and citric acid but not succinic, pyruvic, lactic, or acetic acids. It also contained vitamins B2, B1, B12, and B9. During the shelf life analysis, we observed changes in the beverage's pH, TSS, and cfu levels, as well as its antioxidant activity. We also identified volatile (GC-MS) and non-volatile compounds (LC-MS/MS) in the fermented product, some of which were already present in the Shatavari root. The highest yield of product contained the maximum concentration of antioxidant compounds, which depended on both the pH and the microorganisms' physiological status. Overall, our study provides insight into the properties and potential health benefits of this Nutra-beverage.Keywords: antioxidants, fermentation, volatile compounds, acetonin, 1-butanol, non-volatile compounds, Shatavarin V, IX, kaempferol
Procedia PDF Downloads 59720 Esterification Reaction of Stearic Acid with Methanol Over Surface Functionalised PAN Fibrous Solid Acid Catalyst
Authors: Rawaz A. Ahmed, Katherine Huddersman
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High-lipid Fats, Oils and Grease (FOGs) from wastewater are underutilized despite their potential for conversion into valuable fuels; this work describes a surface-functionalized fibrous Polyacrylonitrile (PAN) mesh as a novel heterogeneous acid catalyst for the conversion of free fatty acids (FFAs), via a catalytic esterification process into biodiesel. The esterification of stearic acid (SA) with methanol was studied over an acidified PAN solid acid catalyst. Disappearance of the carboxylic acid (C=O) peak of the stearic acid at 1696 cm-1 in the FT-IR spectrum with the associated appearance of the ester (C=O) peak at 1739 cm-1 confirmed the production of the methyl stearate. This was further supported by 1H NMR spectra with the appearance of the ester (-CH₂OCOR) at 3.60-3.70 ppm. Quantitate analysis by GC-FID showed the catalyst has excellent activity with >95 % yield of methyl stearate (MS) at 90 ◦C after 3 h and a molar ratio of methanol to SA of 35:1. To date, to our best knowledge, there is no research in the literature on the esterification reaction for biodiesel production using a modified PAN mesh as a catalyst. It is noteworthy that this acidified PAN mesh catalyst showed comparable activity to conventional Brönsted acids, namely H₂SO₄ and p-TSA, as well as exhibiting higher activity than various other heterogeneous catalysts such as zeolites, ion-exchange resins and acid clay.Keywords: fats oil and greases (FOGs), free fatty acid, esterification reaction, methyl ester, PAN
Procedia PDF Downloads 228719 Storage Durations Affect the Physico-Chemical Characteristics of Physalis Minima L.
Authors: Norhanizan U., S. H. Ahmad, N. A. P. Abdullah, G. B. Saleh
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Physalis minima from the family of Solanaceae is one of the promising fruits which contains the high amount of vitamin C and other antioxidants as well. However, it is a perishable fruit where the deterioration process will commence if the fruits are not stored in proper conditions. There is not much work has been carried out to study the effects of storage durations on Physalis fruit. Therefore, this study was conducted to determine the effects of 0, 3, 6, and 9 days of storage on postharvest quality of Physalis minima fruits. Total of 120g of uniform sizes of fruits (2.3 to 2.5g) were used for each replication and the experiment was repeated thrice. The fruits were divided equally into four groups with each group labeled according to the days of storage. The fruits were then stored in the cool room for nine days with temperature maintain at 12 ° C. The fruits were analyzed for weight loss, firmness, color (L*, C* and hue angle), titratable acidity (TA), soluble solids concentrations (SSC), pH and ascorbic acids. Data were analyzed using analysis of variance and means was separated using least significant difference (LSD). The storage durations affect the quality characteristics of the fruits. On the day 9, the average of fruit weight loss and fruit firmness decreased about 21 and 24% respectively. The level of ascorbic acids and titrable acidity were also decreased while the soluble solids concentration increased during storage. Thus, in order to retain the quality of the fruits, it is recommended that the Physalis fruit can be stored only up to 6 days at 12 ° C.Keywords: fruit quality, Physalis minima, Solanaceae, storage durations
Procedia PDF Downloads 277718 Electrospun Fibers Made from Biopolymers (Cellulose Acetate/Chitosan) for Metals Recovery
Authors: Mauricio Gómez, Esmeralda López, Ian Becar, Jaime Pizarro, Paula A. Zapata
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A biodegradable material is developed with adsorptive capacity for metals ion for intended use in mining tailings mitigating the environmental impact with economic retribution, two types of fibers were elaborated by electrospinning: (1) a cellulose acetate (CA) matrix and (2) a cellulose acetate (CA)/chitosan (CH) matrix evaluating the effect of CH in CA on its physicochemical properties. Through diffuse reflectance infrared fourier transform spectroscopy (DRIFTS) the incorporation of chitosan in the matrix was identified, observing the band of the amino group at 1500 - 1600 [cm-1]. By scanning electron microscopy (SEM), Hg porosimetry, and CO2 isotherm at 273 [K], the intrafiber microporosity and interfiber macroporosity were identified, with an increase in the distribution of macropores for CA/CH fibers. In the tensile test, CH into the matrix produces a more ductile and tenacious behavior, where the % elongation at break increased by 33% with the other parameters constant. Thermal analysis by differential scanning calorimetry (DSC) and Thermogravimetric Analysis (TGA) showed that the incorporation of chitosan produces higher retention of water molecules due to the functional groups (amino groups (- NH3)), but there is a decrease in the specific heat and thermoplastic properties of the matrix since the glass transition temperature and softening temperature disappear. The effect of the optimum pH for CA and CA/CH fibers were studied in a batch system. In the adsorption kinetic study, the best isotherm model adapted to the experimental results corresponds to the Sips model and the kinetics corresponds to pseudo-second orderKeywords: environmental materials, wastewater treatment, electrospun fibers, biopolymers (cellulose acetate/chitosan), metals recovery
Procedia PDF Downloads 70717 Identification and Characterization of Polysaccharide Biosynthesis Protein (CAPD) of Enterococcus faecium
Authors: Liaqat Ali, Hubert E. Blum, Türkân Sakinc
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Enterococcus faecium is an emerging multidrug-resistant nosocomial pathogen increased dramatically worldwide and causing bacteremia, endocarditis, urinary tract and surgical site infections in immunocomprised patients. The capsular polysaccharides that contribute to pathogenesis through evasion of the host innate immune system are also involved in hindering leukocyte killing of enterococci. The gene cluster (enterococcal polysaccharide antigen) of E. faecalis encoding homologues of many genes involved in polysaccharide biosynthesis. We identified two putative loci with 22 kb and 19 kb which contained 11 genes encoding for glycosyltransferases (GTFs); this was confirmed by using genome comparison of already sequenced strains that has no homology to known capsule genes and the epa-locus. The polysaccharide-conjugate vaccines have rapidly emerged as a suitable strategy to combat different pathogenic bacteria, therefore, we investigated a polysaccharide biosynthesis CapD protein in E. faecium contains 336 amino acids and had putative function for N-linked glycosylation. The deletion/knock-out capD mutant was constructed and complemented by homologues recombination method and confirmed by using PCR and sequencing. For further characterization and functional analysis, in-vitro cell culture and in-vivo a mouse infection models were used. Our ΔcapD mutant shows a strong hydrophobicity and all strains exhibited biofilm production. Subsequently, the opsonic activity was tested in an opsonophagocytic assay which shows increased in mutant compared complemented and wild type strains but more than two fold decreased in colonization and adherence was seen on surface of uroepithelial cells. However, a significant higher bacterial colonialization was observed in capD mutant during animal bacteremia infection. Unlike other polysaccharides biosynthesis proteins, CapD does not seems to be a major virulence factor in enterococci but further experiments and attention is needed to clarify its function, exact mechanism and involvement in pathogenesis of enteroccocal nosocomial infections eventually to develop a vaccine/ or targeted therapy.Keywords: E. faecium, pathogenesis, polysaccharides, biofilm formation
Procedia PDF Downloads 323716 Development of Drug Delivery Systems for Endoplasmic Reticulum Amino Peptidases Modulators Using Electrospinning
Authors: Filipa Vasconcelos
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The administration of endoplasmic reticulum amino peptidases (ERAP1 or ERAP2) inhibitors can be used for therapeutic approaches against cancer and auto-immune diseases. However, one of the main shortcomings of drug delivery systems (DDS) is associated with the drug off-target distribution, which can lead to an increase in its side effects on the patient’s body. To overcome such limitations, the encapsulation of four representative compounds of ERAP inhibitors into Polycaprolactone (PCL), Polyvinyl-alcohol (PVA), crosslinked PVA, and PVA with nanoparticles (liposomes) electrospun fibrous meshes is proposed as a safe and controlled drug release system. The use of electrospun fibrous meshes as a DDS allows efficient solvent evaporation giving limited time to the encapsulated drug to recrystallize, continuous delivery of the drug while the fibers degrade, prevention of initial burst release (sustained release), tunable dosages, and the encapsulation of other agents. This is possible due to the fibers' small diameters and resemblance to the extracellular matrix (confirmed by scanning electron microscopy results), high specific surface area, and good mechanical strength/stability. Furthermore, release studies conducted on PCL, PVA, crosslinked PVA, and PVA with nanoparticles (liposomes) electrospun fibrous meshes with each of the ERAP compounds encapsulated demonstrated that they were capable of releasing >60%, 50%, 40%, and 45% of the total ERAP concentration, respectively. Fibrous meshes with ERAP_E compound encapsulated achieved higher released concentrations (75.65%, 62.41%, 56.05%, and 65.39%, respectively). Toxicity studies of fibrous meshes with encapsulated compounds are currently being accessed in vitro, as well as pharmacokinetics and dynamics studies. The last step includes the implantation of the drug-loaded fibrous meshes in vivo.Keywords: drug delivery, electrospinning, ERAP inhibitors, liposomes
Procedia PDF Downloads 95715 Submicron Laser-Induced Dot, Ripple and Wrinkle Structures and Their Applications
Authors: P. Slepicka, N. Slepickova Kasalkova, I. Michaljanicova, O. Nedela, Z. Kolska, V. Svorcik
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Polymers exposed to laser or plasma treatment or modified with different wet methods which enable the introduction of nanoparticles or biologically active species, such as amino-acids, may find many applications both as biocompatible or anti-bacterial materials or on the contrary, can be applied for a decrease in the number of cells on the treated surface which opens application in single cell units. For the experiments, two types of materials were chosen, a representative of non-biodegradable polymers, polyethersulphone (PES) and polyhydroxybutyrate (PHB) as biodegradable material. Exposure of solid substrate to laser well below the ablation threshold can lead to formation of various surface structures. The ripples have a period roughly comparable to the wavelength of the incident laser radiation, and their dimensions depend on many factors, such as chemical composition of the polymer substrate, laser wavelength and the angle of incidence. On the contrary, biopolymers may significantly change their surface roughness and thus influence cell compatibility. The focus was on the surface treatment of PES and PHB by pulse excimer KrF laser with wavelength of 248 nm. The changes of physicochemical properties, surface morphology, surface chemistry and ablation of exposed polymers were studied both for PES and PHB. Several analytical methods involving atomic force microscopy, gravimetry, scanning electron microscopy and others were used for the analysis of the treated surface. It was found that the combination of certain input parameters leads not only to the formation of optimal narrow pattern, but to the combination of a ripple and a wrinkle-like structure, which could be an optimal candidate for cell attachment. The interaction of different types of cells and their interactions with the laser exposed surface were studied. It was found that laser treatment contributes as a major factor for wettability/contact angle change. The combination of optimal laser energy and pulse number was used for the construction of a surface with an anti-cellular response. Due to the simple laser treatment, we were able to prepare a biopolymer surface with higher roughness and thus significantly influence the area of growth of different types of cells (U-2 OS cells).Keywords: cell response, excimer laser, polymer treatment, periodic pattern, surface morphology
Procedia PDF Downloads 229714 Light and Electron Study of Acrylamide–Induced Hypothalamic Changes
Authors: Keivan Jamshidi
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Distal swelling and eventual degeneration of axon in the CNS and PNS have been considered to be the characteristic neuropathological effects of acrylamide (ACR) neuropathy. This study was conducted to determine the neurotoxic effects of different doses of ACR (0.5, 5, 50, 100, and 500 mg/kg per day × 11days i. p.) on hypothalamus of rat using the de Olmos amino cupric-silver stain and electron microscopy. For this purpose 60 adult male rats (Wistar, approximately 250 g) were randomly assigned in 5 treatment groups as A, B, C, D, E) exposed to 0.5, 5, 50, 100, and 500 mg/kg per dayx11days i. p. and one control group as F received daily i. p. injections of 0.9% saline (3ml/kg). As indices of developing neurotoxicity, weight gain, gait scores and landing hindlimb foot splay were determined. After 11 days, two rats for silver stain, and two rats for EM were randomly selected; dissected and proper samples were collected from hypothalamus. Results did show no neurological behavior in groups A, B and F were observed in group C. Rats in groups D and E died within 1-2 hours due to sever toxemia. In histopathological studies based on de Olmos technique no argyrophilic neurons or processes were observed in stained sections obtained from hypothalamus of rats belong to groups A, B, and F while moderate to severe argyrophilic changes were observed in different nuclei and regions of stained sections obtained from hypothalamus of rats belong to group C. In ultra-structural studies some variations in the myelin sheet of injured axons including decompactation, interlaminar space formation, disruption of the laminar sheet, accumulation of neurofilaments, vacculation, and clumping inside the axolem, and finally complete disappearance of laminar sheet were observed.Keywords: acrylamide, hypothalamus, rat, de Olmos amino cupric, silver stain, electron microscopy
Procedia PDF Downloads 522713 Comparison of Existing Predictor and Development of Computational Method for S- Palmitoylation Site Identification in Arabidopsis Thaliana
Authors: Ayesha Sanjana Kawser Parsha
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S-acylation is an irreversible bond in which cysteine residues are linked to fatty acids palmitate (74%) or stearate (22%), either at the COOH or NH2 terminal, via a thioester linkage. There are several experimental methods that can be used to identify the S-palmitoylation site; however, since they require a lot of time, computational methods are becoming increasingly necessary. There aren't many predictors, however, that can locate S- palmitoylation sites in Arabidopsis Thaliana with sufficient accuracy. This research is based on the importance of building a better prediction tool. To identify the type of machine learning algorithm that predicts this site more accurately for the experimental dataset, several prediction tools were examined in this research, including the GPS PALM 6.0, pCysMod, GPS LIPID 1.0, CSS PALM 4.0, and NBA PALM. These analyses were conducted by constructing the receiver operating characteristics plot and the area under the curve score. An AI-driven deep learning-based prediction tool has been developed utilizing the analysis and three sequence-based input data, such as the amino acid composition, binary encoding profile, and autocorrelation features. The model was developed using five layers, two activation functions, associated parameters, and hyperparameters. The model was built using various combinations of features, and after training and validation, it performed better when all the features were present while using the experimental dataset for 8 and 10-fold cross-validations. While testing the model with unseen and new data, such as the GPS PALM 6.0 plant and pCysMod mouse, the model performed better, and the area under the curve score was near 1. It can be demonstrated that this model outperforms the prior tools in predicting the S- palmitoylation site in the experimental data set by comparing the area under curve score of 10-fold cross-validation of the new model with the established tools' area under curve score with their respective training sets. The objective of this study is to develop a prediction tool for Arabidopsis Thaliana that is more accurate than current tools, as measured by the area under the curve score. Plant food production and immunological treatment targets can both be managed by utilizing this method to forecast S- palmitoylation sites.Keywords: S- palmitoylation, ROC PLOT, area under the curve, cross- validation score
Procedia PDF Downloads 67712 Evaluation of Oxidative Changes in Soybean Oil During Shelf-Life by Physico-Chemical Methods and Headspace-Liquid Phase Microextraction (HS-LPME) Technique
Authors: Maryam Enteshari, Kooshan Nayebzadeh, Abdorreza Mohammadi
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In this study, the oxidative stability of soybean oil under different storage temperatures (4 and 25˚C) and during 6-month shelf-life was investigated by various analytical methods and headspace-liquid phase microextraction (HS-LPME) coupled to gas chromatography-mass spectrometry (GC-MS). Oxidation changes were monitored by analytical parameters consisted of acid value (AV), peroxide value (PV), p-Anisidine value (p-AV), thiobarbituric acid value (TBA), fatty acids profile, iodine value (IV), and oxidative stability index (OSI). In addition, concentrations of hexanal and heptanal as secondary volatile oxidation compounds were determined by HS-LPME/GC-MS technique. Rate of oxidation in soybean oil which stored at 25˚C was so higher. The AV, p-AV, and TBA were gradually increased during 6 months while the amount of unsaturated fatty acids, IV, and OSI decreased. Other parameters included concentrations of both hexanal and heptanal, and PV exhibited increasing trend during primitive months of storage; then, at the end of third and fourth months a sudden decrement was understood for the concentrations of hexanal and heptanal and the amount of PV, simultaneously. The latter parameters increased again until the end of shelf-time. As a result, the temperature and time were effective factors in oxidative stability of soybean oil. Also intensive correlations were found for soybean oil at 4 ˚C between AV and TBA (r2=0.96), PV and p-AV (r2=0.9), IV and TBA (-r2=0.9), and for soybean oil stored at 4˚C between p-AV and TBA (r2=0.99).Keywords: headspace-liquid phase microextraction, oxidation, shelf-life, soybean oil
Procedia PDF Downloads 396711 Effects of Vegetable Oils Supplementation on in Vitro Rumen Fermentation and Methane Production in Buffaloes
Authors: Avijit Dey, Shyam S. Paul, Satbir S. Dahiya, Balbir S. Punia, Luciano A. Gonzalez
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Methane emitted from ruminant livestock not only reduces the efficiency of feed energy utilization but also contributes to global warming. Vegetable oils, a source of poly unsaturated fatty acids, have potential to reduce methane production and increase conjugated linoleic acid in the rumen. However, characteristics of oils, level of inclusion and composition of basal diet influences their efficacy. Therefore, this study was aimed to investigate the effects of sunflower (SFL) and cottonseed (CSL) oils on methanogenesis, volatile fatty acids composition and feed fermentation pattern by in vitro gas production (IVGP) test. Four concentrations (0, 0.1, 0.2 and 0.4ml /30ml buffered rumen fluid) of each oil were used. Fresh rumen fluid was collected before morning feeding from two rumen cannulated buffalo steers fed a mixed ration. In vitro incubation was carried out with sorghum hay (200 ± 5 mg) as substrate in 100 ml calibrated glass syringes following standard IVGP protocol. After 24h incubation, gas production was recorded by displacement of piston. Methane in the gas phase and volatile fatty acids in the fermentation medium were estimated by gas chromatography. Addition of oils resulted in increase (p<0.05) in total gas production and decrease (p<0.05) in methane production, irrespective of type and concentration. Although the increase in gas production was similar, methane production (ml/g DM) and its concentration (%) in head space gas was lower (p< 0.01) in CSL than in SFL at corresponding doses. Linear decrease (p<0.001) in degradability of DM was evident with increasing doses of oils (0.2ml onwards). However, these effects were more pronounced with SFL. Acetate production tended to decrease but propionate and butyrate production increased (p<0.05) with addition of oils, irrespective of type and doses. The ratio of acetate to propionate was reduced (p<0.01) with addition of oils but no difference between the oils was noted. It is concluded that both the oils can reduce methane production. However, feed degradability was also affected with higher doses. Cotton seed oil in small dose (0.1ml/30 ml buffered rumen fluid) exerted greater inhibitory effects on methane production without impeding dry matter degradability. Further in vivo studies need to be carried out for their practical application in animal ration.Keywords: buffalo, methanogenesis, rumen fermentation, vegetable oils
Procedia PDF Downloads 396710 Mechanisms of Metals Stabilization in the Soil by Biochar Material as Affected by the Low Molecular Weight Organic Acids
Authors: Md. Shoffikul Islam, Hongqing Hu
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Immobilizing trace elements by reducing their mobility and bioavailability through amendment application, especially biochar (BC), is a cost-effective and efficient method to address their toxicity in the soil environment. However, the low molecular weight organic acids (LMWOAs) in the rhizosphere could affect BC's efficiency to immobilize trace metals as the LMWOAs could either mobilize or fix metals in the soils. Therefore, understanding the BC's and LMWOAs' interaction mechanisms on metals stabilization in the rhizosphere is crucial. The present study examined the impact of BC derived from rice husk, tartaric acid (TA), and oxalic acid (OA), and the combination of BC and TA/OA on the changes of cadmium (Cd), lead (Pb), and zinc (Zn) among their geochemical forms through incubation experiment. The changes of zeta potential and X-ray diffraction (XRD) pattern of BC and BC-amended soils to investigate the probable mechanisms of trace elements' immobilization by BC under the attacks of TA and OA were also examined. The rice husk BC at 5% (w/w) was mixed with the air-dry soil (an Anthrosols) contaminated with Cd, Pb, and Zn in the plastic pot. The TA and OA each at 2, 5, 10, and 20 mM kg-1 (w/v) were added separately into the pot. All the ingredients were mixed thoroughly with the soil. A control (CK) treatment was also prepared without BC, TA, and OA addition. After 7, 15, and 60 days of incubation with 60% (w/v) moisture level at 25 °C, the incubated soils were determined for pH and EC and were sequentially extracted to assess the metals' transformation in soil. The electronegative charges and XRD peaks of BC and BC-amended soils were also measured. The BC, low level of TA (2 mM kg-1 soil), and BC plus the low concentration of TA (BC-TA2) addition considerably declined the acid-soluble Cd, Pb, and Zn in which BC-TA2 was found to be the most effective treatment. The trends were reversed concerning the high levels of TA (>5-20 mM kg-1 soil), all levels of OA (2-20 mM kg-1 soil), and the BC plus high levels of TA/OA treatments. BC-TA2 changed the highest amounts of acid-soluble and reducible metals to the oxidizable and residual fractions with time. The most increased electronegative charges of BC-TA2 indicate its (BC-TA2) highest metals' immobilizing efficiency, probably through metals adsorption and fixation with the negative charge sites. The XRD study revealed the presence of P, O, CO32-, and Cl1- in BC, which might be responsible for the precipitation of CdCO3, pyromorphite, and hopeite concerning Cd, Pb, and Zn immobilization, respectively. The findings demonstrated that the low level of TA increased metals immobilization, while the high levels of TA and all levels of OA enhanced their mobilization. The BC-TA2 was the best treatment in stabilizing metals in soil.Keywords: biochar, immobilization, low molecular weight organic acids, trace elements contaminated soil
Procedia PDF Downloads 74709 Anti-Inflammatory Effect of Omega-3 Fish-Oil Supplements: Eicosapentaenoic Acid and Docosahexaenoic Acid in Early-Stage Tumors
Authors: Corina Muscurel, Irina Stoian, Laura Gaman, Valeriu Atanasiu
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Chronic inflammation predisposes cells to neoplastic transformation and is associated with angiogenesis. Omega-3 polyunsaturated fatty acids (n-3 PUFAs) give rise to anti-inflammatory metabolites and decrease some inflammatory cytokines. The aim of the study was to analyze the effect of n-3 PUFAs intake on patients with tumors in early-stage (without regional or distant metastasis). There were two groups of patients: one group with colon tumors and one group with lung tumors. All patients took for 60 days daily supplements from fish-oil containing 600 mg eicosapentaenoic acid and 400 mg docosahexaenoic acid. The plasma markers were evaluated before and after PUFAs intake: ceruloplasmin (using p-phenylenediamine oxidase method), plasma total thiol groups (using dithiobis-nitrobenzoic acid method) and CEA (carcinoembryonic antigen using electrochemiluminescent immunoassay). The results reflect ceruloplasmin decrease (p < 0.05), plasma total thiol groups increase (not statistically significant) and CEA decrease (p < 0.05) after n-3 PUFAs intake. Conclusions: n-3 PUFAs intake is favorable in premalignant lesions or in early tumor stage and dietary fish-oil has anti-inflammatory effects and can contribute to reduce cancer progression.Keywords: cancer, fish-oil, inflammation, n-3 polyunsaturated fatty acids
Procedia PDF Downloads 129708 Evaluation of Functional Properties of Protein Hydrolysate from the Fresh Water Mussel Lamellidens marginalis for Nutraceutical Therapy
Authors: Jana Chakrabarti, Madhushrita Das, Ankhi Haldar, Roshni Chatterjee, Tanmoy Dey, Pubali Dhar
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High incidences of Protein Energy Malnutrition as a consequence of low protein intake are quite prevalent among the children in developing countries. Thus prevention of under-nutrition has emerged as a critical challenge to India’s developmental Planners in recent times. Increase in population over the last decade has led to greater pressure on the existing animal protein sources. But these resources are currently declining due to persistent drought, diseases, natural disasters, high-cost of feed, and low productivity of local breeds and this decline in productivity is most evident in some developing countries. So the need of the hour is to search for efficient utilization of unconventional low-cost animal protein resources. Molluscs, as a group is regarded as under-exploited source of health-benefit molecules. Bivalve is the second largest class of phylum Mollusca. Annual harvests of bivalves for human consumption represent about 5% by weight of the total world harvest of aquatic resources. The freshwater mussel Lamellidens marginalis is widely distributed in ponds and large bodies of perennial waters in the Indian sub-continent and well accepted as food all over India. Moreover, ethno-medicinal uses of the flesh of Lamellidens among the rural people to treat hypertension have been documented. Present investigation thus attempts to evaluate the potential of Lamellidens marginalis as functional food. Mussels were collected from freshwater ponds and brought to the laboratory two days before experimentation for acclimatization in laboratory conditions. Shells were removed and fleshes were preserved at- 20oC until analysis. Tissue homogenate was prepared for proximate studies. Fatty acids and amino acids composition were analyzed. Vitamins, Minerals and Heavy metal contents were also studied. Mussel Protein hydrolysate was prepared using Alcalase 2.4 L and degree of hydrolysis was evaluated to analyze its Functional properties. Ferric Reducing Antioxidant Power (FRAP) and DPPH Antioxidant assays were performed. Anti-hypertensive property was evaluated by measuring Angiotensin Converting Enzyme (ACE) inhibition assay. Proximate analysis indicates that mussel meat contains moderate amount of protein (8.30±0.67%), carbohydrate (8.01±0.38%) and reducing sugar (4.75±0.07%), but less amount of fat (1.02±0.20%). Moisture content is quite high but ash content is very low. Phospholipid content is significantly high (19.43 %). Lipid constitutes, substantial amount of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) which have proven prophylactic values. Trace elements are found present in substantial amount. Comparative study of proximate nutrients between Labeo rohita, Lamellidens and cow’s milk indicates that mussel meat can be used as complementary food source. Functionality analyses of protein hydrolysate show increase in Fat absorption, Emulsification, Foaming capacity and Protein solubility. Progressive anti-oxidant and anti-hypertensive properties have also been documented. Lamellidens marginalis can thus be regarded as a functional food source as this may combine effectively with other food components for providing essential elements to the body. Moreover, mussel protein hydrolysate provides opportunities for utilizing it in various food formulations and pharmaceuticals. The observations presented herein should be viewed as a prelude to what future holds.Keywords: functional food, functional properties, Lamellidens marginalis, protein hydrolysate
Procedia PDF Downloads 415707 Development of Site-Specific Colonic Drug Delivery System (Nanoparticles) of Chitosan Coated with pH Sensitive Polymer for the Management of Colonic Inflammation
Authors: Pooja Mongia Raj, Rakesh Raj, Alpana Ram
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Background: The use of multiparticulate drug delivery systems in preference to single unit dosage forms for colon targeting purposes dates back to 1985 when Hardy and co-workers showed that multiparticulate systems enabled the drug to reach the colon quickly and were retained in the ascending colon for a relatively long period of time. Methods: Site-specific colonic drug delivery system (nanoparticles) of 5-ASA were prepared and coated with pH sensitive polymer. Chitosan nanoparticles (CTNP) bearing 5-Amino salicylic acid (5-ASA) were prepared, by ionotropic gelation method. Nanoparticulate dosage form consisting of a hydrophobic core enteric coated with pH-dependent polymer Eudragit S-100 by solvent evaporation method, for the effective delivery of drug to the colon for treatment of ulcerative colitis. Results: The mean diameter of CTNP and ECTNP formulations were 159 and 661 nm, respectively. Also optimum value of polydispersity index was found to be 0.249 [count rate (kcps) was 251.2] and 0.170 [count rate (kcps) was 173.9] was obtained for both the formulations respectively. Conclusion: CTNP and Eudragit chitosan nanoparticles (ECTNP) was characterized for shape and surface morphology by scanning electron microscopy (SEM) appeared to be spherical in shape. The in vitro drug release was investigated using USP dissolution test apparatus in different simulated GIT fluids showed promising release. In vivo experiments are in further proceeding for fruitful results.Keywords: colon targeting, nanoparticles, polymer, 5-amino salicylic acid, edragit
Procedia PDF Downloads 489706 Functionality and Application of Rice Bran Protein Hydrolysates in Oil in Water Emulsions: Their Stabilities to Environmental Stresses
Authors: R. Charoen, S. Tipkanon, W. Savedboworn, N. Phonsatta, A. Panya
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Rice bran protein hydrolysates (RBPH) were prepared from defatted rice bran of two different Thai rice cultivars (Plai-Ngahm-Prachinburi; PNP and Khao Dok Mali 105; KDM105) using an enzymatic method. This research aimed to optimize enzyme-assisted protein extraction. In addition, the functional properties of RBPH and their stabilities to environmental stresses including pH (3 to 8), ionic strength (0 mM to 500 mM) and the thermal treatment (30 °C to 90 °C) were investigated. Results showed that enzymatic process for protein extraction of defatted rice bran was as follows: enzyme concentration 0.075 g/ 5 g of protein, extraction temperature 50 °C and extraction time 4 h. The obtained protein hydrolysate powders had a degree of hydrolysis (%) of 21.05% in PNP and 19.92% in KDM105. The solubility of protein hydrolysates at pH 4-6 was ranged from 27.28-38.57% and 27.60-43.00% in PNP and KDM105, respectively. In general, antioxidant activities indicated by total phenolic content, FRAP, ferrous ion-chelating (FIC), and 2,2’-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) of KDM105 had higher than PNP. In terms of functional properties, the emulsifying activity index (EAI) was was 8.78 m²/g protein in KDM105, whereas PNP was 5.05 m²/g protein. The foaming capacity at 5 minutes (%) was 47.33 and 52.98 in PNP and KDM105, respectively. Glutamine, Alanine, Valine, and Leucine are the major amino acid in protein hydrolysates where the total amino acid of KDM105 gave higher than PNP. Furthermore, we investigated environmental stresses on the stability of 5% oil in water emulsion (5% oil, 10 mM citrate buffer) stabilized by RBPH (3.5%). The droplet diameter of emulsion stabilized by KDM105 was smaller (d < 250 nm) than produced by PNP. For environmental stresses, RBPH stabilized emulsions were stable at pH around 3 and 5-6, at high salt (< 400 mM, pH 7) and at temperatures range between 30-50°C.Keywords: functional properties, oil in water emulsion, protein hydrolysates, rice bran protein
Procedia PDF Downloads 206705 Hyper-Production of Lysine through Fermentation and Its Biological Evaluation on Broiler Chicks
Authors: Shagufta Gulraiz, Abu Saeed Hashmi, Muhammad Mohsin Javed
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Lysine required for poultry feed is imported in Pakistan to fulfil the desired dietary needs. Present study was designed to produce maximum lysine by utilizing cheap sources to save the foreign exchange. To achieve the goal of lysine production through fermentation, large scale production of lysine was carried out in 7.5 L stirred glass vessel fermenter with wild and mutant Brevibacterium flavum (B. flavum) using all pre-optimized conditions. The identification of produced lysine was carried out by TLC and amino acid analyzer. Toxicity evaluation of produced lysine was performed before feeding to broiler chicks. During biological trial concentrated fermented broth having 8% lysine was used in poultry rations as a source of Lysine for test birds. Fermenter scale studies showed that the maximum lysine (20.8 g/L) was produced at 250 rpm, 1.5 vvm aeration, 6.0% inoculum under controlled pH conditions after 56 h of fermentation with wild culture but mutant (BFENU2) gave maximum yield of lysine 36.3 g/L under optimized condition after 48 h. Amino acid profiling showed 1.826% Lysine in fermented broth by wild B. flavum and 2.644% by mutant strain (BFENU2). Toxicity evaluation report showed that the produced lysine is safe for consumption by broilers. Biological evaluation results showed that produced lysine was equally good as commercial lysine in terms of weight gain, feed intake and feed conversion ratio. A cheap and practical bioprocess of Lysine production was concluded, that can be exploited commercially in Pakistan to save foreign exchange.Keywords: lysine, fermentation, broiler chicks, biological evaluation
Procedia PDF Downloads 543704 Repeatable Surface Enhanced Raman Spectroscopy Substrates from SERSitive for Wide Range of Chemical and Biological Substances
Authors: Monika Ksiezopolska-Gocalska, Pawel Albrycht, Robert Holyst
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Surface Enhanced Raman Spectroscopy (SERS) is a technique used to analyze very low concentrations of substances in solutions, even in aqueous solutions - which is its advantage over IR. This technique can be used in the pharmacy (to check the purity of products); forensics (whether at a crime scene there were any illegal substances); or medicine (serving as a medical test) and lots more. Due to the high potential of this technique, its increasing popularity in analytical laboratories, and simultaneously - the absence of appropriate platforms enhancing the SERS signal (crucial to observe the Raman effect at low analyte concentration in solutions (1 ppm)), we decided to invent our own SERS platforms. As an enhancing layer, we have chosen gold and silver nanoparticles, because these two have the best SERS properties, and each has an affinity for the other kind of particles, which increases the range of research capabilities. The next step was to commercialize them, which resulted in the creation of the company ‘SERSitive.eu’ focusing on production of highly sensitive (Ef = 10⁵ – 10⁶), homogeneous and reproducible (70 - 80%) substrates. SERStive SERS substrates are made using the electrodeposition of silver or silver-gold nanoparticles technique. Thanks to a very detailed analysis of data based on studies optimizing such parameters as deposition time, temperature of the reaction solution, applied potential, used reducer, or reagent concentrations using a standardized compound - p-mercaptobenzoic acid (PMBA) at a concentration of 10⁻⁶ M, we have developed a high-performance process for depositing precious metal nanoparticles on the surface of ITO glass. In order to check a quality of the SERSitive platforms, we examined the wide range of the chemical compounds and the biological substances. Apart from analytes that have great affinity to the metal surfaces (e.g. PMBA) we obtained very good results for those fitting less the SERS measurements. Successfully we received intensive, and what’s more important - very repetitive spectra for; amino acids (phenyloalanine, 10⁻³ M), drugs (amphetamine, 10⁻⁴ M), designer drugs (cathinone derivatives, 10⁻³ M), medicines and ending with bacteria (Listeria, Salmonella, Escherichia coli) and fungi.Keywords: nanoparticles, Raman spectroscopy, SERS, SERS applications, SERS substrates, SERSitive
Procedia PDF Downloads 146703 Genomic and Evolutionary Diversity of Long Terminal Repeat (LTR) Retrotransposons in Date Palm (Phoenix dactylifera)
Authors: Faisal Nouroz, Mukaramin Mukaramin
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Of the transposable elements (TEs), the retrotransposons are the most copious elements identified from many sequenced genomes. They have played a major role in genome evolution, rearrangement, and expansions based on their copy and paste mode of proliferation. They are further divided into LTR and Non-LTR retrotransposons. The purpose of the current study was to identify the LTR REs in sequenced Phoenix dactylifera genome and to study their structural diversity. A total of 150 P. dactylifera BAC sequences with > 60kb sizes were randomly retrieved from National Center for Biotechnology Information (NCBI) database and screened for the presence of LTR retrotransposons. Seven bacterial artificial chromosomes (BAC) sequences showed full-length LTR Retrotransposons with 4 Copia and 3 Gypsy families having variable copy numbers in respective families. Reverse transcriptase (RT) domain was found as the most conserved domain among Copia and Gypsy superfamilies and was used to deduce evolutionary analysis. The amino acid residues among various RT sequences showed variability in their percentages indicating post divergence evolution. Amino acid Leucine was found in highest proportions followed by Lysine, while Methionine and Tryptophan were in lowest percentages. The phylogenetic analysis based on RT domains confirmed that although having most conserved RT regions, several evolutionary events occurred causing nucleotide polymorphisms and hence clustering of Gypsy and Copia superfamilies into their respective lineages. The study will be helpful in identification and annotation of these elements in other species and genera and their distribution patterns on chromosomes by fluorescent in situ hybridization techniques.Keywords: transposable elements, Phoenix dactylifera, retrotransposons, phylogenetic analysis
Procedia PDF Downloads 125702 Enhancement of Morphogenetic Potential to Obtain Elite Varities of Sauropus androgynous (L.) Merr. through Somatic Embryogenesis
Authors: S. Padma, D. H. Tejavathi
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Somatic embryogenesis is a remarkable illustration of the dictum of plant totipotency where developmental reconstruction of somatic cells takes place towards the embryogenic pathway. It recapitulates the morphological and developmental process that occurs in zygotic embryogenesis. S. androgynous commonly called as multivitamin plant. The leaves are consumed as green leafy vegetable by the Southeast Asian communities due to their rich nutritional profile. Despite being a good nutritional vegetable with proteins, vitamins, minerals, amino acids, it is warned for excessive intake due to the presence of alkoloid called papaverine. Papaverine at higher concentrations is toxic and leads to a syndrome called Bronchiolitis Obliterans. In the present study, morphogenetic potential of shoot tip, leaf and nodal explants of Sauropus androgynous was investigated to develop and enhance the reliable plant regeneration protocol via somatic embryogenesis. Somatic embryos were derived directly from the embryogenic callus derived from shoot tip, node and leaf cultures on Phillips and Collins (L2) medium supplemented with NAA at various concentrations ranging from 5.3 µM/l to 26.85 µM/l within two months of inoculation. Thus obtained embryos were sub cultured to modified L2 media supplemented with increased vitamin level for the further growth. Somatic embryos with well-developed cotyledons were transferred to normal and modified L2 basal medium for conversion. The plantlets thus obtained were subjected to brief acclimatization before transferring them to land. About 95% of survival rate was recorded. The augmentation process of culturing various explants through somatic embryogenesis using synthetic medium with various plant growth regulators under controlled conditions have aggrandized the commercial production of Sauropus making it easily available over the conventional propagation methods. In addition, regeneration process through somatic embryogenesis has ameliorated the development of desired character in Sauropus with low papaverine content thereby providing a valuable resource to the food and pharmaceutical industry. Based on this research, plant tissue culture techniques have shown promise for economical and convenient application in Sauropus androgynous breeding.Keywords: L2 medium, multivitamin plant, NAA, papaverine
Procedia PDF Downloads 203701 Comparison of Bioactive Compound Content in Egg Yolk Oil Extracted from Eggs Obtained from Different Laying Hen Housing Systems
Authors: Aleksandrs Kovalcuks
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Egg yolk oil is a natural source of bioactive compounds such as unsaturated fatty acids, oil soluble vitamins, pigments and others. Bioactive compound content in egg yolk oil depends from its content in eggs, from which oil was extracted. Many studies show that bioactive compound content in egg is correlated to the content of these compounds in hen feed, but there is also an opinion that hen housing systems also have influence on egg chemical content. The aim of this study was to determine which factor, laying hen housing system or hen diet, has a primary influence on bioactive compound content in egg yolk oil. The egg yolk oil was extracted from eggs obtained from 4 different hen housing systems: cage, barn and two groups of free range. All hens were fed with commercially produced compound feed except one group of free range hens which get free diet – pastured hens. Extracted egg yolk oils were analyzed for fatty acids, oil soluble vitamins and β-carotene content. α-tocopherol, ergocalcipherol and polyunsaturated fatty acid content in egg yolk oil was higher from eggs obtained from all housing systems where hens were fed with commercial compound feed. β-carotene and retinol content in egg yolk oils from free range free diet eggs was significantly (p>0.05) higher that from other eggs because hens have access to green forage. Hen physical activity in free range housing systems decreases content of some bioactive compound in egg yolk oil.Keywords: egg yolk oil, vitamins, caged eggs, free range
Procedia PDF Downloads 456700 Identification and Molecular Profiling of A Family I Cystatin Homologue from Sebastes schlegeli Deciphering Its Putative Role in Host Immunity
Authors: Don Anushka Sandaruwan Elvitigala, P. D. S. U. Wickramasinghe, Jehee Lee
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Cystatins are a large superfamily of proteins which act as reversible inhibitors of cysteine proteases. Papain proteases and cysteine cathepsins are predominant substrates of cystatins. Cystatin superfamily can be further clustered into three groups as Stefins, Cystatins, and Kininogens. Among them, stefines are also known as family 1 cystatins which harbors cystatin Bs and cystatin As. In this study, a homologue of family one cystatins more close to cystatin Bs was identified from Korean black rockfish (Sebastes schlegeli) using a prior constructed cDNA (complementary deoxyribonucleic acid) database and designated as RfCyt1. The full-length cDNA of RfCyt1 consisted of 573 bp, with a coding region of 294 bp. It comprised a 5´-untranslated region (UTR) of 55 bp, and 3´-UTR of 263 bp. The coding sequence encodes a polypeptide consisting of 97 amino acids with a predicted molecular weight of 11kDa and theoretical isoelectric point of 6.3. The RfCyt1 shared homology with other teleosts and vertebrate species and consisted conserved features of cystatin family signature including single cystatin-like domain, cysteine protease inhibitory signature of pentapeptide (QXVXG) consensus sequence and N-terminal two conserved neighboring glycine (⁸GG⁹) residues. As expected, phylogenetic reconstruction developed using the neighbor-joining method showed that RfCyt1 is clustered with the cystatin family 1 members, in which more closely with its teleostan orthologues. An SYBR Green qPCR (quantitative polymerase chain reaction) assay was performed to quantify the RfCytB transcripts in different tissues in healthy and immune stimulated fish. RfCyt1 was ubiquitously expressed in all tissue types of healthy animals with gill and spleen being the highest. Temporal expression of RfCyt1 displayed significant up-regulation upon infection with Aeromonas salmonicida. Recombinantly expressed RfCyt1 showed concentration-dependent papain inhibitory activity. Collectively these findings evidence for detectable protease inhibitory and immunity relevant roles of RfCyt1 in Sebastes schlegeli.Keywords: Sebastes schlegeli, family 1 cystatin, immune stimulation, expressional modulation
Procedia PDF Downloads 128699 Characterization on Molecular Weight of Polyamic Acids Using GPC Coupled with Multiple Detectors
Authors: Mei Hong, Wei Liu, Xuemin Dai, Yanxiong Pan, Xiangling Ji
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Polyamic acid (PAA) is the precursor of polyimide (PI) prepared by a two-step method, its molecular weight and molecular weight distribution not only play an important role during the preparation and processing, but also influence the final performance of PI. However, precise characterization on molecular weight of PAA is still a challenge because of the existence of very complicated interactions in the solution system, including the electrostatic interaction, hydrogen bond interaction, dipole-dipole interaction, etc. Thus, it is necessary to establisha suitable strategy which can completely suppress these complex effects and get reasonable data on molecular weight. Herein, the gel permeation chromatography (GPC) coupled with differential refractive index (RI) and multi-angle laser light scattering (MALLS) detectors were applied to measure the molecular weight of (6FDA-DMB) PAA using different mobile phases, LiBr/DMF, LiBr/H3PO4/THF/DMF, LiBr/HAc/THF/DMF, and LiBr/HAc/DMF, respectively. It was found that combination of LiBr with HAc can shield the above-mentioned complex interactions and is more conducive to the separation of PAA than only addition of LiBr in DMF. LiBr/HAc/DMF was employed for the first time as a mild mobile phase to effectively separate PAA and determine its molecular weight. After a series of conditional experiments, 0.02M LiBr/0.2M HAc/DMF was fixed as an optimized mobile phase to measure the relative and absolute molecular weights of (6FDA-DMB) PAA prepared, and the obtained Mw from GPC-MALLS and GPC-RI were 35,300 g/mol and 125,000 g/mol, respectively. Particularly, such a mobile phase is also applicable to other PAA samples with different structures, and the final results on molecular weight are also reproducible.Keywords: Polyamic acids, Polyelectrolyte effects, Gel permeation chromatography, Mobile phase, Molecular weight
Procedia PDF Downloads 49698 Synthesis and Preparation of Carbon Ferromagnetic Nanocontainers for Cancer Therapy
Authors: L. Szymanski, Z. Kolacinski, Z. Kamiński, G. Raniszewski, J. Fraczyk, L. Pietrzak
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In the article the development and demonstration of method and the model device for hyperthermic selective destruction of cancer cells are presented. This method was based on the synthesis and functionalization of carbon nanotubes serving as ferromagnetic material nano containers. Methodology of the production carbon - ferromagnetic nanocontainers includes: the synthesis of carbon nanotubes, chemical and physical characterization, increasing the content of ferromagnetic material and biochemical functionalization involving the attachment of the key addresses. Biochemical functionalization of ferromagnetic nanocontainers is necessary in order to increase the binding selectively with receptors presented on the surface of tumour cells. Multi-step modification procedure was finally used to attach folic acid on the surface of ferromagnetic nanocontainers. Folic acid is ligand of folate receptors which is overexpresion in tumor cells. The presence of ligand should ensure the specificity of the interaction between ferromagnetic nanocontainers and tumor cells. The chemical functionalization contains several step: oxidation reaction, transformation of carboxyl groups into more reactive ester or amide groups, incorporation of spacer molecule (linker), attaching folic acid. Activation of carboxylic groups was prepared with triazine coupling reagent (preparation of superactive ester attached on the nanocontainers). The spacer molecules were designed and synthesized. In order to ensure biocompatibillity of linkers they were built from amino acids or peptides. Spacer molecules were synthesized using the SPPS method. Synthesis was performed on 2-Chlorotrityl resin. The linker important feature is its length. Due to that fact synthesis of peptide linkers containing from 2 to 4 -Ala- residues was carried out. Independent synthesis of the conjugate of foilic acid with 6-aminocaproic acid was made. Final step of synthesis was connecting conjugat with spacer molecules and attaching it on the ferromagnetic nanocontainer surface. This article contains also information about special CVD and microvave plasma system to produce nanotubes and ferromagnetic nanocontainers. The first tests in the device for hyperthermal RF generator will be presented. The frequency of RF generator was in the ranges from 10 to 14Mhz and from 265 to 621kHz.Keywords: synthesis of carbon nanotubes, hyperthermia, ligands, carbon nanotubes
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