Search results for: pesticide residues

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

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Authors: Huanru Wang, Jianzhun Jiang

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At present, the preparation of the catalyst by carbon carrier is one of the improvement directions of the C₂ pre-hydrogenation catalyst. Carbon materials can be prepared from coal direct liquefaction residues, coconut shells, biomass, etc., and the pore structure of carbon carrier materials can be adjusted through the preparation process; at high temperatures, the carbon carrier itself also shows certain catalytic activity. Therefore, this paper mainly selected typical activated carbon and coconut shell carbon as carbon carrier materials, studied their microstructure and surface properties, prepared a series of carbon-based catalysts loaded with Pd, and investigated the effects of the content of promoter Ag and the concentration of reductant on the structure and performance of the catalyst and its catalytic performance for the pre hydrogenation of C₂. In this paper, the carbon supports from two sources and the catalysts prepared by them were characterized in detail. The results showed that the morphology and structure of different supports and the performance of the catalysts prepared were also obviously different. The catalyst supported on coconut shell carbon has a small specific surface area and large pore diameter. The catalyst supported on activated carbon has a large specific surface area and rich pore structure. The active carbon support is mainly a mixture of amorphous graphite and microcrystalline graphite. For the catalyst prepared with coconut shell carbon as the carrier, the sample is very uneven, and its specific surface area and pore volume are irregular. Compared with coconut shell carbon, activated carbon is more suitable as the carrier of the C₂ hydrogenation catalyst. The conversion of acetylene, methyl acetylene, and butadiene decreased, and the ethylene selectivity increased after Ag was added to the supported Pd catalyst. When the amount of promoter Ag is 0.01-0.015%, the catalyst has relatively good catalytic performance. Ag and Pd form an alloying effect, thus reducing the effective demand for Ag. The Pd Ag ratio is the key factor affecting the catalytic performance. When the addition amount of Ag is 0.01-0.015%, the dispersion of Pd on the carbon support surface can be significantly improved, and the size of active particles can be reduced. The Pd Ag ratio is the main factor in improving the selectivity of the catalyst. When the additional amount of sodium formate is 1%, the catalyst prepared has both high acetylene conversion and high ethylene selectivity.

Keywords: C₂ hydrogenation, activated carbon, Ag promoter, Pd catalysts

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Authors: Timine Suoware, Sylvester Edelugo, Charles Amgbari

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The susceptibility of natural fibre polymer composites to flame has necessitated research to improve and develop flame retardant (FR) to delay the escape of combustible volatiles. Previous approaches relied mostly on FR such as aluminium tri-hydroxide (ATH) and ammonium polyphosphate (APP) to improve fire performances of wood sawdust polymer composites (WSPC) with emphasis on non-structural building applications. In this paper, APP was modified with gum Arabic powder (GAP) and then hybridized with ATH at 0, 12 and 18% loading ratio to form new FR species; WSPC12%APP-GAP and WSPC18%ATH/APP-GAP. The FR species were incorporated in wood sawdust waste reinforced in polyester resin to form panels for fireproof doors. The panels were produced using hand lay compression moulding technique and cured at room temperature. Specimen cut from panels were then tested for tensile strength (TS), flexural strength (FS) and impact strength (IS) using universal testing machine and impact tester; thermal stability using (TGA/DSC 1: Metler Toledo); time-to-ignition (Tig), heat release rates (HRR); peak HRR (HRRp), average HRR (HRRavg), total HRR (THR), peak mass loss rate (MLRp), average smoke production rate (SPRavg) and carbon monoxide production (COP ) were obtained using the cone calorimeter apparatus. From the mechanical properties obtained, improvements of IS for the panels were not noticeable whereas TS and FS for WSPC12%APP-GAP respectively stood at 12.44 MPa and 85.58 MPa more than those without FR (WSPC0%). For WSC18%ATH/APP-GAP TS and FS respectively stood at 16.45 MPa and 50.49 MPa more compared to (WSPC0%). From the thermal analysis, the panels did not exhibit any significant change as early degradation was observed. At 900 OC, the char residues improved by 15% for WSPC12%APP-GAP and 19% for WSPC18%ATH/APP-GAP more than (WSC0%) at 5%, confirming the APP-GAP to be a good FR. At 50 kW/m2 heat flux (HF), WSPC12%APP-GAP improved better the fire behaviour of the panels when compared to WSC0% as follows; Tig = 46 s, HRRp = 56.1 kW/2, HRRavg = 32.8 kW/m2, THR = 66.6 MJ/m2, MLRp = 0.103 g/s, TSR = 0.04 m2/s and COP = 0.051 kg/kg. These were respectively more than WSC0%. It can be concluded that the new concept of modifying FR with GAP in WSC could meet the requirement of a fireproof door for building applications.

Keywords: composite, flame retardant, wood sawdust, fireproof doors

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Authors: Lali Shanshiashvili, Marika Chikviladze, Nino Mamulashvili, Maia Sepashvili, Nana Narmania, David Mikeladze

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Purpose: During demyelinating inflammatory diseases and after the damage of the myelin sheet, myelin-derived proteins, including myelin basic protein (MBP), are secreted into the extracellular space. MBP shows extensive post-translational modifications, including the deimination of arginine residues. Deiminated MBP is structurally less ordered, susceptible to proteolytic attack, and more immunogenic than the unmodified one. It is hypothesized that MBP could change the inflammatory response in astrocytes. Methods: MBP was isolated and purified from bovine brain white matter. Primary astrocyte cultures were prepared from whole brains of 2-day-old Wistar rats. For evaluation of glutamate uptake/release in astrocytes following treatment of cells with MBP charge isomers, Glutamate Assay Kit was used. The expression of EAAT-2 (excitatory amino acid transporters), peroxisome proliferator-activated receptor gamma (PPAR- γ), inhibitor of nuclear factor kappa B (IkB), and high mobility group protein B1 (HMGB1) in astrocytes were assayed by Western Blot analysis. Results: This study investigated the action of deiminated isomer (C8) on the cultured primary astrocytes and compared its effects with the effects of unmodified C1 isomers. The study found that C8 and C1 MBP differently act on the uptake and release of glutamate in astrocytes: nonmodified C1 MBP increases the uptake of glutamate and does not change the release, whereas C8 decreases the release of glutamate but does not alter the uptake. Nevertheless, both isomers increased the expression of PPAR-γ and EAAT2 in the same intensity. However, immunostaining and Western Blots of cell lysates showed a decrease of IkB and increased expression of HMGB1 after the treatment of astrocytes by C8. Moreover, in the presence of C8, astrocytes release more nitric oxide than unmodified C1 isomers. Conclusion: These data suggest that the deiminated isomer of MBP evokes an inflammatory response and enhances the ability of astrocytes to release proinflammatory mediators through activation of NF-kB after the breakdown of myelin sheets. Acknowledgment: This research was supported by the SRNSF Georgia RF17_534 grant.

Keywords: myelin basic protein, glutamate, deimination, astrocytes, inflammation

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Authors: Arpit Kumar Shrivastava, Subrat Kumar, Rajani Kanta Mohapatra, Priyadarshi Soumyaranjan Sahu

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Computational approaches to predict structure, function and other biological characteristics of proteins are becoming more common in comparison to the traditional methods in drug discovery. Cryptosporidiosis is a major zoonotic diarrheal disease particularly in children, which is caused primarily by Cryptosporidium hominis and Cryptosporidium parvum. Currently, there are no vaccines for cryptosporidiosis and recommended drugs are not effective. With the availability of complete genome sequence of C. hominis, new targets have been recognized for the development of effective and better drugs and/or vaccines. We identified a unique hypothetical epitopic protein in C. hominis genome through BLASTP analysis. A 3D model of the hypothetical protein was generated using I-Tasser server through threading methodology. The quality of the model was validated through Ramachandran plot by PROCHECK server. The functional annotation of the hypothetical protein through DALI server revealed structural similarity with human Transportin 3. Phylogenetic analysis for this hypothetical protein also showed C. hominis hypothetical protein (CUV04613) was the closely related to human transportin 3 protein. The 3D protein model is further subjected to virtual screening study with inhibitors from the Zinc Database by using Dock Blaster software. Docking study reported N-(3-chlorobenzyl) ethane-1,2-diamine as the best inhibitor in terms of docking score. Docking analysis elucidated that Leu 525, Ile 526, Glu 528, Glu 529 are critical residues for ligand–receptor interactions. The molecular dynamic simulation was done to access the reliability of the binding pose of inhibitor and protein complex using GROMACS software at 10ns time point. Trajectories were analyzed at each 2.5 ns time interval, among which, H-bond with LEU-525 and GLY- 530 are significantly present in MD trajectories. Furthermore, antigenic determinants of the protein were determined with the help of DNA Star software. Our study findings showed a great potential in order to provide insights in the development of new drug(s) or vaccine(s) for control as well as prevention of cryptosporidiosis among humans and animals.

Keywords: cryptosporidium hominis, hypothetical protein, molecular docking, molecular dynamics simulation

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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

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Authors: Michael P. Mannino, Gerald W. Hart

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The majority of glucose metabolism proceeds through glycolytic pathways such as glycolysis or pentose phosphate pathway, however, about 5% is shunted through the hexosamine biosynthetic pathway, producing uridine diphosphate N-acetyl glucosamine (UDP-GlcNAc). This precursor can then be incorporated into complex oligosaccharides decorating the cell surface or remain as an intracellular post-translational-modification (PTM) of serine/threonine residues (O-GlcNAcylation, OGN), which has been identified on over 4,000 cytosolic or nuclear proteins. Intracellular OGN has major implications on cellularprocesses, typically by modulating protein localization, protein-protein interactions, protein degradation, and gene expression. Additionally, OGN is known to have an extensive cross-talk with phosphorylation, be in a competitive or cooperative manner. Unlike other PTMs there are only two cycling enzymes that are capable of adding or removing the GlcNAc moiety, O-linked N-aceytl glucosamine Transferase (OGT) and O-linked N-acetyl glucoamidase (OGA), respectively. The activity of OGT has been shown to be sensitive to cellular UDP-GlcNAc levels, even changing substrate affinity. Owing to this and that the concentration of UDP-GlcNAc is related to the metabolisms of glucose, amino acid, fatty acid, and nucleotides, O-GlcNAc is often referred to as a nutrient sensing rheostat. Indeed OGN is known to regulate several signaling pathways as a result of nutrient levels, such as insulin signaling. Dysregulation of OGN is associated with several disease states such as cancer, diabetes, and neurodegeneration. Improvements in glycomics over the past 10-15 years has significantly increased the OGT substrate pool, suggesting O-GlcNAc’s involvement in a wide variety of signaling pathways. However, O-GlcNAc’s role at the receptor level has only been identified in a case-by-case basis of known pathways. Examining the OGN of the plasma membrane (PM) may better focus our understanding of O-GlcNAc-effected signaling pathways. In this current study, PM fractions were isolated from several cell types via ultracentrifugation, followed by purification and MS/MS analysis in several cell lines. This process was repeated with or without OGT/OGA inhibitors or with increased/decreased glucose levels in media to ascertain the importance of OGN. Various pathways are followed up on in more detailed studies employing methods to localize OGN at the PM specifically.

Keywords: GlcNAc, nutrient sensitive, post-translational-modification, receptor

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Authors: Richa Dhingra, Monika, Manav Malhotra, Tilak Raj Bhardwaj, Neelima Dhingra

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5-Alpha-reductases (5AR), a membrane bound, NADPH dependent enzyme and convert male hormone testosterone (T) into more potent androgen dihydrotestosterone (DHT). DHT is the required for the development and function of male sex organs, but its overproduction has been found to be associated with physiological conditions like Benign Prostatic Hyperplasia (BPH). Thus the inhibition of 5ARs could be a key target for the treatment of BPH. In present study, 2D and 3D Quantitative Structure Activity Relationship (QSAR) pharmacophore models have been generated for 5AR based on known inhibitory concentration (IC₅₀) values with extensive validations. The four featured 2D pharmacophore based PLS model correlated the topological interactions (–OH group connected with one single bond) (SsOHE-index); semi-empirical (Quadrupole2) and physicochemical descriptors (Mol. wt, Bromines Count, Chlorines Count) with 5AR inhibitory activity, and has the highest correlation coefficient (r² = 0.98, q² =0.84; F = 57.87, pred r² = 0.88). Internal and external validation was carried out using test and proposed set of compounds. The contribution plot of electrostatic field effects and steric interactions generated by 3D-QSAR showed interesting results in terms of internal and external predictability. The well validated 2D Partial Least Squares (PLS) and 3D k-nearest neighbour (kNN) models were used to search novel 5AR inhibitors with different chemical scaffold. To gain more insights into the molecular mechanism of action of these steroidal derivatives, molecular docking and in silico absorption, distribution, metabolism, and excretion (ADME) studies were also performed. Studies have revealed the hydrophobic and hydrogen bonding of the ligand with residues Alanine (ALA) 63A, Threonine (THR) 60A, and Arginine (ARG) 456A of 4AT0 protein at the hinge region. The results of QSAR, molecular docking, in silico ADME studies provide guideline and mechanistic scope for the identification of more potent 5-Alpha-reductase inhibitors (5ARI).

Keywords: 5α-reductase inhibitor, benign prostatic hyperplasia, ligands, molecular docking, QSAR

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Authors: Mateus L. O. Freitas, Flávia H. de M. Libório, Letycia L. Ricardo, Patrícia da C. Zonetti, Graciene de S. Bido

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Weeds compete with agricultural crops for resources such as light, water, and nutrients. This competition can cause significant damage to agricultural producers, and, currently, the use of agrochemicals is the most effective method for controlling these undesirable plants. Morning glory (Ipomoea grandifolia [Dammer] O'Donell) is an aggressive weed and significantly reduces agricultural productivity making harvesting difficult, especially mechanical harvesting. The biggest challenge in modern agriculture is to preserve high productivity reducing environmental damage and maintaining soil characteristics. No-till is a sustainable practice that can reduce the use of agrochemicals and environmental impacts due to the presence of plant residues in the soil, which release allelopathic compounds and reduce the incidence or alter the growth and development of crops and weeds. Sorghum (Sorghum bicolor [L.] Moench) is a forage with proven allelopathic activity, mainly for producing sorgholeone. In this context, this research aimed to evaluate the allelopathic action of sorghum fractions using hexane, dichloromethane, butanol, and ethyl acetate on the germination and initial growth of morning glory. The parameters analyzed were the percentage of germination, speed of germination, seedling length, and biomass weight (fresh and dry). The bioassays were performed in Petri dishes, kept in an incubation chamber for 7 days, at 25 °C, with a 12h photoperiod. The experimental design was completely randomized, with five replicates of each treatment. The data were evaluated by analysis of variance, and the averages between each treatment were compared using the Scott Knott test at a 5% significance level. The results indicated that the dichloromethane and ethyl acetate fractions showed bioherbicidal effects, promoting effective reductions on germination and initial growth of the morning glory. It was concluded that allelochemicals were probably extracted in these fractions. These secondary metabolites can reduce the use of agrochemicals and environmental impact, making agricultural production systems more sustainable.

Keywords: allelochemicals, secondary metabolism, sorgoleone, weeds

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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

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Authors: István Fekete, Imre Berki, Áron Béni, Katalin Juhos, Marianna Makádi, Zsolt Kotroczó

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The average annual precipitation significantly influences the moisture content of the soils and, through this, the decomposition of the organic substances in the soils, the leaching of nutrients from the soils, and the pH of the soils. Climate change, together with the lengthening of the vegetation period and the increasing CO₂ level, can increase the amount of biomass that is formed. Degradation processes, which accelerate as the temperature increases and slow down due to the drying climate, and the change in the degree of leaching can cancel out or strengthen each other's effects. In the course of our research, we looked for oak forests with climate-zonal soils where the geological, geographical and ecological background conditions are as similar as possible, apart from the different annual precipitation averages and the differences that can arise from them. We examined 5 dry and 5 humid Hungarian oak soils. Climate change affects the soils of drier and wetter forests differently. The aim of our research was to compare the content of carbon, nitrogen and some other nutrients, as well as the pH of the soils of humid and dry forests. Showing the effects of the drier climate on the tested soil parameters. In the case of the examined forest soils, we found a significant difference between the soils of dry and humid forests: in the case of the annual average precipitation values (p≥ 0.0001, for dry forest soils: 564±5.2 mm; for humid forest soils: 716±3.8 mm) for pH (p= 0.0004, for dry forest soils: 5.49±0.16; for wet forest soils: 5.36±0.21); for C content (p= 0.0054, for dry forest soils: 6.92%±0.59; for humid forest soils 3.09%±0.24), for N content (p= 0.0022, dry forest in the case of soils: 0.44%±0.047; in the case of humid forest soils: 0.23%±0.013), for the K content (p=0.0017, in the case of dry forest soils: 5684±732 (mg/kg); in the case of humid forest soils 2169±196 (mg/kg)), for the Ca content (p= 0.0096, for dry forest soils: 8207±2118 (mg/kg); for wet forest soils 957±320 (mg/kg)). No significant difference was found in the case of Mg. In a wetter environment, especially if the moisture content of the soil is also optimal for the decomposing organisms during the growing season, the decomposition of organic residues accelerates, and the processes of leaching from the soil are also intensified. The different intensity of the leaching processes is also well reflected in the quantitative differences of Ca and K, and in connection with these, it is also reflected in the difference in pH values. The differences in the C and N content can be explained by differences in the intensity of the decomposition processes. In addition to warming, drying is expected in a significant part of Hungary due to climate change. Thus, the comparison of the soils of dry and humid forests allows us to predict the subsequent changes in the case of the examined parameters.

Keywords: soil nutrients, precipitation difference, climate change, organic matter decomposition, leaching

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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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Authors: Zainab S. Ahmed, Mohammed S. Ali, Nadia A. Elshiekh, Sami Adam Ibrahim, Ghada M. El-Tayeb, Ahmed H. Elsadig, Rihab A. Omer, Sofia B. Mohamed

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Maple syrup urine (MSUD) is an autosomal recessive disease that causes a deficiency in the enzyme branched-chain alpha-keto acid (BCKA) dehydrogenase. The development of disease has been associated with SNPs in the DBT gene. Despite that, the computational analysis of SNPs in coding and noncoding and their functional impacts on protein level still remains unknown. Hence, in this study, we carried out a comprehensive in silico analysis of missense that was predicted to have a harmful influence on DBT structure and function. In this study, eight different in silico prediction algorithms; SIFT, PROVEAN, MutPred, SNP&GO, PhD-SNP, PANTHER, I-Mutant 2.0 and MUpo were used for screening nsSNPs in DBT including. Additionally, to understand the effect of mutations in the strength of the interactions that bind protein together the ELASPIC servers were used. Finally, the 3D structure of DBT was formed using Mutation3D and Chimera servers respectively. Our result showed that a total of 15 nsSNPs confirmed by 4 software (R301C, R376H, W84R, S268F, W84C, F276C, H452R, R178H, I355T, V191G, M444T, T174A, I200T, R113H, and R178C) were found damaging and can lead to a shift in DBT gene structure. Moreover, we found 7 nsSNPs located on the 2-oxoacid_dh catalytic domain, 5 nsSNPs on the E_3 binding domain and 3 nsSNPs on the Biotin Domain. So these nsSNPs may alter the putative structure of DBT’s domain. Furthermore, we detected all these nsSNPs are on the core residues of the protein and have the ability to change the stability of the protein. Additionally, we found W84R, S268F, and M444T have high significance, and they affected Leucine, Isoleucine, and Valine, which reduces or disrupt the function of BCKD complex, E2-subunit which the DBT gene encodes. In conclusion, based on our extensive in-silico analysis, we report 15 nsSNPs that have possible association with protein deteriorating and disease-causing abilities. These candidate SNPs can aid in future studies on Maple Syrup Urine Disease type II base in the genetic level.

Keywords: DBT gene, ELASPIC, in silico analysis, UCSF chimer

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Authors: Weixiang Liu, Yukun Qin, Song Liu, Pengcheng Li

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Plant diseases can cause the death of crops with great economic losses. Particularly, those diseases are usually caused by pathogenic fungi. Metal fungicides are a type of pesticide that has advantages of a low-cost, broad antimicrobial spectrum and strong sterilization effect. However, the frequent and wide application of traditional metal fungicides has caused serious problems such as environmental pollution, the outbreak of mites and phytotoxicity. Therefore, it is critically necessary to discover new organic metal fungicides alternatives that have a low metal content, low toxicity, and little influence on mites. Chitosan, the second most abundant natural polysaccharide next to cellulose, was proved to have broad-spectrum antifungal activity against a variety of fungi. However, the use of chitosan was limited due to its poor solubility and weaker antifungal activity compared with commercial fungicide. Therefore, in order to improve the water solubility and antifungal activity, many researchers grafted the active groups onto chitosan. The present work was to combine free metal ions with chitosan, to prepare more potent antifungal chitosan derivatives, thus, based on condensation reaction, chitosan derivative bearing amino pyridine group was prepared and subsequently followed by coordination with cupric ions, zinc ions and nickel ions to synthesize chitosan metal complexes. The calculations by density functional theory (DFT) show that the copper ions and nickel ions underwent dsp2 hybridization, the zinc ions underwent sp3 hybridization, and all of them are coordinated by the carbon atom in the p-π conjugate group and the oxygen atoms in the acetate ion. The antifungal properties of chitosan metal complexes against Phytophthora capsici (P. capsici), Gibberella zeae (G. zeae), Fusarium oxysporum (F. oxysporum) and Botrytis cinerea (B. cinerea) were also assayed. In addition, a plant toxicity experiment was carried out. The experiments indicated that the derivatives have significantly enhanced antifungal activity after metal ions complexation compared with the original chitosan. It was shown that 0.20 mg/mL of O-CSPX-Cu can 100% inhibit the growth of P. capsici and 0.20 mg/mL of O-CSPX-Ni can 87.5% inhibit the growth of B. cinerea. In general, their activities are better than the positive control oligosaccharides. The combination of the pyridine formyl groups seems to favor biological activity. Additionally, the ligand fashion was precisely analyzed, and the results revealed that the copper ions and nickel ions underwent dsp2 hybridization, the zinc ions underwent sp3 hybridization, and the carbon atoms of the p-π conjugate group and the oxygen atoms of acetate ion are involved in the coordination of metal ions. The phytotoxicity assay of O-CSPX-M was also conducted, unlike the traditional metal fungicides, the metal complexes were not significantly toxic to the leaves of wheat. O-CSPX-Zn can even increase chlorophyll content in wheat leaves at 0.40 mg/mL. This is mainly because chitosan itself promotes plant growth and counteracts the phytotoxicity of metal ions. The chitosan derivative described here may lend themselves to future applicative studies in crop protection.

Keywords: coordination, chitosan, metal complex, antifungal properties

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Authors: Francielo Vendruscolo

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During industrial processing of raw materials of animal and vegetable origin, large amounts of solid, liquid and gaseous wastes are generated. Solid residues are usually materials rich in carbohydrates, protein, fiber and minerals. Brewer’s spent grain (BSG) is the main waste generated in the brewing industry, representing 85% of the waste generated in this industry. It is estimated that world’s BSG generation is approximately 38.6 x 106 t per year and represents 20-30% (w/w) of the initial mass of added malt, resulting in low commercial value by-product, however, does not have economic value, but it must be removed from the brewery, as its spontaneous fermentation can attract insects and rodents. For every 100 grams in dry basis, BSG has approximately 68 g total fiber, being divided into 3.5 g of soluble fiber and 64.3 g of insoluble fiber (cellulose, hemicellulose and lignin). In addition to dietary fibers, depending on the efficiency of the grinding process and mashing, BSG may also have starch, reducing sugars, lipids, phenolics and antioxidants, emphasizing that its composition will depend on the barley variety and cultivation conditions, malting and technology involved in the production of beer. BSG demands space for storage, but studies have proposed alternatives such as the use of drying, extrusion, pressing with superheated steam, and grinding to facilitate storage. Other important characteristics that enhance its applicability in bioremediation, effluent treatment and biotechnology, is the surface area (SBET) of 1.748 m2 g-1, total pore volume of 0.0053 cm3 g-1 and mean pore diameter of 121.784 Å, characterized as a macroporous and possess fewer adsorption properties but have great ability to trap suspended solids for separation from liquid solutions. It has low economic value; however, it has enormous potential for technological applications that can improve or add value to this agro-industrial waste. Due to its composition, this material has been used in several industrial applications such as in the production of food ingredients, fiber enrichment by its addition in foods such as breads and cookies in bioremediation processes, substrate for microorganism and production of biomolecules, bioenergy generation, and civil construction, among others. Therefore, the use of this waste or by-product becomes essential and aimed at reducing the amount of organic waste in different industrial processes, especially in breweries.

Keywords: brewer’s spent malt, agro-industrial residue, lignocellulosic material, waste generation

Procedia PDF Downloads 208

Authors: Marzouk Benali, Jawad Jeaidi, Behrang Mansoornejad, Olumoye Ajao, Banafsheh Gilani, Nima Ghavidel Mehr

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Canadian forest industry is seeking to identify and implement transformational strategies for enhanced financial performance through the emerging bioeconomy or more specifically through the concept of the biorefinery. For example, processing forest residues or surplus of biomass available on the mill sites for the production of biofuels, biochemicals and/or biomaterials is one of the attractive strategies along with traditional wood and paper products and cogenerated energy. There are many possible process-product biorefinery pathways, each associated with specific product portfolios with different levels of risk. Thus, it is not obvious which unique strategy forest industry should select and implement. Therefore, there is a need for analytical and design tools that enable evaluating biorefinery strategies based on a set of criteria considering a perspective of sustainability over the short and long terms, while selecting the existing core products as well as selecting the new product portfolio. In addition, it is critical to assess the manufacturing flexibility to internalize the risk from market price volatility of each targeted bio-based product in the product portfolio, prior to invest heavily in any biorefinery strategy. The proposed paper will focus on introducing a systematic methodology for designing integrated biorefineries using process systems engineering tools as well as a multi-criteria decision making framework to put forward the most effective biorefinery strategies that fulfill the needs of the forest industry. Topics to be covered will include market analysis, techno-economic assessment, cost accounting, energy integration analysis, life cycle assessment and supply chain analysis. This will be followed by describing the vision as well as the key features and functionalities of the I-BIOREF software platform, developed by CanmetENERGY of Natural Resources Canada. Two industrial case studies will be presented to support the robustness and flexibility of I-BIOREF software platform: i) An integrated Canadian Kraft pulp mill with lignin recovery process (namely, LignoBoost™); ii) A standalone biorefinery based on ethanol-organosolv process.

Keywords: biorefinery strategies, bioproducts, co-production, multi-criteria decision making, tool

Procedia PDF Downloads 232

Authors: Laura M. Hovsepyan, Gayane S. Ghazaryan, Hasmik V. Zanginyan

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Hypertension (HD) is the most common cardiovascular pathology that causes disability and mortality in the working population. Most often, heart failure (HF), which is based on myocardial remodeling, leads to death in hypertension. Recently, endothelial dysfunction (EDF) or a violation of the functional state of the vascular endothelium has been assigned a significant role in the structural changes in the myocardium and the occurrence of heart failure in patients with hypertension. It has now been established that tissues affected by inflammation form increased amounts of superoxide radical and NO, which play a significant role in the development and pathogenesis of various pathologies. They mediate inflammation, modify proteins and damage nucleic acids. The aim of this work was to study the processes of oxidative modification of proteins (OMP) and the production of nitric oxide in hypertension. In the experimental work, the blood of 30 donors and 33 patients with hypertension was used. For the quantitative determination of OMP products, the based on the reaction of the interaction of oxidized amino acid residues of proteins and 2,4-dinitrophenylhydrazine (DNPH) with the formation of 2,4-dinitrophenylhydrazones, the amount of which was determined spectrophotometrically. The optical density of the formed carbonyl derivatives of dinitrophenylhydrazones was recorded at different wavelengths: 356 nm - aliphatic ketone dinitrophenylhydrazones (KDNPH) of neutral character; 370 nm - aliphatic aldehyde dinirophenylhydrazones (ADNPH) of neutral character; 430 nm - aliphatic KDNFG of the main character; 530 nm - basic aliphatic ADNPH. Nitric oxide was determined by photometry using Grace's solution. Adsorption was measured on a Thermo Scientific Evolution 201 SF at a wavelength of 546 nm. Thus, the results of the studies showed that in patients with arterial hypertension, an increased level of nitric oxide in the blood serum is observed, and there is also a tendency to an increase in the intensity of oxidative modification of proteins at a wavelength of 270 nm and 363 nm, which indicates a statistically significant increase in aliphatic aldehyde and ketone dinitrophenylhydrazones. The increase in the intensity of oxidative modification of blood plasma proteins in the studied patients, revealed by us, actually reflects the general direction of free radical processes and, in particular, the oxidation of proteins throughout the body. A decrease in the activity of the antioxidant system also leads to a violation of protein metabolism. The most important consequence of the oxidative modification of proteins is the inactivation of enzymes.

Keywords: hypertension (HD), oxidative modification of proteins (OMP), nitric oxide (NO), oxidative stress

Procedia PDF Downloads 108

Authors: Olfat A. Mohamed

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Rhamnolipids biosurfactants have distinct properties given them importance in many industrial applications, especially their great new future applications in cosmetic and pharmaceutical industries. These applications have encouraged the search for diverse and renewable resources to control the cost of production. The experimental results were then applied to find a suitable mathematical model for obtaining the design criteria of the batch bioreactor. This research aims to produce Rhamnolipids from different oily wastewater sources such as petroleum crude oil (PO) and vegetable oil (VO) by using Pseudomonas aeruginosa ATCC 9027. Different concentrations of the PO and the VO are added to the media broth separately are in arrangement (0.5 1, 1.5, 2, 2.5 % v/v) and (2, 4, 6, 8 and 10%v/v). The effect of the initial concentration of oil residues and the addition of glycerol and palmitic acid was investigated as an inducer in the production of rhamnolipid and the surface tension of the broth. It was found that 2% of the waste (PO) and 6% of the waste (VO) was the best initial substrate concentration for the production of rhamnolipids (2.71, 5.01 g rhamnolipid/l) as arrangement. Addition of glycerol (10-20% v glycerol/v PO) to the 2% PO fermentation broth led to increase the rhamnolipid production (about 1.8-2 times fold). However, the addition of palmitic acid (5 and 10 g/l) to fermentation broth contained 6% VO rarely enhanced the production rate. The experimental data for 2% initially (PO) was used to estimate the various kinetic parameters. The following results were obtained, maximum rate or velocity of reaction (Vmax) = 0.06417 g/l.hr), yield of cell weight per unit weight of substrate utilized (Yx/s = 0.324 g Cx/g Cs) maximum specific growth rate (μmax = 0.05791 hr⁻¹), yield of rhamnolipid weight per unit weight of substrate utilized (Yp/s)=0.2571gCp/g Cs), maintenance coefficient (Ms =0.002419), Michaelis-Menten constant, (Km=6.1237 gmol/l), endogenous decay coefficient (Kd=0.002375 hr⁻¹). Predictive parameters and advanced mathematical models were applied to evaluate the time of the batch bioreactor. The results were as follows: 123.37, 129 and 139.3 hours in respect of microbial biomass, substrate and product concentration, respectively compared with experimental batch time of 120 hours in all cases. The expected mathematical models are compatible with the laboratory results and can, therefore, be considered as tools for expressing the actual system.

Keywords: batch bioreactor design, glycerol, kinetic parameters, petroleum crude oil, Pseudomonas aeruginosa, rhamnolipids biosurfactants, vegetable oil

Procedia PDF Downloads 131

Authors: Roya Bakhtiar, Alireza Abdolmohammadi, Hadi Hajarian, Zahra Nikousefat, Davood, Kalantar-Neyestanaki

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The objective of the present study was to determine the polymorphism in the leptin (332G>A) and its association with biometric traits in Sanjabi sheep. For this purpose, blood samples from 96 rams were taken, and tail length, width tail, circumference tail, body length, body width, and height were simultaneously recorded. PCR was performed using specific primer to amplify 463 bp fragment including exon 3 of leptin gene, and PCR products were digested by Cail restriction enzymes. The 332G>A (at 332th nucleotide of exon 3 leptin gene) that caused an amino acid change from Arg to Gln was detected by Cail (CAGNNNCTG) endonuclease, as the endonuclease cannot cut this region if G nucleotide is located in this position. Three genotypes including GG (463), GA (463, 360and 103 bp) and GG (360 bp and 103 bp) were identified after digestion by enzyme. The estimated frequencies of three genotypes including GG, GA, and AA for 332G>A locus were 0.68, 0.29 and 0.03 and those were 0.18 and 0.82 for A and G alleles, respectively. In the current study, chi-square test indicated that 332G>A positions did not deviate from the Hardy–Weinberg (HW) equilibrium. The most important reason to show HW equation was that samples used in this study belong to three large local herds with a traditional breeding system having random mating and without selection. Shannon index amount was calculated which represent an average genetic variation in Sanjabi rams. Also, heterozygosity estimated by Nei index indicated that genetic diversity of mutation in the leptin gene is moderate. Leptin gene polymorphism in the 332G>A had significant effect on body length (P<0.05) trait, and individuals with GA genotype had significantly the higher body length compared to other individuals. Although animals with GA genotype had higher body width, this difference was not statistically significant (P>0.05). This non-synonymous SNP resulted in different amino acid changes at codon positions111(R/Q). As leptin activity is localized, at least in part, in domains between amino acid residues 106-1406, it is speculated that the detected SNP at position 332 may affect the activity of leptin and may lead to different biological functions. Based to our results, due to significant effect of leptin gene polymorphism on body size traits, this gene may be used a candidate gene for improving these traits.

Keywords: body size, Leptin gene, PCR-RFLP, Sanjabi sheep

Procedia PDF Downloads 341

Authors: Farhan J. M. Al-Behadili, Vineeta Bilgi, Miyuki Taniguchi, Junxi Li, Wei Xu

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Mediterranean fruit fly (Ceratitis capitata) is one of the most destructive pests of fruits and vegetables. Medfly originated from Africa and spread in many countries, and is currently an endemic pest in Western Australia. Medfly has been recorded from over 300 plant species including fruits, vegetables, nuts and its main hosts include blueberries, citrus, stone fruit, pome fruits, peppers, tomatoes, and figs. Global trade of fruits and other farm fresh products are suffering from the damages of this pest, which prompted towards the need to develop more effective ways to control these pests. The available quarantine treatment technologies mainly include chemical treatment (e.g., fumigation) and non-chemical treatments (e.g., cold, heat and irradiation). In recent years, with the loss of several chemicals, it has become even more important to rely on non-chemical postharvest control technologies (i.e., heat, cold and irradiation) to control fruit flies. Cold treatment is one of the most potential trends of focus in postharvest treatment because it is free of chemical residues, mitigates or kills the pest population, increases the strength of the fruits, and prolongs storage time. It can also be applied to fruits after packing and ‘in transit’ during lengthy transport by sea during their exports. However, limited systematic study on cold treatment of Medfly stages in artificial diets was reported, which is critical to provide a scientific basis to compare with previous research in plant products and design an effective cold treatment suitable for exported plant products. The overall purpose of this study was to evaluate and understand Medfly responses to cold treatments. Medfly stages were tested. The long-term goal was to optimize current postharvest treatments and develop more environmentally-friendly, cost-effective, and efficient treatments for controlling Medfly. Cold treatment with different exposure times is studied to evaluate cold eradication treatment of Mediterranean fruit fly (Ceratitis capitata), that reared on carrot diet. Mortality is important aspect was studied in this study. On the other hand, study effects of exposure time on mortality means of medfly stages.

Keywords: cold treatment, fruit fly, Ceratitis capitata, carrot diet, temperature effects

Procedia PDF Downloads 224

Authors: Shah Rucksana Akhter Urme

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Agriculture is the main source of food for human consumption and feeding the world huge population, the pressure of food supply is increasing day by day. Undoubtedly, quality strain, improved plantation, farming technology, synthetic fertilizer, readily available irrigation, insecticides and harvesting technology are the main factors those to meet up the huge demand of food consumption all over the world. However, depended on this limited resources and excess amount of consuming lands, water, fertilizers leads to the end of the resources and severe climate effects has been left for our future generation. Agriculture is the most responsible to global warming, emitting more greenhouse gases than all other vehicles largely from nitrous oxide released by from fertilized fields, and carbon dioxide from the cutting of rain forests to grow crops . Farming is the thirstiest user of our precious water supplies and a major polluter, as runoff from fertilizers disrupts fragile lakes, rivers, and coastal ecosystems across the globe which accelerates the loss of biodiversity, crucial habitat and a major driver of wildlife extinction. It is needless to say that we have to more concern on how we can save the nutrients of the soil, storage of the water and avoid excessive depends on synthetic fertilizer and insecticides. In this case, eco- friendly cultivation could be a potential alternative solution to minimize effects of agriculture in our environment. The objective of this review paper is about organic cultivation following in particular biotechnological process focused on bio-fertilizer and bio-pesticides. Intense practice of chemical pesticides, insecticides has severe effect on both in human life and biodiversity. This cultivation process introduces farmer an alternative way which is nonhazardous, cost effective and ecofriendly. Organic fertilizer such as tea residue, ashes might be the best alternative to synthetic fertilizer those play important role in increasing soil nutrient and fertility. Ashes contain different essential and non-essential mineral contents that are required for plant growth. Organic pesticide such as neem spray is beneficial for crop as it is toxic for pest and insects. Recycled and composted crop wastes and animal manures, crop rotation, green manures and legumes etc. are suitable for soil fertility which is free from hazardous chemicals practice. Finally water hyacinth and algae are potential source of nutrients even alternative to soil for cultivation along with storage of water for continuous supply. Inorganic practice of agriculture, consuming fruits and vegetables becomes a threat for both human life and eco-system and synthetic fertilizer and pesticides are responsible for it. Farmers that practice eco-friendly farming have to implement steps to protect the environment, particularly by severely limiting the use of pesticides and avoiding the use of synthetic chemical fertilizers, which are necessary for organic systems to experience reduced environmental harm and health risk.

Keywords: organic farming, biopesticides, organic nutrients, water storage, global warming

Procedia PDF Downloads 60

Authors: Choupani Andisheh, Temucin Elif Sevval, Bediz Bekir

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Due to their various advantages compared to many other drug delivery systems such as hypodermic injections and oral medications, microneedle arrays (MNAs) are a promising drug delivery system. To achieve enhanced performance of the MN, it is crucial to develop numerical models, optimization methods, and simulations. Accordingly, in this work, the optimized design of dissolvable MNAs, as well as their manufacturing, is investigated. For this purpose, a mechanical model of a single MN, having the geometry of an obelisk, is developed using commercial finite element software. The model considers the condition in which the MN is under pressure at the tip caused by the reaction force when penetrating the skin. Then, a multi-objective optimization based on non-dominated sorting genetic algorithm II (NSGA-II) is performed to obtain geometrical properties such as needle width, tip (apex) angle, and base fillet radius. The objective of the optimization study is to reach a painless and effortless penetration into the skin along with minimizing its mechanical failures caused by the maximum stress occurring throughout the structure. Based on the obtained optimal design parameters, master (male) molds are then fabricated from PMMA using a mechanical micromachining process. This fabrication method is selected mainly due to the geometry capability, production speed, production cost, and the variety of materials that can be used. Then to remove any chip residues, the master molds are cleaned using ultrasonic cleaning. These fabricated master molds can then be used repeatedly to fabricate Polydimethylsiloxane (PDMS) production (female) molds through a micro-molding approach. Finally, Polyvinylpyrrolidone (PVP) as a dissolvable polymer is cast into the production molds under vacuum to produce the dissolvable MNAs. This fabrication methodology can also be used to fabricate MNAs that include bioactive cargo. To characterize and demonstrate the performance of the fabricated needles, (i) scanning electron microscope images are taken to show the accuracy of the fabricated geometries, and (ii) in-vitro piercing tests are performed on artificial skin. It is shown that optimized MN geometries can be precisely fabricated using the presented fabrication methodology and the fabricated MNAs effectively pierce the skin without failure.

Keywords: microneedle, microneedle array fabrication, micro-manufacturing structural optimization, finite element analysis

Procedia PDF Downloads 113

Authors: Mónica P. Cala, Juan S. Carreño, Roland J.W. Meesters

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Recently, collection of biological fluids on special filter papers has become a popular micro-sampling technique. Especially, the dried blood spot (DBS) micro-sampling technique has gained much attention and is momently applied in various life sciences reserach areas. As a result of this popularity, DBS are not only intensively competing with the venous blood sampling method but are at this moment widely applied in numerous bioanalytical assays. In particular, in the screening of inherited metabolic diseases, pharmacokinetic modeling and in therapeutic drug monitoring. Recently, microsampling techniques were also introduced in “omics” areas, whereunder metabolomics. For a metabolic profiling study we applied micro-sampling of biological fluids (blood and plasma) from healthy controls and from women with breast cancer. From blood samples, dried blood and plasma samples were prepared by spotting 8uL sample onto pre-cutted 5-mm paper disks followed by drying of the disks for 100 minutes. Dried disks were then extracted by 100 uL of methanol. From liquid blood and plasma samples 40 uL were deproteinized with methanol followed by centrifugation and collection of supernatants. Supernatants and extracts were evaporated until dryness by nitrogen gas and residues derivated by O-methyxyamine and MSTFA. As internal standard C17:0-methylester in heptane (10 ppm) was used. Deconvolution and alignment of and full scan (m/z 50-500) MS data were done by AMDIS and SpectConnect (http://spectconnect.mit.edu) software, respectively. Statistical Data analysis was done by Principal Component Analysis (PCA) using R software. The results obtained from our preliminary study indicate that the use of dried blood/plasma on paper disks could be a powerful new tool in metabolic profiling. Many of the metabolites observed in plasma (liquid/dried) were also positively identified in whole blood samples (liquid/dried). Whole blood could be a potential substitute matrix for plasma in Metabolomic profiling studies as well also micro-sampling techniques for the collection of samples in clinical studies. It was concluded that the separation of the different sample methodologies (liquid vs. dried) as observed by PCA was due to different sample treatment protocols applied. More experiments need to be done to confirm obtained observations as well also a more rigorous validation .of these micro-sampling techniques is needed. The novelty of our approach can be found in the application of different biological fluid micro-sampling techniques for metabolic profiling.

Keywords: biofluids, breast cancer, metabolic profiling, micro-sampling

Procedia PDF Downloads 411

Authors: Walter W. Loo

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China declared the “zero discharge rules which leave no toxics into our living environment and deliver blue sky, green land and clean water to many generations to come”. The achievement of ZWD will provide conservation of water, soil and energy and provide drastic increase in Gross Domestic Products (GDP). Our society’s engine needs a major tune up; it is sputtering. ZWD is achieved in world’s space stations – no toxic air emission and the water is totally recycled and solid wastes all come back to earth. This is all done with solar power. These are all achieved under extreme temperature, pressure and zero gravity in space. ZWD can be achieved on earth under much less fluctuations in temperature, pressure and normal gravity environment. ZWD systems are not expensive and will have multiple beneficial returns on investment which are both financially and environmentally acceptable. The paper will include successful case histories since the mid-1970s. ZWD discharge can be applied to the following types of projects: nuclear and coal fire power plants with a closed loop system that will eliminate thermal water discharge; residential communities with wastewater treatment sump and recycle the water use as a secondary water supply; waste water treatment Plants with complete water recycling including water distillation to produce distilled water by very economical 24-hours solar power plant. Landfill remediation is based on neutralization of landfilled gas odor and preventing anaerobic leachate formation. It is an aerobic condition which will render landfill gas emission explosion proof. Desert development is the development of recovering soil moisture from soil and completing a closed loop water cycle by solar energy within and underneath an enclosed greenhouse. Salt-alkali land development can be achieved by solar distillation of salty shallow water into distilled water. The distilled water can be used for soil washing and irrigation and complete a closed loop water cycle with energy and water conservation. Heavy metals remediation can be achieved by precipitation of dissolved toxic metals below the plant or vegetation root zone by solar electricity without pumping and treating. Soil and groundwater remediation - abandoned refineries, chemical and pesticide factories can be remediated by in-situ electrobiochemical and bioventing treatment method without pumping or excavation. Toxic organic chemicals are oxidized into carbon dioxide and heavy metals precipitated below plant and vegetation root zone. New water sources: low temperature distilled water can be recycled for repeated use within a greenhouse environment by solar distillation; nano bubble water can be made from the distilled water with nano bubbles of oxygen, nitrogen and carbon dioxide from air (fertilizer water) and also eliminate the use of pesticides because the nano oxygen will break the insect growth chain in the larvae state. Three dimensional high yield greenhouses can be constructed by complete water recycling using the vadose zone soil as a filter with no farming wastewater discharge.

Keywords: greenhouses, no discharge, remediation of soil and water, wastewater

Procedia PDF Downloads 344

Authors: Elif Tugce Aksun Tumerkan, Chris Lowe, Hannah Krupa

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Fluorescent proteins are self-sufficient in forming chromophores with a visible wavelength from 3 amino acids sequence within their own polypeptide structure. This chromophore – a molecule that absorbs a photon of light and exhibits an energy transition equal to the energy of the absorbed photon. Fluorescent proteins (FPs) consisted of a chain of 238 amino acid residues and composed of 11 beta strands shaped in a cylinder surrounding an alpha helix structure. A better understanding of the system of the chromospheres and the increasing advance in protein engineering in recent years, the properties of FPs offers the potential for new applications. They have used sensors and probes in molecular biology and cell-based research that giving a chance to observe these FPs tagged cell localization, structural variation and movement. For clarifying functional uses of fluorescent proteins, electrophoretic properties of these proteins are one of the most important parameters. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis is used for determining electrophoretic properties commonly. While there are many techniques are used for determining the functionality of protein-based research, SDS-PAGE analysis can only provide a molecular level assessment of the proteolytic fragments. Before SDS-PAGE analysis, fluorescent proteins need to successfully purified. Due to directly purification of the target, FPs is difficult from the animal, gene expression is commonly used which must be done by transformation with the plasmid. Furthermore, used gel within electrophoresis and staining agents properties have a key role. In this review, the different factors that have the impact on the electrophoretic properties of fluorescent proteins explored. Fluorescent protein separation and purification are the essential steps before electrophoresis that should be done very carefully. For protein purification, gene expression process and following steps have a significant function. For successful gene expression, the properties of selected bacteria for expression, used plasmid are essential. Each bacteria has own characteristics which are very sensitive to gene expression, also used procedure is the important factor for fluorescent protein expression. Another important factors are gel formula and used staining agents. Gel formula has an effect on the specific proteins mobilization and staining with correct agents is a key step for visualization of electrophoretic bands of protein. Visuality of proteins can be changed depending on staining reagents. Apparently, this review has emphasized that gene expression and purification have a stronger effect than electrophoresis protocol and staining agents.

Keywords: cell biology, gene expression, staining agents, SDS-page

Procedia PDF Downloads 194

Authors: Andrea M. Rivas-Castillo, Marlenne Gómez-Ramirez, Isela Rodríguez-Pozos, Norma G. Rojas-Avelizapa

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Spent catalysts are considered as hazardous residues of major concern, mainly due to the simultaneous presence of several metals in elevated concentrations. Although hydrometallurgical, pyrometallurgical and chelating agent methods are available to remove and recover some metals contained in spent catalysts; these procedures generate potentially hazardous wastes and the emission of harmful gases. Thus, biotechnological treatments are currently gaining importance to avoid the negative impacts of chemical technologies. To this end, diverse microorganisms have been used to assess the removal of metals from spent catalysts, comprising bacteria, archaea and fungi, whose resistance and metal uptake capabilities differ depending on the microorganism tested. Acidophilic sulfur oxidizing bacteria have been used to investigate the biotreatment and extraction of valuable metals from spent catalysts, namely Acidithiobacillus thiooxidans and Acidithiobacillus ferroxidans, as they present the ability to produce leaching agents such as sulfuric acid and sulfur oxidation intermediates. In the present work, the ability of A. thiooxidans DSM 26636 for the bioleaching of metals contained in five different spent catalysts was assessed by growing the culture in modified Starkey mineral medium (with elemental sulfur at 1%, w/v), and 1% (w/v) pulp density of each residue for up to 21 days at 30 °C and 150 rpm. Sulfur-oxidizing activity was periodically evaluated by determining sulfate concentration in the supernatants according to the NMX-k-436-1977 method. The production of sulfuric acid was assessed in the supernatants as well, by a titration procedure using NaOH 0.5 M with bromothymol blue as acid-base indicator, and by measuring pH using a digital potentiometer. On the other hand, Inductively Coupled Plasma - Optical Emission Spectrometry was used to analyze metal removal from the five different spent catalysts by A. thiooxidans DSM 26636. Results obtained show that, as could be expected, sulfuric acid production is directly related to the diminish of pH, and also to highest metal removal efficiencies. It was observed that Al and Fe are recurrently removed from refinery spent catalysts regardless of their origin and previous usage, although these removals may vary from 9.5 ± 2.2 to 439 ± 3.9 mg/kg for Al, and from 7.13 ± 0.31 to 368.4 ± 47.8 mg/kg for Fe, depending on the spent catalyst proven. Besides, bioleaching of metals like Mg, Ni, and Si was also obtained from automotive spent catalysts, which removals were of up to 66 ± 2.2, 6.2±0.07, and 100±2.4, respectively. Hence, the data presented here exhibit the potential of A. thiooxidans DSM 26636 for the simultaneous bioleaching of metals contained in spent catalysts from diverse provenance.

Keywords: bioleaching, metal removal, spent catalysts, Acidithiobacillus thiooxidans

Procedia PDF Downloads 140

Authors: A. B. Moldes, X. Vecino, L. Rodriguez-López, J. M. Dominguez, J. M. Cruz

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The concept of the circular economy is focus on the importance of providing goods and services sustainably. Thus, in a future it will be necessary to respond to the environmental contamination and to the use of renewables substrates by moving to a more restorative economic system that drives towards the utilization and revalorization of residues to obtain valuable products. During its evolution our industrial economy has hardly moved through one major characteristic, established in the early days of industrialization, based on a linear model of resource consumption. However, this industrial consumption system will not be maintained during long time. On the other hand, there are many industries, like the corn milling industry, that although does not consume high amount of non renewable substrates, they produce valuable streams that treated accurately, they could provide additional, economical and environmental, benefits by the extraction of interesting commercial renewable products, that can replace some of the substances obtained by chemical synthesis, using non renewable substrates. From this point of view, the use of streams from corn milling industry to obtain surface-active compounds will decrease the utilization of non-renewables sources for obtaining this kind of compounds, contributing to a circular and global economy. However, the success of the circular economy depends on the interest of the industrial sectors in the revalorization of their streams by developing relevant and new business models. Thus, it is necessary to invest in the research of new alternatives that reduce the consumption of non-renewable substrates. In this study is proposed the utilization of a corn milling industry stream to obtain an extract with surfactant capacity. Once the biosurfactant is extracted, the corn milling stream can be commercialized as nutritional media in biotechnological process or as animal feed supplement. Usually this stream is combined with other ingredients obtaining a product namely corn gluten feed or may be sold separately as a liquid protein source for beef and dairy feeding, or as a nutritional pellet binder. Following the productive scheme proposed in this work, the corn milling industry will obtain a biosurfactant extract that could be incorporated in its productive process replacing those chemical detergents, used in some point of its productive chain, or it could be commercialized as a new product of the corn manufacture. The biosurfactants obtained from corn milling industry could replace the chemical surfactants in many formulations, and uses, and it supposes an example of the potential that many industrial streams could offer for obtaining valuable products when they are manage properly.

Keywords: biosurfactantes, circular economy, corn, sustainability

Procedia PDF Downloads 261

Authors: Vasiliki Kolake, Christos Kavalaris, Sofia Megoudi, Maria Maxouri, Panagiotis A. Karas, Aris Kyparissis, Efi Levizou

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Conservation agriculture (CA), is a production system approach that can tackle the challenges of climate change mainly through facilitating carbon storage into the soil and increasing crop resilience. This is extremely important for the vulnerable Mediterranean agroecosystems, which already face adverse environmental conditions. The agronomic practices used in CA, i.e. permanent soil cover and no-tillage, result in reduced soil erosion and increased soil organic matter, preservation of water and improvement of quality and fertility of the soil in the long-term. Thus the functional characteristics and processes of the soil are considerably affected by the implementation of CA. The aim of the present work was to assess the effects of CA on soil nitrification potential and mycorrhizal colonization about the above-ground production in a wheat field. Two adjacent but independent field sites of 1.5ha each were used (Thessaly plain, Central Greece), comprising the no-till and conventional tillage treatments. The no-tillage site was covered by residues of the previous crop (cotton). Potential nitrification and the nitrate and ammonium content of the soil were measured at two different soil depths (3 and 15cm) at 20-days intervals throughout the growth period. Additionally, the leaf area index (LAI) was monitored at the same time-course. The mycorrhizal colonization was measured at the commencement and end of the experiment. At the final harvest, total yield and plant biomass were also recorded. The results indicate that wheat yield was considerably favored by CA practices, exhibiting a 42% increase compared to the conventional tillage treatment. The superior performance of the CA crop was also depicted in the above-ground plant biomass, where a 26% increase was recorded. LAI, which is considered a reliable growth index, did not show statistically significant differences between treatments throughout the growth period. On the contrary, significant differences were recorded in endomycorrhizal colonization one day before the final harvest, with CA plants exhibiting 20% colonization, while the conventional tillage plants hardly reached 1%. The on-going analyses of potential nitrification measurements, as well as nitrate and ammonium determination, will shed light on the effects of CA on key processes in the soil. These results will integrate the assessment of CA impact on certain below and above-ground processes during wheat cultivation under the Mediterranean climate.

Keywords: conservation agriculture, LAI, mycorrhizal colonization, potential nitrification, wheat, yield

Procedia PDF Downloads 130

Authors: Z. C. Dlamini, R. L. S. Langa, A. I. Okoh, O. A. Aiyegoro

Abstract:

The indiscriminate usage of antibiotics in swine production have consequential outcomes; such as development of bacterial resistance to prophylactic antibiotics and possibility of antibiotic residues in animal products. The use of probiotics appears to be the most effective procedure with positive metabolic nutritional implications. The aim of this study was to investigate the efficacy of probiotic bacteria (Lactobacillus reuteri ZJ625, Lactobacillus reuteri VB4, Lactobacillus salivarius ZJ614 and Streptococcus salivarius NBRC13956) administered as direct-fed microorganisms in weaned piglets. 45 weaned piglets blocked by weight were dived into 5 treatments groups: diet with antibiotic, diet with no-antibiotic and no probiotic, and diet with probiotic and diet with combination of probiotics. Piglets performance was monitored during the trials. Faecal and Ileum samples were collected for microbial count analysis. Blood samples were collected from pigs at the end of the trial, for analysis of haematological, biochemical and IgG stimulation. The data was analysed by Split-Plot ANOVA using SAS statistically software (SAS 9.3) (2003). The difference was observed between treatments for daily weight and feed conversion ratio. No difference was observed in analysis of faecal samples in regards with bacterial counts, difference was observed in ileums samples with enteric bacteria colony forming unit being lower in P2 treatment group as compared with lactic acid and total bacteria. With exception of globulin and albumin, biochemistry blood parameters were not affected, likewise for haematology, only basophils and segmented neutrophils were differed by having higher concentration in NC treatment group as compared with other treatment groups. Moreover, in IgG stimulation analysis, difference was also observed, with P2 treatment group having high concentration of IgG in P2 treatment group as compared to other groups. The results of this study suggest that probiotics have a beneficial effect on growth performances, blood parameters and IgG stimulation of pigs, most effective when they are administered in synergy form. This means that it is most likely that these probiotics will offer a significant benefit in pig farming by reducing risk of morbidity and mortality and produce quality meat that is more affordable to poorer communities, and thereby enhance South African pig industry’s economy. In addition, these results indicate that there is still more research need to be done on probiotics in regards with, i.e. dosage, shelf life and mechanism of action.

Keywords: antibiotics, biochemistry, haematology, IgG-stimulation, microbial count, probiotics

Procedia PDF Downloads 301

Authors: Tais Basaco, Stefanie Pektor, Josue A. Moreno, Matthias Miederer, Andreas Türler

Abstract:

Radiopharmaceuticals based in monoclonal anti-body (mAb) via chemical linkers have become a potential tool in nuclear medicine because of their specificity and the large variability and availability of therapeutic radiometals. It is important to identify the conjugation sites and number of attached chelator to mAb to obtain radioimmunoconjugates with required immunoreactivity and radiostability. Girentuximab antibody (G250) is a potential candidate for radioimmunotherapy of clear cell carcinomas (RCCs) because it is reactive with CAIX antigen, a transmembrane glycoprotein overexpressed on the cell surface of most ( > 90%) (RCCs). G250 was conjugated with the bifunctional chelating agent DOTA (1,4,7,10-Tetraazacyclododecane-N,N’,N’’,N’’’-tetraacetic acid) via a benzyl-thiocyano group as a linker (p-SCN-Bn-DOTA). DOTA-G250 conjugates were analyzed by size exclusion chromatography (SE-HPLC) and by electrophoresis (SDS-PAGE). The potential site-specific conjugation was identified by liquid chromatography–mass spectrometry (LC/MS-MS) and the number of linkers per molecule of mAb was calculated using the molecular weight (MW) measured by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The average number obtained in the conjugates in non-reduced conditions was between 8-10 molecules of DOTA per molecule of mAb. The average number obtained in the conjugates in reduced conditions was between 1-2 and 3-4 molecules of DOTA per molecule of mAb in the light chain (LC) and heavy chain (HC) respectively. Potential DOTA modification sites of the chelator were identified in lysine residues. The biological activity of the conjugates was evaluated by flow cytometry (FACS) using CAIX negative (SKRC-18) and CAIX positive (SKRC-52). The DOTA-G250 conjugates were labelled with 177Lu with a radiochemical yield > 95% reaching specific activities of 12 MBq/µg. The stability in vitro of different types of radioconstructs was analyzed in human serum albumin (HSA). The radiostability of 177Lu-DOTA-G250 at high specific activity was increased by addition of sodium ascorbate after the labelling. The immunoreactivity was evaluated in vitro and in vivo. Binding to CAIX positive cells (SK-RC-52) at different specific activities was higher for conjugates with less DOTA content. Protein dose was optimized in mice with subcutaneously growing SK-RC-52 tumors using different amounts of 177Lu- DOTA-G250.

Keywords: mass spectrometry, monoclonal antibody, radiopharmaceuticals, radioimmunotheray, renal cancer

Procedia PDF Downloads 307