Search results for: biomass protein
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3264

Search results for: biomass protein

2454 Protective Effect of Bexarotene, a Selective RXRα Agonist, against Hypotension Associated with Inflammation and Tissue Injury Linked to Decreased Circulating iNOS Levels in A Rat Model of Septic Shock

Authors: Bahar Tunctan, Sefika Pinar Kucukkavruk, Meryem Temiz-Resitoglu, Demet Sinem Guden, Ayse Nihal Sari, Seyhan Sahan-Firat

Abstract:

We hypothesized that rexinoids such as bexarotene, a selective retinoid X receptor α (RXRα) agonist, may be beneficial for preventing mortality due to inflammation associated with increased expression/activity of inducible nitric oxide synthase (iNOS) induced by lipopolysaccharide (LPS). Therefore, we investigated effects of bexarotene on the changes in circulating protein levels of iNOS (an index for systemic iNOS expression), myeloperoxidase (MPO) (an index for systemic inflammation), and lactate dehydrogenase (LDH) (an index for systemic tissue injury) in LPS-induced systemic inflammation model resulting in septic shock in rats. Rats were injected with saline (4 ml/kg; i.p.), LPS (10 mg/kg; i.p.), dimethylsulphoxide (4 ml/kg, 0.1%; s.c.) at time 0. Mean arterial blood pressure and heart rate were measured using a tail-cuff device. Bexarotene (0.03, 0.1, 0.3, and 1 mg/kg; s.c.) was administered to separate groups of rats 1 h after injection of saline or LPS. The rats were sacrificed 4 h after saline or LPS injection and blood was collected for measurement of serum iNOS, MPO, and LDH protein levels. Blood pressure decreased by 31 mmHg and heart rate increased by 63 bpm in the LPS-treated rats. Bexarotene at 0.3 and 1 mg/kg doses caused 20% mortality 4 h after LPS injection. In the LPS-treated rats, serum iNOS, MPO, and LDH protein levels were increased. Bexarotene only at 0.1 mg/kg dose prevented the LPS-induced hypotension and increased in iNOS, MPO, and LDH protein levels. These data are consistent with the view that a decrease in systemic iNOS levels contributes to the beneficial effect of bexarotene to prevent the hypotension associated with inflammation and tissue injury during rat endotoxemia. [This work was financially supported by The Scientific and Technological Research Council of Turkey (SBAG-109S121)].

Keywords: bexarotene, inflammation, iNOS, lipopolisaccharide, RXRa

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2453 Mesoporous Titania Thin Films for Gentamicin Delivery and Bone Morphogenetic Protein-2 Immobilization

Authors: Ane Escobar, Paula Angelomé, Mihaela Delcea, Marek Grzelczak, Sergio Enrique Moya

Abstract:

The antibacterial capacity of bone-anchoring implants can be improved by the use of antibiotics that can be delivered to the media after the surgery. Mesoporous films have shown great potential in drug delivery for orthopedic applications, since pore size and thickness can be tuned to produce different surface area and free volume inside the material. This work shows the synthesis of mesoporous titania films (MTF) by sol-gel chemistry and evaporation-induced self-assembly (EISA) on top of glass substrates. Pores with a diameter of 12nm were observed by Transmission Electron Microscopy (TEM). A film thickness of 100 nm was measured by Scanning Electron Microscopy (SEM). Gentamicin was used to study the antibiotic delivery from the film by means of High-performance liquid chromatography (HPLC). The Staphilococcus aureus strand was used to evaluate the effectiveness of the penicillin loaded films toward inhibiting bacterial colonization. MC3T3-E1 pre-osteoblast cell proliferation experiments proved that MTFs have a good biocompatibility and are a suitable surface for MC3T3-E1 cell proliferation. Moreover, images taken by Confocal Fluorescence Microscopy using labeled vinculin, showed good adhesion of the MC3T3-E1 cells to the MTFs, as well as complex actin filaments arrangement. In order to improve cell proliferation Bone Morphogenetic Protein-2 (BMP-2) was adsorbed on top of the mesoporous film. The deposition of the protein was proved by measurements in the contact angle, showing an increment in the hydrophobicity while the protein concentration is higher. By measuring the dehydrogenase activity in MC3T3-E1 cells cultured in dually functionalized mesoporous titatina films with gentamicin and BMP-2 is possible to find an improvement in cell proliferation. For this purpose, the absorption of a yellow-color formazan dye, product of a water-soluble salt (WST-8) reduction by the dehydrogenases, is measured. In summary, this study proves that by means of the surface modification of MTFs with proteins and loading of gentamicin is possible to achieve an antibacterial effect and a cell growth improvement.

Keywords: antibacterial, biocompatibility, bone morphogenetic protein-2, cell proliferation, gentamicin, implants, mesoporous titania films, osteoblasts

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2452 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

Abstract:

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

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2451 Ultrasonic Agglomeration of Protein Matrices and Its Effect on Thermophysical, Macro- and Microstructural Properties

Authors: Daniela Rivera-Tobar Mario Perez-Won, Roberto Lemus-Mondaca, Gipsy Tabilo-Munizaga

Abstract:

Different dietary trends worldwide seek to consume foods with anti-inflammatory properties, rich in antioxidants, proteins, and unsaturated fatty acids that lead to better metabolic, intestinal, mental, and cardiac health. In this sense, food matrices with high protein content based on macro and microalgae are an excellent alternative to meet the new needs of consumers. An emerging and environmentally friendly technology for producing protein matrices is ultrasonic agglomeration. It consists of the formation of permanent bonds between particles, improving the agglomeration of the matrix compared to conventionally agglomerated products (compression). Among the advantages of this process are the reduction of nutrient loss and the avoidance of binding agents. The objective of this research was to optimize the ultrasonic agglomeration process in matrices composed of Spirulina (Arthrospira platensis) powder and Cochayuyo (Durvillae Antartica) flour, by means of the response variable (Young's modulus) and the independent variables were the process conditions (percentage of ultrasonic amplitude: 70, 80 and 90; ultrasonic agglomeration times and cycles: 20, 25 and 30 seconds, and 3, 4 and 5). It was evaluated using a central composite design and analyzed using response surface methodology. In addition, the effects of agglomeration on thermophysical and microstructural properties were evaluated. It was determined that ultrasonic compression with 80 and 90% amplitude caused conformational changes according to Fourier infrared spectroscopy (FTIR) analysis, the best condition with respect to observed microstructure images (SEM) and differential scanning calorimetry (DSC) analysis, was the condition of 90% amplitude 25 and 30 seconds with 3 and 4 cycles of ultrasound. In conclusion, the agglomerated matrices present good macro and microstructural properties which would allow the design of food systems with better nutritional and functional properties.

Keywords: ultrasonic agglomeration, physical properties of food, protein matrices, macro and microalgae

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2450 Efficacy of Pisum sativum and Arbuscular Mycorrhizal Symbiosis for Phytoextraction of Heavy Metalloids from Soil

Authors: Ritu Chaturvedi, Manoj Paul

Abstract:

A pot experiment was conducted to investigate the effect of Arbuscular mycorrhizal fungus (AMF) on metal(loid) uptake and accumulation efficiency of Pisum sativum along with physiological and biochemical response. Plants were grown in soil spiked with 50 and 100 mg kg-1 Pb, 25 and 50 mg kg-1 Cd, 50 and 100 mg kg-1 As and a combination of all three metal(loid)s. A parallel set was maintained and inoculated with arbuscular mycorrhizal fungus for comparison. After 60 days, plants were harvested and analysed for metal(loid) content. A steady increase in metal(loid) accumulation was observed on increment of metal(loid) dose and also on AMF inoculation. Plant height, biomass, chlorophyll, carotenoid and carbohydrate content reduced upon metal(loid) exposure. Increase in enzymatic (CAT, SOD and APX) and nonenzymatic (Proline) defence proteins was observed on metal(loid) exposure. AMF inoculation leads to an increase in plant height, biomass, chlorophyll, carotenoids, carbohydrate and enzymatic defence proteins (p≤0.001) under study; whereas proline content was reduced. Considering the accumulation efficiency and adaptive response of plants and alleviation of stress by AMF, this symbiosis can be applied for on-site remediation of Pb and Cd contaminated soil.

Keywords: heavy metal, mycorrhiza, pea, phyroremediation

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2449 Ascidian Styela rustica Proteins’ Structural Domains Predicted to Participate in the Tunic Formation

Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

Abstract:

Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

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2448 The Impact of Human Intervention on Net Primary Productivity for the South-Central Zone of Chile

Authors: Yannay Casas-Ledon, Cinthya A. Andrade, Camila E. Salazar, Mauricio Aguayo

Abstract:

The sustainable management of available natural resources is a crucial question for policy-makers, economists, and the research community. Among several, land constitutes one of the most critical resources, which is being intensively appropriated by human activities producing ecological stresses and reducing ecosystem services. In this context, net primary production (NPP) has been considered as a feasible proxy indicator for estimating the impacts of human interventions on land-uses intensity. Accordingly, the human appropriation of NPP (HANPP) was calculated for the south-central regions of Chile between 2007 and 2014. The HANPP was defined as the difference between the potential NPP of the naturally produced vegetation (NPP0, i.e., the vegetation that would exist without any human interferences) and the NPP remaining in the field after harvest (NPPeco), expressed in gC/m² yr. Other NPP flows taken into account in HANPP estimation were the harvested (NPPh) and the losses of NPP through land conversion (NPPluc). The ArcGIS 10.4 software was used for assessing the spatial and temporal HANPP changes. The differentiation of HANPP as % of NPP0 was estimated by each landcover type taken in 2007 and 2014 as the reference years. The spatial results depicted a negative impact on land use efficiency during 2007 and 2014, showing negative HANPP changes for the whole region. The harvest and biomass losses through land conversion components are the leading causes of loss of land-use efficiency. Furthermore, the study depicted higher HANPP in 2014 than in 2007, representing 50% of NPP0 for all landcover classes concerning 2007. This performance was mainly related to the higher volume of harvested biomass for agriculture. In consequence, the cropland depicted the high HANPP followed by plantation. This performance highlights the strong positive correlation between the economic activities developed into the region. This finding constitutes the base for a better understanding of the main driving force influencing biomass productivity and a powerful metric for supporting the sustainable management of land use.

Keywords: human appropriation, land-use changes, land-use impact, net primary productivity

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2447 Sequence Analysis and Structural Implications of Rotavirus Capsid Proteins

Authors: Nishal Parbhoo, John B. Dewar, Samantha Gildenhuys

Abstract:

Rotavirus is the major cause of severe gastroenteritis worldwide in children aged 5 and younger. Death rates are high particularly in developing countries. The mature rotavirus is a non-enveloped triple-layered nucleocapsid containing 11 double-stranded RNA segments. Here a global view on the sequence and structure of the three main capsid proteins, VP7, VP6, and VP2 is taken by generating a consensus sequence for each of these rotavirus proteins, for each species obtained from published data of representative rotavirus genotypes from across the world and across species. The degree of conservation between species was represented on homology models for each of the proteins. VP7 shows the highest level of variation with 14 - 45 amino acids showing conservation of less than 60%. These changes are localized to the outer surface which is exposed to antibodies alluding to a possible mechanism in evading the immune system. The middle layer, VP6 shows lower variability with only 14-32 sites having lower than 70% conservation. The inner structural layer made up of VP2 showed the lowest variability with only 1-16 sites having less than 70% conservation across species. The results correlate with proteins’ multiple structural roles. Although the nucleotide sequences vary due to an error-prone replication and lack of proofreading, the corresponding amino acid sequence of VP2, 6 and 7 remains conserved. Sequence conservation maintained for the virus results in stable protein structures, fit for function. This can be exploited in drug design, molecular studies and biotechnological applications.

Keywords: amino acid sequence conservation, capsid protein, protein structure, vaccine candidate

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2446 Genetic Variation of Lactoferrin Gene and Its Association with Productive Traits in Egyptian Goats

Authors: Othman E. Othman, Hassan R. Darwish, Amira M. Nowier

Abstract:

Lactoferrin (LF) is a multifunctional protein involved in economically production traits like milk protein composition and skeletal structure in small ruminants including sheep and goat. So, LF gene - with its genetic polymorphisms associated with production traits - is considered a candidate genetic marker used in marker-assisted selection in goats. This study aimed to identify the different alleles and genotypes of this gene in three Egyptian goat breeds using PCR-SSCP (polymerase chain reaction-single-strand conformation polymorphism) and DNA sequencing. Genomic DNA was extracted from 120 animals belonging to Barki, Zaraibi, and Damascus goat breeds. Using specific primers, PCR amplified 247-bp fragments from exon 2 of LF goat gene. The PCR products were subjected to Single-Strand Conformation Polymorphism (SSCP) technique. The results showed the presence of two genotypes GG and AG in the tested animals. The frequencies of both genotypes varied among the three tested breeds with the highest frequencies of GG genotype in all tested goat breeds. The sequence analysis of PCR products representing these two detected genotypes declared the presence of an SNP (single nucleotide polymorphisms) substitution (G/A) among G and A alleles of this gene. The association between different LF genotypes and milk composition as well as body measurement was estimated. The comparison showed that the animals possess AG genotypes are superior over those with GG genotypes for different parameters of milk protein compositions and skeletal structures. This finding declared that allele A of LF gene is considered the promising marker for the productive traits in goat. In conclusion, the Egyptian goat breeds will be needed to enhance their milk protein composition and growth trait parameters through the increasing of allele A frequency in their herds depending on the superior production traits of this allele in goats.

Keywords: lLactoferrin gene, PCR-SSCP, SNPs, Egyptian goat

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2445 LHCII Proteins Phosphorylation Changes Involved in the Dark-Chilling Response in Plant Species with Different Chilling Tolerance

Authors: Malgorzata Krysiak, Anna Wegrzyn, Maciej Garstka, Radoslaw Mazur

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Under constantly fluctuating environmental conditions, the thylakoid membrane protein network evolved the ability to dynamically respond to changing biotic and abiotic factors. One of the most important protective mechanism is rearrangement of the chlorophyll-protein (CP) complexes, induced by protein phosphorylation. In a temperate climate, low temperature is one of the abiotic stresses that heavily affect plant growth and productivity. The aim of this study was to determine the role of LHCII antenna complex phosphorylation in the dark-chilling response. The study included an experimental model based on dark-chilling at 4 °C of detached chilling sensitive (CS) runner bean (Phaseolus coccineus L.) and chilling tolerant (CT) garden pea (Pisum sativum L.) leaves. This model is well described in the literature as used for the analysis of chilling impact without any additional effects caused by light. We examined changes in thylakoid membrane protein phosphorylation, interactions between phosphorylated LHCII (P-LHCII) and CP complexes, and their impact on the dynamics of photosystem II (PSII) under dark-chilling conditions. Our results showed that the dark-chilling treatment of CS bean leaves induced a substantial increase of phosphorylation of LHCII proteins, as well as changes in CP complexes composition and their interaction with P-LHCII. The PSII photochemical efficiency measurements showed that in bean, PSII is overloaded with light energy, which is not compensated by CP complexes rearrangements. On the contrary, no significant changes in PSII photochemical efficiency, phosphorylation pattern and CP complexes interactions were observed in CT pea. In conclusion, our results indicate that different responses of the LHCII phosphorylation to chilling stress take place in CT and CS plants, and that kinetics of LHCII phosphorylation and interactions of P-LHCII with photosynthetic complexes may be crucial to chilling stress response. Acknowledgments: presented work was financed by the National Science Centre, Poland grant No.: 2016/23/D/NZ3/01276

Keywords: LHCII, phosphorylation, chilling stress, pea, runner bean

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2444 A Review of Energy in the Democratic Republic of Congo

Authors: Kanzumba Kusakana

Abstract:

The Democratic Republic of Congo (DRC) is currently experiencing a general energy crisis due to lack of proper investment and management in the energy sector. 93, 6% of the country is highly dependent on wood fuels as main source of energy having severe impacts such as deforestation and general degradation of the environment. On the other hand, the major share of the electricity produced mainly from ill-conditioned hydro and thermal power stations is principally used to supply the industrial sector as well as very few urban areas. Nevertheless, DRC possesses huge potential in renewable resources such as hydropower, biomass, methane gas, solar geothermal and moderate wind potential that can be used for energy generation. Recently the country has initiated projects to build decentralized micro hydropower station to supply remotes and isolated areas; to rehabilitate its existent main hydropower plants and transmission lines as well as to extend its current generation capacity by building new hydropower stations able to respond to a major part of the African continent energy needs. This paper presents a comprehensive review of current energy resources as well as of the electricity situation in DRC. Recent energy projects, the energy policy as well as the energy challenges in the DRC are also presented.

Keywords: energy, biomass, hydro power, renewable energy, energy policy, Democratic Republic of Congo

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2443 Intentional Cultivation of Non-toxic Filamentous Cyanobacteria Tolypothrix as an Approach to Treat Eutrophic Waters

Authors: Simona Lucakova, Irena Branyikova

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Eutrophication, a condition when water becomes over-enriched with nutrients (P, N), can lead to undesirable excessive growth of phytoplankton, so-called algal bloom. This process results in the accumulation of toxin-producing cyanobacteria and oxygen depletion, both possibly leading to the collapse of the whole ecosystem. In real conditions, the limiting nutrient, which determines the possible growth of harmful algal bloom, is usually phosphorus. Algicides or flocculants have been applied in the eutrophicated waterbody in order to reduce the phytoplankton growth, which leads to the introduction of toxic chemicals into the water. In our laboratory, the idea of the prevention of harmful phytoplankton growth by the intentional cultivation of non-toxic cyanobacteria Tolypothrix tenuis in semi-open floating photobioreactors directly on the surface of phosphorus-rich waterbody is examined. During the process of cultivation, redundant phosphorus is incorporated into cyanobacterial biomass, which can be subsequently used for the production of biofuels, cosmetics, pharmaceuticals, or biostimulants for agricultural use. To determine the ability of phosphorus incorporation, batch-cultivation of Tolypothrix biomass in media simulating eutrophic water (10% BG medium) and in effluent from municipal wastewater treatment plant, both with the initial phosphorus concentration in the range 0.5-1.0 mgP/L was performed in laboratory-scale models of floating photobioreactors. After few hours of cultivation, the phosphorus content was decreased below the target limit of 0.035 mgP/L, which was given as a borderline for the algal bloom formation. Under laboratory conditions, the effect of several parameters on the rate of phosphorus decrease was tested (illumination, temperature, stirring speed/aeration gas flow, biomass to medium ratio). Based on the obtained results, a bench-scale floating photobioreactor was designed and will be tested for Tolypothrix growth in real conditions. It was proved that intentional cultivation of cyanobacteria Tolypothrix could be a suitable approach for extracting redundant phosphorus from eutrophic waters as prevention of algal bloom formation.

Keywords: cyanobacteria, eutrophication, floating photobioreactor, Tolypothrix

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2442 Fabrication of Electrospun Green Fluorescent Protein Nano-Fibers for Biomedical Applications

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

Abstract:

GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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2441 Detection of Heroin and Its Metabolites in Urine Samples: A Chemiluminescence Approach

Authors: Sonu Gandhi, Neena Capalash, Prince Sharma, C. Raman Suri

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A sensitive chemiluminescence immunoassay (CIA) for heroin and its major metabolites is reported. The method is based on the competitive reaction of horseradish peroxidase (HRP)-labeled anti-MAM antibody and free drug in spiked urine samples. A hapten-protein conjugate was synthesized by using acidic derivative of monoacetyl morphine (MAM) coupled to carrier protein BSA and was used as an immunogen for the generation of anti-MAM (monoacetyl morphine) antibody. A high titer of antibody (1:64,0000) was obtained and the relative affinity constant (Kaff) of antibody was 3.1×107 l/mol. Under the optimal conditions, linear range and reactivity for heroin, mono acetyl morphine (MAM), morphine and codeine were 0.08, 0.09, 0.095 and 0.092 ng/mL respectively. The developed chemiluminescence inhibition assay could detect heroin and its metabolites in standard and urine samples up to 0.01 ng/ml.

Keywords: heroin, metabolites, chemiluminescence immunoassay, horse radish peroxidase

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2440 Ecosystem Response of a Semi-Enclosed Saline Bay to Damming and Sluice-Management: Case of Lake Grevelingen in the Netherlands

Authors: Marijn Tangelder, Ingeborg Mulder, Jeroen Wijsman, John Janssen, Tom Ysebaert

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The delta works in the Southwest Netherlands drastically changed the estuarine nature of this area. The Grevelingen estuary was dammed off and became a lake in 1971. Since 1978 a connection with the North Sea exists to keep the lake saline but management of the sluices varied over time. Our research of several decades of monitoring data shows that water management practices lead to drastic changes in water quality and consequent ecological shifts in benthic fauna, fish, and bird species. Benthic biomass, dominated by molluscs, showed major changes with an important role for invasive species. Fish migration and, therefore, fish stock in the lake changed with recently smaller fish species and lower biomass values, with consequences for fish eating birds. Implications are made towards future management to re-introduce micro-tide in connection with the North Sea to improve water quality and the ecological status of the lake, as well as consequences for the bordering Natura 2000 terrestrial habitats, including rare dune vegetations, are discussed.

Keywords: ecosystem study, Grevelingen, Natura 2000, water management, water quality

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2439 Using Lysosomal Immunogenic Cell Death to Target Breast Cancer via Xanthine Oxidase/Micro-Antibody Fusion Protein

Authors: Iulianna Taritsa, Kuldeep Neote, Eric Fossel

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Lysosome-induced immunogenic cell death (LIICD) is a powerful mechanism of targeting cancer cells that kills circulating malignant cells and primes the host’s immune cells against future remission. Current immunotherapies for cancer are limited in preventing recurrence – a gap that can be bridged by training the immune system to recognize cancer neoantigens. Lysosomal leakage can be induced therapeutically to traffic antigens from dying cells to dendritic cells, which can later present those tumorigenic antigens to T cells. Previous research has shown that oxidative agents administered in the tumor microenvironment can initiate LIICD. We generated a fusion protein between an oxidative agent known as xanthine oxidase (XO) and a mini-antibody specific for EGFR/HER2-sensitive breast tumor cells. The anti-EGFR single domain antibody fragment is uniquely sourced from llama, which is functional without the presence of a light chain. These llama micro-antibodies have been shown to be better able to penetrate tissues and have improved physicochemical stability as compared to traditional monoclonal antibodies. We demonstrate that the fusion protein created is stable and can induce early markers of immunogenic cell death in an in vitro human breast cancer cell line (SkBr3). Specifically, we measured overall cell death, as well as surface-expressed calreticulin, extracellular ATP release, and HMGB1 production. These markers are consensus indicators of ICD. Flow cytometry, luminescence assays, and ELISA were used respectively to quantify biomarker levels between treated versus untreated cells. We also included a positive control group of SkBr3 cells dosed with doxorubicin (a known inducer of LIICD) and a negative control dosed with cisplatin (a known inducer of cell death, but not of the immunogenic variety). We looked at each marker at various time points after cancer cells were treated with the XO/antibody fusion protein, doxorubicin, and cisplatin. Upregulated biomarkers after treatment with the fusion protein indicate an immunogenic response. We thus show the potential for this fusion protein to induce an anticancer effect paired with an adaptive immune response against EGFR/HER2+ cells. Our research in human cell lines here provides evidence for the success of the same therapeutic method for patients and serves as the gateway to developing a new treatment approach against breast cancer.

Keywords: apoptosis, breast cancer, immunogenic cell death, lysosome

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2438 Estimations of Spectral Dependence of Tropospheric Aerosol Single Scattering Albedo in Sukhothai, Thailand

Authors: Siriluk Ruangrungrote

Abstract:

Analyses of available data from MFR-7 measurement were performed and discussed on the study of tropospheric aerosol and its consequence in Thailand. Since, ASSA (w) is one of the most important parameters for a determination of aerosol effect on radioactive forcing. Here the estimation of w was directly determined in terms of the ratio of aerosol scattering optical depth to aerosol extinction optical depth (ωscat/ωext) without any utilization of aerosol computer code models. This is of benefit for providing the elimination of uncertainty causing by the modeling assumptions and the estimation of actual aerosol input data. Diurnal w of 5 cloudless-days in winter and early summer at 5 distinct wavelengths of 415, 500, 615, 673 and 870 nm with the consideration of Rayleigh scattering and atmospheric column NO2 and Ozone contents were investigated, respectively. Besides, the tendency of spectral dependence of ω representing two seasons was observed. The characteristic of spectral results reveals that during wintertime the atmosphere of the inland rural vicinity for the period of measurement possibly dominated with a lesser amount of soil dust aerosols loading than one in early summer. Hence, the major aerosol loading particularly in summer was subject to a mixture of both soil dust and biomass burning aerosols.

Keywords: aerosol scattering optical depth, aerosol extinction optical depth, biomass burning aerosol, soil dust aerosol

Procedia PDF Downloads 389
2437 Protein-Thiocyanate Composite as a Sensor for Iron III Cations

Authors: Hosam El-Sayed, Amira Abou El-Kheir, Salwa Mowafi, Marwa Abou Taleb

Abstract:

Two proteinic biopolymers; namely keratin and sericin, were extracted from their respective natural resources by simple appropriate methods. The said proteins were dissolved in the appropriate solvents followed by regeneration in a form of film polyvinyl alcohol. Proteinium thiocyanate (PTC) composite was prepared by reaction of a regenerated film with potassium thiocyanate in acid medium. In another experiment, the said acidified proteins were reacted with potassium thiocyante before dissolution and regeneration in a form of PTC composite. The possibility of using PTC composite for determination of the concentration of iron III ions in domestic as well as industrial water was examined. The concentration of iron III cations in water was determined spectrophotometrically by measuring the intensity of blood red colour of iron III thiocyanate obtained by interaction of PTC with iron III cation in the tested water sample.

Keywords: iron III cations, protein, sensor, thiocyanate, water

Procedia PDF Downloads 409
2436 Effect of Different Commercial Diets and Temperature on the Growth Performance, Feed Intake and Feed Conversion Ratio of Sobaity Seabream Sparidentex hasta

Authors: Seemab Zehra, A. H. W. Mohammed, E. Pantanella, J. L. Q. Laranja, P. H. De Mello, R. Saleh, A. A. Siddik, A. Al Shaikhi, A. M. Al-Suwailem

Abstract:

Two separate feeding trials were conducted to determine the effects of using different commercial diets and water temperatures on the growth performance, feed intake, feed conversion ratio (FCR) and condition factor of sobaity seabream Sparidentex hasta. In experiment I, growth performance, feed intake, protein efficiency ratio (PER), feed conversion ratio (FCR) and survival (%) of sobaity seabream Sparidentex hasta (330.5±2.6 g; 26.9±1.0 cm) were evaluated by four different commercial diets (1, 2, 3 and 4) for 80 days. The daily weight gain was around 3.2 g day-1 with an SGR of 0.7% day-1. Both the FCR and PER in the fish were significantly better in diet 2 that contained 46.36% crude protein and 12.54% crude fat. In experiment II, (99±2.6 g; 17.1±1.0 cm). The fish were cultured in 1m3 tanks supplied with seawater from the Red Sea wherein three different rearing temperatures were set as treatments (24, 28 and 32°C). Fish were fed with a commercial diet based on the results of experiment I (46.4% protein; 20.1 MJ kg-1 energy) to satiation for 96 days. Total weight gain was significantly higher for the fish reared in the 32°C group (158.57 g) followed by the 28°C group (138.25 g), while the lowest weight gain was observed in the 24°C group (116.98 g). The FCR was significantly lower in the 32°C group (1.62) as compared to 28 (1.8) and 24°C (1.85) groups. Based on the results obtained from these preliminary studies (experiment I and II), sobaity seabream can attain better growth performance, FCR and PER at 32°C in the Red Sea by feeding commercial diet 2.

Keywords: Sparidentex hasta, nutrition, FCR, Red Sea, growth performance

Procedia PDF Downloads 60
2435 Pharmacokinetics, Dosage Regimen and in Vitro Plasma Protein Binding of Danofloxacin following Intravenous Administration in Adult Buffaloes

Authors: Zahid Manzoor, Shaukat Hussain Munawar, Zahid Iqbal, Imran Ahmad Khan, Abdul Aziz, Hafiz Muhammad Qasim

Abstract:

The present study was aimed to investigate the pharmacokinetics behavior and optimal dosage regimen of danofloxacin in 8 adult healthy buffaloes of local breed (Nili Ravi) following single intravenous administration at the dose of 2.5 mg/kg body weight. Plasma drug concentrations at various time intervals were measured by HPLC method. In vitro plasma protein binding was determined employing the ultrafiltration technique. The distribution and elimination of danofloxacin was rapid, as indicated by the values (Mean±SD) of distribution half-life (t1/2α = 0.25±0.09 hours) and elimination half life (t1/2β = 3.26±0.43 hours), respectively. Volume of distribution at steady state (Vss) was 1.14±0.12 L/kg, displaying its extensive distribution into various body fluids and tissues. The high value of AUC (9.80±2.14 µg/ml.hr) reflected the vast area of the body covered by drug concentration. The mean residence time was noted to be 4.78±0.52 hours. On the basis of pharmacokinetic parameters, a suitable intravenous regimen for danofloxacin in adult buffaloes would be 6.5 mg/kg to be repeated after 12 hours intervals. The present study is the foremost pharmacokinetic study of danofloxacin in the local species which would provide the valueable contribution in the local manufacturing of danofloxacin in Pakistan in future.

Keywords: danofloxacin, pharmacokinetics, plasma protein binding, buffaloes, dosage regimen

Procedia PDF Downloads 592
2434 Forage Quality of Chickpea - Barley as Affected by Mixed Cropping System in Water Stress Condition

Authors: Masoud Rafiee

Abstract:

To study the quality response of forage to chickpea-barley mixed cropping under drought stress and vermicompost consumption, an experiment was carried out under well watered and %70 water requirement (stress condition) in RCBD as split plot with four replications in temperate condition of Khorramabad in 2013. Chickpea-barley mix cropping (%100 chickpea, %75:25 chickpea:barley, %50:50 chickpea:barley, %25:75 chickpea:barley, and %100 barley) was studied. Results showed that wet and dry forage yield were significantly affected by environment and decreased in stress condition. Also, crude protein content decreased from %26.2 in well watered to %17.3 in stress condition.

Keywords: crude protein, wet forage yield, dry forage yield, water stress condition, well watered

Procedia PDF Downloads 331
2433 Supplementation of Jackfruit By-Product Concentrate in Combination with Two Types of Protein Sources for Growing Kids

Authors: Emely J. Escala, Lolito C. Bestil

Abstract:

An experiment was conducted to assess the potential of jackfruit by-product concentrate (JBC) in combination with two types of protein sources, soybean meal (SBM) and liquid acid whey (LAW), given at two different ratios as supplement for growing kids fed a basal diet of 70:30 napier grass (Pennisetum purpureum) and kakawate (Gliricidia sepium) soilage ratio. The experiment was set-up in randomized complete block design (RCBD) with sex-age combination as basis for blocking, with the following dietary treatments: T1 = 0.50:0.50% BW, DM basis, JBC:SBM, T2 = 0.75:0.25% BW JBC:SBM, T3 = 0.50:0.50% BW, DM basis, JBC:LAW, and T4 = 0.75:0.25% BW JBC:LAW. Analysis of JBC showed high contents of crude fiber with medium levels of crude protein and nitrogen-free extract, appearing to be fitting for ruminants and a potential energy source. Results showed significantly higher voluntary dry matter intake (VDMI), cumulative weight gain (CWG), and average daily gain (ADG) of growing goats supplemented with JBC in combination with SBM than with LAW. The amount of JBC can range from 0.50% to 0.75% BW with SBM making up the difference, but a JBC:SBM ratio of 0.75:0.25% BW, DM basis, is best in promoting highest voluntary dry matter intake and is, therefore, highly recommended in the light of savings in feed cost. A long-term study on the effects of JBC supplementation on meat qualities of growing kids (aroma, marbling characteristics and taste) is also recommended.

Keywords: jackfruit by-product concentrate, liquid acid whey, soybean meal, grower kids

Procedia PDF Downloads 179
2432 Assessment of cellulase and xylanase Production by chryseobacterium sp. Isolated from Decaying Biomass in Alice, Eastern Cape, South Africa

Authors: A. Nkohla, U. Nwodo, L. V. Mabinya, A. I. Okoh

Abstract:

A potential source for low-cost production of value added products is the utilization of lignocellulosic materials. However, the huddle needing breaching would be the dismantlement of the complex lignocellulosic structure as to free sugar base therein. the current lignocellosic material treatment process is expensive and not eco-friendly hence, the advocacy for enzyme based technique which is both cheap and eco-friendly is highly imperative. Consequently, this study aimed at the screening of cellulose and xylan degrading bacterial strain isolated from decaying sawdust samples. This isolate showed high activity for cellulase and xylanase when grown on carboxymethyl cellulose and birtchwood xylan as the sole carbon source respectively. The 16S rDNA nucleotide sequence of the isolate showed 98% similarity with that of Chryseobacterium taichungense thus, it was identified as a Chryseobacterium sp. Optimum culture conditions for cellulase and xylanase production were medium pH 6, incubation temperature of 25 °C at 50 rpm and medium pH 6, incubation temperature of 25 °C at 150 rpm respectively. The high enzyme activity obtained from this bacterial strain portends it as a good candidate for industrial use in the degradation of complex biomass for value added products.

Keywords: lignocellulosic material, chryseobacterium sp., submerged fermentation, cellulase, xylanase

Procedia PDF Downloads 292
2431 IL6/PI3K/mTOR/GFAP Molecular Pathway Role in COVID-19-Induced Neurodegenerative Autophagy, Impacts and Relatives

Authors: Mohammadjavad Sotoudeheian

Abstract:

COVID-19, which began in December 2019, uses the angiotensin-converting enzyme 2 (ACE2) receptor to enter and spread through the cells. ACE2 mRNA is present in almost every organ, including nasopharynx, lung, as well as the brain. Ports of entry of SARS-CoV-2 into the central nervous system (CNS) may include arterial circulation, while viremia is remarkable. However, it is imperious to develop neurological symptoms evaluation CSF analysis in patients with COVID-19, but theoretically, ACE2 receptors are expressed in cerebellar cells and may be a target for SARS-CoV-2 infection in the brain. Recent evidence agrees that SARS-CoV-2 can impact the brain through direct and indirect injury. Two biomarkers for CNS injury, glial fibrillary acidic protein (GFAP) and neurofilament light chain (NFL) detected in the plasma of patients with COVID-19. NFL, an axonal protein expressed in neurons, is related to axonal neurodegeneration, and GFAP is over-expressed in CNS inflammation. GFAP cytoplasmic accumulation causes Schwan cells to misfunction, so affects myelin generation, reduces neuroskeletal support over NfLs during CNS inflammation, and leads to axonal degeneration. Interleukin-6 (IL-6), which extensively over-express due to interleukin storm during COVID-19 inflammation, regulates gene expression, as well as GFAP through STAT molecular pathway. IL-6 also impresses the phosphoinositide 3-kinase (PI3K)/STAT/smads pathway. The PI3K/ protein kinase B (Akt) pathway is the main modulator upstream of the mammalian target of rapamycin (mTOR), and alterations in this pathway are common in neurodegenerative diseases. Most neurodegenerative diseases show a disruption of autophagic function and display an abnormal increase in protein aggregation that promotes cellular death. Therefore, induction of autophagy has been recommended as a rational approach to help neurons clear abnormal protein aggregates and survive. The mTOR is a major regulator of the autophagic process and is regulated by cellular stressors. The mTORC1 pathway and mTORC2, as complementary and important elements in mTORC1 signaling, have become relevant in the regulation of the autophagic process and cellular survival through the extracellular signal-regulated kinase (ERK) pathway.

Keywords: mTORC1, COVID-19, PI3K, autophagy, neurodegeneration

Procedia PDF Downloads 74
2430 Mechanisms Leading to the Protective Behavior of Ethanol Vapour Drying of Probiotics

Authors: Shahnaz Mansouri, Xiao Dong Chen, Meng Wai Woo

Abstract:

A new antisolvent vapour precipitation approach was used to make ultrafine submicron probiotic encapsulates. The approach uses ethanol vapour to precipitate submicron encapsulates within relatively large droplets. Surprisingly, the probiotics (Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus) showed relatively high survival even under destructive ethanolic conditions within the droplet. This unusual behaviour was deduced to be caused by the denaturation and aggregation of the milk protein forming an ethanolic protective matrix for the probiotics. Skim milk droplets which is rich in casein and contains naturally occurring minerals provided higher ethanolic protection when compared whey protein isolate and lactose droplets.

Keywords: whey, skim milk, probiotic, antisolvent, precipitation, encapsulation, denaturation, aggregation

Procedia PDF Downloads 509
2429 Inulinase Immobilization on Functionalized Magnetic Nanoparticles Prepared with Soy Protein Isolate Conjugated Bovine Serum Albumin for High Fructose Syrup Production

Authors: Homa Torabizadeh, Mohaddeseh Mikani

Abstract:

Inulinase from Aspergillus niger was covalently immobilized on magnetic nanoparticles (MNPs/Fe3O4) covered with soy protein isolate (SPI/Fe3O4) functionalized by bovine serum albumin (BSA) nanoparticles. MNPs are promising enzyme carriers because they separate easily under external magnetic fields and have enhanced immobilized enzyme reusability. As MNPs aggregate simply, surface coating strategy was employed. SPI functionalized by BSA was a suitable candidate for nanomagnetite coating due to its superior biocompatibility and hydrophilicity. Fe3O4@SPI-BSA nanoparticles were synthesized as a novel carrier with narrow particle size distribution. Step by step fabrication monitoring of Fe3O4@SPI-BSA nanoparticles was performed using field emission scanning electron microscopy and dynamic light scattering. The results illustrated that nanomagnetite with the spherical morphology was well monodispersed with the diameter of about 35 nm. The average size of the SPI-BSA nanoparticles was 80 to 90 nm, and their zeta potential was around −34 mV. Finally, the mean diameter of fabricated Fe3O4@SPI-BSA NPs was less than 120 nm. Inulinase enzyme from Aspergillus niger was covalently immobilized through gluteraldehyde on Fe3O4@SPI-BSA nanoparticles successfully. Fourier transform infrared spectra and field emission scanning electron microscopy images provided sufficient proof for the enzyme immobilization on the nanoparticles with 80% enzyme loading.

Keywords: high fructose syrup, inulinase immobilization, functionalized magnetic nanoparticles, soy protein isolate

Procedia PDF Downloads 279
2428 Investigation of a Single Feedstock Particle during Pyrolysis in Fluidized Bed Reactors via X-Ray Imaging Technique

Authors: Stefano Iannello, Massimiliano Materazzi

Abstract:

Fluidized bed reactor technologies are one of the most valuable pathways for thermochemical conversions of biogenic fuels due to their good operating flexibility. Nevertheless, there are still issues related to the mixing and separation of heterogeneous phases during operation with highly volatile feedstocks, including biomass and waste. At high temperatures, the volatile content of the feedstock is released in the form of the so-called endogenous bubbles, which generally exert a “lift” effect on the particle itself by dragging it up to the bed surface. Such phenomenon leads to high release of volatile matter into the freeboard and limited mass and heat transfer with particles of the bed inventory. The aim of this work is to get a better understanding of the behaviour of a single reacting particle in a hot fluidized bed reactor during the devolatilization stage. The analysis has been undertaken at different fluidization regimes and temperatures to closely mirror the operating conditions of waste-to-energy processes. Beechwood and polypropylene particles were used to resemble the biomass and plastic fractions present in waste materials, respectively. The non-invasive X-ray technique was coupled to particle tracking algorithms to characterize the motion of a single feedstock particle during the devolatilization with high resolution. A high-energy X-ray beam passes through the vessel where absorption occurs, depending on the distribution and amount of solids and fluids along the beam path. A high-speed video camera is synchronised to the beam and provides frame-by-frame imaging of the flow patterns of fluids and solids within the fluidized bed up to 72 fps (frames per second). A comprehensive mathematical model has been developed in order to validate the experimental results. Beech wood and polypropylene particles have shown a very different dynamic behaviour during the pyrolysis stage. When the feedstock is fed from the bottom, the plastic material tends to spend more time within the bed than the biomass. This behaviour can be attributed to the presence of the endogenous bubbles, which drag effect is more pronounced during the devolatilization of biomass, resulting in a lower residence time of the particle within the bed. At the typical operating temperatures of thermochemical conversions, the synthetic polymer softens and melts, and the bed particles attach on its outer surface, generating a wet plastic-sand agglomerate. Consequently, this additional layer of sand may hinder the rapid evolution of volatiles in the form of endogenous bubbles, and therefore the establishment of a poor drag effect acting on the feedstock itself. Information about the mixing and segregation of solid feedstock is of prime importance for the design and development of more efficient industrial-scale operations.

Keywords: fluidized bed, pyrolysis, waste feedstock, X-ray

Procedia PDF Downloads 155
2427 Studies on Some Aspects of Sub Clinical Mastitis in Cattle

Authors: Kavita Jaidiya, Anju Chahar, Chitra Jaidiya

Abstract:

The present study was conducted on 200 quarters from 50 apparently healthy cows. Samples are subjected to California Mastitis Test (CMT), cultural examination, and mPCR. Milk samples were also subjected to changes in composition Viz. fat, protein, and lactose. The prevalence of subclinical mastitis based on culture examination was 30(60/200), 36 (72/200), and 40 percent (93/200) based on CMT, culture examination, and mPCR on a quarterly basis. The prevalence of subclinical mastitis on animal basis was 40 (20/50), 46 (23/50), and 52 percent (26/50) based on CMT, Culture examination, and mPCR. The highest prevalence was observed in IVth parity on a quarterly basis and in Vth parity on cow basis. On culture examination, Staphylococcus aureus was the most prevalent organism (50.56%), followed by Streptococcus dysaglactiae (11.33%), E. coli (7.8 %), Staphylococcus agalactiae (13.48 %), Staphylococcus epidermidis (2.2 %), Streptococcus hyicus (6.94%), Streptococcus uberis (5.16%), Klebsiella pneumonia (6.74%). On isolation by bacterial mPCR, Staphylococcus spp. (42%) was the major pathogen. Organisms isolated in mixed infections are Streptococcus spp., Klebsiella pneumonia, E.coli and Pseudomonas aeruginous. The average mean value of fat, protein, and lactose content in subclinically affected milk samples were 3.40 ± 0.101, 3.009 ± 0.033, and 4.48 ± 0.03, and the mean value of fat, protein, and lactose content in normal milk were 4.13 ± 0.035, 3.39 ± 0.021, and 5.10 ± 0.016. The mean blood level of reduced glutathione in subclinical mastitis (30.44 ± 1.87 ng/ml) was lower than healthy cows (47.98 ± 4.04ng/ml). The concentration of malondialdehyde (10.026 ± 0.21mmol/L) in subclinical mastitis was significantly higher as compared to healthy group cows (2.19 ± 0.23mmol/L).

Keywords: cow, subclinical mastitis, mPCR, California Mastitis test

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2426 Isosorbide Bis-Methyl Carbonate: Opportunities for an Industrial Model Based on Biomass

Authors: Olga Gomez De Miranda, Jose R. Ochoa-Gomez, Stefaan De Wildeman, Luciano Monsegue, Soraya Prieto, Leire Lorenzo, Cristina Dineiro

Abstract:

The chemical industry is facing a new revolution. As long as processes based on the exploitation of fossil resources emerged with force in the XIX century, Society currently demands a new radical change that will lead to the complete and irreversible implementation of a circular sustainable economic model. The implementation of biorefineries will be essential for this. There, renewable raw materials as sugars and other biomass resources are exploited for the development of new materials that will partially replace their petroleum-derived homologs in a safer, and environmentally more benign approach. Isosorbide, (1,4:3,6-dianhydro-d-glucidol) is a primary bio-based derivative obtained from the plant (poly) saccharides and a very interesting example of a useful chemical produced in biorefineries. It can, in turn, be converted to other secondary monomers as isosorbide bis-methyl carbonate (IBMC), whose main field of application can be as a key biodegradable intermediary substitute of bisphenol-A in the manufacture of polycarbonates, or as an alternative to the toxic isocyanates in the synthesis of new polyurethanes (non-isocyanate polyurethanes) both with a huge application market. New products will present advantageous mechanical or optical properties, as well as improved behavior in non-toxicity and biodegradability aspects in comparison to their petro-derived alternatives. A robust production process of IBMC, a biomass-derived chemical, is here presented. It can be used with different raw material qualities using dimethyl carbonate (DMC) as both co-reactant and solvent. It consists of the transesterification of isosorbide with DMC under soft operational conditions, using different basic catalysts, always active with the isosorbide characteristics and purity. Appropriate isolation processes have been also developed to obtain crude IBMC yields higher than 90%, with oligomers production lower than 10%, independently of the quality of the isosorbide considered. All of them are suitable to be used in polycondensation reactions for polymers obtaining. If higher qualities of IBMC are needed, a purification treatment based on nanofiltration membranes has been also developed. The IBMC reaction-isolation conditions established in the laboratory have been successfully modeled using appropriate software programs and moved to a pilot-scale (production of 100 kg of IBMC). It has been demonstrated that a highly efficient IBMC production process able to be up-scaled under suitable market conditions has been obtained. Operational conditions involved the production of IBMC involve soft temperature and energy needs, no additional solvents, and high operational efficiency. All of them are according to green manufacturing rules.

Keywords: biomass, catalyst, isosorbide bis-methyl carbonate, polycarbonate, polyurethane, transesterification

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2425 Docking and Dynamic Molecular Study of Isoniazid Derivatives as Anti-Tuberculosis Drug Candidate

Authors: Richa Mardianingrum, Srie R. N. Endah

Abstract:

In this research, we have designed four isoniazid derivatives i.e., isonicotinohydrazide (1-isonicotinoyl semicarbazide, 1-thiosemi isonicotinoyl carbazide, N '-(1,3-dimethyl-1 h-pyrazole-5-carbonyl) isonicotino hydrazide, and N '-(1,2,3- 4-thiadiazole-carbonyl) isonicotinohydrazide. The docking and molecular dynamic have performed to them in order to study its interaction with Mycobacterium tuberculosis Enoyl-Acyl Carrier Protein Reductase (InhA). Based on this research, all of the compounds were predicted to have a stable interaction with Mycobacterium tuberculosis Enoyl-Acyl Carrier Protein Reductase (INHA) receptor, so they could be used as an anti-tuberculosis drug candidate.

Keywords: anti-tuberculosis, docking, Inhibin alpha subunit, InhA, inhibition, synthesis, isonicotinohydrazide

Procedia PDF Downloads 165