Search results for: high cell density cultivation
23819 Production of Recombinant VP2 Protein of Canine Parvovirus Type 2c Using Baculovirus Expression System
Authors: Jae Young Song, In-Ohk Ouh, Seyeon Park, Byeong Sul Kang, Soo Dong Cho, In-Soo Cho
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Canine parvovirus (CPV) is a major pathogen of diarrhea disease in dogs. CPV type 2 has three of antigenic variants such as 2a, 2b, and 2c. CPV constructs a small non-enveloped, icosahedral capsid that contains single-stranded DNA. It has capsids that two largely overlapping virion proteins (VP), VP1 (82 kDa), and VP2 (65 kDa). Baculoviruses are insect pathogens that regulate insect populations in nature and are being successfully used to control insect pests. The proteins produced in the baculovirus-expression system are used for instance for functional studies, vaccine preparations, or diagnostics. The vaccines produced by baculovirus-expression system showed elicitation of antibodies. The recombinant baculovirus infected SF9 cells showed broken shape. The recombinant VP2 proteins from cell pellet or supernatant were confirmed by western blotting. The result showed that the recombinant VP2 protein bands were appeared at 65 kDa molecular weight in both cell pellet and supernatant of infected SF9 cell. These results indicated that the recombinant baculovirus infected SF9 cell express the recombinant VP2 protein successfully. In addition, the expressed recombinant VP2 protein is secreted from cell to supernatant. The baculovirus expression system can be used to produce the VP2 protein of CPV 2c. In addition, the secretion property of the expression of VP2 protein may decrease the cost of production, because it can be skipped the cell breaking step. The produced VP2 protein could be used for vaccine and the agent of diagnostic tests. This study provides the foundation of the production of CPV 2c vaccine and the diagnostic agent.Keywords: baculovirus, canine parvovirus 2c, dog, Korea
Procedia PDF Downloads 15123818 Zinc Oxide Nanorods Decorated Nanofibers Based Flexible Electrodes for Capacitive Energy Storage Applications
Authors: Syed Kamran Sami, Saqib Siddiqui
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In recent times, flexible supercapacitors retaining high electrochemical performance and steadiness along with mechanical endurance has developed as a spring of attraction due to the exponential progress and innovations in energy storage devices. To meet the rampant increasing demand of energy storage device with the small form factor, a unique, low cost and high-performance supercapacitor with considerably higher capacitance and mechanical robustness is required to recognize their real-life applications. Here in this report, synthesis route of electrode materials with low rigidity and high charge storage performance is reported using 1D-1D hybrid structure of zinc oxide (ZnO) nanorods, and conductive polymer smeared polyvinylidene fluoride–trifluoroethylene (P(VDF–TrFE)) electrospun nanofibers. The ZnO nanorods were uniformly grown on poly (3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT: PSS) coated P(VDF-TrFE) nanofibers using hydrothermal growth to manufacture light weight, permeable electrodes for supercapacitor. The PEDOT: PSS coated P(VDF-TrFE) porous web of nanofibers act as framework with high surface area. The incorporation of ZnO nanorods further boost the specific capacitance by 59%. The symmetric device using the fabricated 1D-1D hybrid electrodes reveals fairly high areal capacitance of 1.22mF/cm² at a current density of 0.1 mA/cm² with a power density of more than 1600 W/Kg. Moreover, the fabricated electrodes show exceptional flexibility and high endurance with 90% and 76% specific capacitance retention after 1000 and 5000 cycles respectively signifying the astonishing mechanical durability and long-term stability. All the properties exhibited by the fabricated electrode make it convenient for making flexible energy storage devices with the low form factor.Keywords: ZnO nanorods, electrospinning, mechanical endurance, flexible supercapacitor
Procedia PDF Downloads 28123817 Wound Healing Process Studied on DC Non-Homogeneous Electric Fields
Authors: Marisa Rio, Sharanya Bola, Richard H. W. Funk, Gerald Gerlach
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Cell migration, wound healing and regeneration are some of the physiological phenomena in which electric fields (EFs) have proven to have an important function. Physiologically, cells experience electrical signals in the form of transmembrane potentials, ion fluxes through protein channels as well as electric fields at their surface. As soon as a wound is created, the disruption of the epithelial layers generates an electric field of ca. 40-200 mV/mm, directing cell migration towards the wound site, starting the healing process. In vitro electrotaxis, experiments have shown cells respond to DC EFs polarizing and migrating towards one of the poles (cathode or anode). A standard electrotaxis experiment consists of an electrotaxis chamber where cells are cultured, a DC power source and agar salt bridges that help delaying toxic products from the electrodes to attain the cell surface. The electric field strengths used in such an experiment are uniform and homogeneous. In contrast, the endogenous electric field strength around a wound tend to be multi-field and non-homogeneous. In this study, we present a custom device that enables electrotaxis experiments in non-homogeneous DC electric fields. Its main feature involves the replacement of conventional metallic electrodes, separated from the electrotaxis channel by agarose gel bridges, through electrolyte-filled microchannels. The connection to the DC source is made by Ag/AgCl electrodes, incased in agarose gel and placed at the end of each microfluidic channel. An SU-8 membrane closes the fluidic channels and simultaneously serves as the single connection from each of them to the central electrotaxis chamber. The electric field distribution and current density were numerically simulated with the steady-state electric conduction module from ANSYS 16.0. Simulation data confirms the application of nonhomogeneous EF of physiological strength. To validate the biocompatibility of the device cellular viability of the photoreceptor-derived 661W cell line was accessed. The cells have not shown any signs of apoptosis, damage or detachment during stimulation. Furthermore, immunofluorescence staining, namely by vinculin and actin labelling, allowed the assessment of adhesion efficiency and orientation of the cytoskeleton, respectively. Cellular motility in the presence and absence of applied DC EFs was verified. The movement of individual cells was tracked for the duration of the experiments, confirming the EF-induced, cathodal-directed motility of the studied cell line. The in vitro monolayer wound assay, or “scratch assay” is a standard protocol to quantitatively access cell migration in vitro. It encompasses the growth of a confluent cell monolayer followed by the mechanic creation of a scratch, representing a wound. Hence, wound dynamics was monitored over time and compared for control and applied the electric field to quantify cellular population motility.Keywords: DC non-homogeneous electric fields, electrotaxis, microfluidic biochip, wound healing
Procedia PDF Downloads 27023816 UV-Reactive Electrospinning: Preparation, Characterization and Cell Culture Applications of Nanofiber Scaffolds Containing Keratin
Authors: Duygu Yüksel Deniz, Memet Vezir Kahraman, Serap Erdem Kuruca, Mediha Süleymanoğlu
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Our first aim was to synthesize Hydroxy Apatite (HAP) and then modify its surface by adding 4-Vinylbenzene boronic acid (4-VBBA). The characterization was done by FT-IR. By adding Polyvinyl alcohol (PVA) to 4- VBBA-HAP, we obtained a suitable electrospinning solution. PVA solution which was also modified by using alkoxy silanes, in order to prevent the scaffolds from being damaged by aqueous cell medium, was added. Keratin was dissolved and then added into the electrospinning solution. Keratin containing 4-VBBA- HAP/PVA composite was used to fabricate nanofiber scaffolds with the simultaneous UV-reactive electrospinning technique. The structural characterization was done by FT-IR. Thermal gravimetric analysis was also performed by using TGA. The morphological characterization was determined by SEM analyses. Our second aim was to create a scaffold where cells could grow. With this purpose, suitable nanofibers were choosen according to their SEM analysis. Keratin containing nanofibers were seeded with 3T3, ECV and SAOS cells and their cytotoxicity and cell proliferation were investigated by using MTT assay. After cell culturing process morphological characterization was determined by SEM analyses. These scaffolds were designed to be nontoxic biomaterials. Here, a comparision was made between keratin containing 3T3, ECV and SAOS seeded nanofiber scaffolds and the results were presented and discussed.Keywords: cell culture, keratin, nanofibers, UV-reactive electrospinning
Procedia PDF Downloads 45423815 A Prenylflavanoid, HME5 with Antiproliferative Activity in Human Ovarian Cancer Cells
Authors: Mashitoh Abd Rahman, Najihah Mohd Hashim, Faiqah Ramli, Syam Mohan, Noraziah Nordin, Hamed Karimian, Hapipah Mohd Ali
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Ovarian cancer is the most lethal gynecological malignancies. HME5, a prenylflavanoid has been isolated from local medicinal plant. This compound has been reported to possess a broad spectrum of biological activities including anticancer property. However, the potential of HME5 as an antiproliferative and cytotoxic agent on an ovarian cancer cells has not yet been investigated. In this present study, we examined the antiproliferative and cytotoxic effect of HME5 on Caov-3 (Human Ovarian Adenocarcinoma) cell line by using 3-[4,5-dimethylthizol-2-y]-2,5-diphenyltetrazolium bromide (MTT) assay, Acridine orange and propidium Iodide (AOPi) and cell cycle analysis study. HME5 has shown to inhibit Caov-3 in a time-dependent manner with the IC50 values of 5µg/ml, 2µg/ml and 1µg/ml after 24h, 48h and 72h treatment, respectively. Morphological study from AOPi analysis showed that HME5 induced apoptosis after 24 and 48h post-treatment. Nevertheless, HME5 exhibited cell cycle arrest at G1 phase as indicated in flow cytometry cell cycle profiling. In conclusion, HME5 inhibited proliferation of Caov-3 through induction of apoptosis and cell cycle arrest at G1 phase.Keywords: apoptosis, prenylflavanoid, ovarian cancer, HME5
Procedia PDF Downloads 46123814 Effect of Methanol Root Extracts of Moringa Oleifera on Lipid Profile Parameters, Atherogenic Indices and HMG – CoA Reductase Activities of Poloxamer 407-Induced Hyperlipidemic Rats
Authors: Matthew Ocheleka Itodo, Ogo Agbo Ogo, Agnes Ogbene Abutu, Bawa Inalegwu
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Hyperlipidemia is characterised by elevated serum total cholesterol and low density and very low-density lipoprotein cholesterol and decreased high-density lipoprotein are the risk factor for coronary heart diseases. There are claims by traditional medicine practitioners in Nigeria that Moringa oleifera plants are used for the treatment of cardiovascular diseases, but it appears there is no scientific research and, publication or documented work to verify these claims. This study aimed to determine the effect of methanol root extracts of Moringa oleifera on Lipid profile, Atherogenic indices and 3 hydroxyl 3 methylglutaryl Coenzyme A reductase activity of poloxamer 407-induced hyperlipidemic rats. The animals were grouped into 8; Group 1: Normal control, Group 2: Hyperlipidemic control. Groups 2 to 8 were induced with Poloxamer 407 1000 mg/Kg body weight. However, group 3 were treated with standard drugs (atorvastatin). Group 4 was treated with crude extract, and groups 5 to 8 were treated with purified fractions from column chromatography. The preliminary antihyperlipidemic study showed Methanol root extract at 200 mg/kg body weight significantly (p≤0.05) decreased total cholesterol, low-density lipoprotein, triacylglyceride, 3 hydroxyls 3 methylglutaryl Coenzyme A reductase, and increase high-density lipoprotein of hyperlipidemic treated groups. Screening the extracts for the most potent anti-hyperlipidemic activity reveals that fraction 1 of Total Cholesterol and Fraction 3 of Triacylglyceride have the highest percentage reduction of 56% and 51%, respectively. The atherogenic risk factor of all induced treated rats shows a significant (p<0.05) decrease in levels of Castelli’s risk index II, atherogenic index of plasma and a significant (p<0.05) higher level of Castelli’s risk index I ratio. The study shows that the methanol extract of root possesses antihyperlipidemic effects and may explain why it has been found to be useful in the management of cardiovascular diseases by traditional medicine practitioners.Keywords: hyperlipidemia, moringa oleifera, poloxamer 407, lipid profile
Procedia PDF Downloads 9023813 Track Initiation Method Based on Multi-Algorithm Fusion Learning of 1DCNN And Bi-LSTM
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Aiming at the problem of high-density clutter and interference affecting radar detection target track initiation in ECM and complex radar mission, the traditional radar target track initiation method has been difficult to adapt. To this end, we propose a multi-algorithm fusion learning track initiation algorithm, which transforms the track initiation problem into a true-false track discrimination problem, and designs an algorithm based on 1DCNN(One-Dimensional CNN)combined with Bi-LSTM (Bi-Directional Long Short-Term Memory )for fusion classification. The experimental dataset consists of real trajectories obtained from a certain type of three-coordinate radar measurements, and the experiments are compared with traditional trajectory initiation methods such as rule-based method, logical-based method and Hough-transform-based method. The simulation results show that the overall performance of the multi-algorithm fusion learning track initiation algorithm is significantly better than that of the traditional method, and the real track initiation rate can be effectively improved under high clutter density with the average initiation time similar to the logical method.Keywords: track initiation, multi-algorithm fusion, 1DCNN, Bi-LSTM
Procedia PDF Downloads 9523812 Molecular Motors in Smart Drug Delivery Systems
Authors: Ainoa Guinart, Maria Korpidou, Daniel Doellerer, Cornelia Palivan, Ben L. Feringa
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Stimuli responsive systems arise from the need to meet unsolved needs of current molecular drugs. Our study presents the design of a delivery system with high spatiotemporal control and tuneable release profiles. We study the incorporation of a hydrophobic synthetic molecular motor into PDMS-b-PMOXA block copolymer vesicles to create a self-assembled system. We prove their successful incorporation and selective activation by low powered visible light (λ 430 nm, 6.9 mW). We trigger the release of a fluorescent dye with high release efficiencies over sequential cycles (up to 75%) with the ability to turn on and off the release behaviour on demand by light irradiation. Low concentrations of photo-responsive units are proven to trigger release down to 1 mol% of molecular motor. Finally, we test our system in relevant physiological conditions using a lung cancer cell line and the encapsulation of an approved drug. Similar levels of cell viability are observed compared to the free-given drugshowing the potential of our platform to deliver functional drugs on demand with the same efficiency and lower toxicity.Keywords: molecular motor, polymer, drug delivery, light-responsive, cancer, selfassembly
Procedia PDF Downloads 13523811 Plasma Lipid Profiles and Atherogenic Indices of Rats Fed Raw and Processed Jack Fruit (Artocarpus heterophyllus) Seeds Diets at Different Concentrations
Authors: O. E. Okafor, L. U. S. Ezeanyika, C. G. Nkwonta, C. J. Okonkwo
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The effect of processing on plasma lipid profile and atherogenic indices of rats fed Artocarpus heterophyllus seed diets at different concentrations were investigated. Fifty five rats were used for this study, they were divided into eleven groups of five rats each (one control group and ten test groups), the test groups were fed raw, boiled, roasted, fermented, and soaked diets at 10 % and 40% concentrations. The study lasted for thirty five days. The diets led to significant decrease (p < 0.05) in plasma cholesterol and triacylglycerol of rats fed 10% and 40% concentrations of the diets, and a significant increase (p < 0.05) in high density lipoprotein (HDL) levels at 40% concentrations of the test diets. The diets also produced decrease in low density lipoprotein (LDL), very low density lipoprotein (VLDL), cardiac risk ratio (CRR), atherogenic index of plasma (AIP) and atherogenic coefficient (AC) at 40% concentrations except the soaked group that showed slight elevation of LDL, CRR, AC and AIP at 40% concentration. Artocarpus heterophyllus seeds could be beneficial to health because of its ability to increase plasma HDL and reduce plasma LDL, VLDL, cholesterol, triglycerides and atherogenic indices at higher diet concentration.Keywords: artocarpus heterophyllus, atherogenic indices, concentrations, lipid profile
Procedia PDF Downloads 30223810 New Quinazoline Derivative Exhibit Cytotoxic Effect agaisnt MCF-7 Human Breast Cancer Cell
Authors: Maryam Zahedifard, Fadhil Lafta Faraj, Nazia Abdul Majid, Hapipah Mohd Ali, Mahmood Ameen Abdulla
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The new quinazoline Schiff bases have been synthesized through condensation reaction of 2-aminobenzhydrazide with 5-bromosalicylaldehyde and 3-methoxy-5-bromosalicylaldehyde. The compound was investigated for anticancer activity against MCF-7 human breast cancer cell line. It demonstrated a remarkable antiproliferative effect, with an IC50 value of 3.41±0.34, after 72 hours of treatment. Most apoptosis morphological features in treated MCF-7 cells were observed by AO/PI staining. The results of cell cycle analysis indicate that compounds did not induce S and M phase arrest in cell after 24 hours of treatment. Furthermore, MCF-7 cells treated with compound subjected to apoptosis death, as exhibited by perturbation of mitochondrial membrane potential and cytochrome C release as well as increase in ROS generation. We also found activation of caspases 3/7 and -9. Moreover, acute toxicity results demonstrated the nontoxic nature of the compounds in mice. Our results showed the selected compound significantly induce apoptosis in MCF-7 cells via intrinsic pathway, which might be considered as a potential candidate for further in vivo and clinical breast cancer studies.Keywords: quinazoline Schiff base, apoptosis, MCF-7, caspase, cell cycle, acute toxicity
Procedia PDF Downloads 44223809 Investigation and Monitoring Method of Vector Density in Kaohsiung City
Authors: Chiu-Wen Chang, I-Yun Chang, Wei-Ting Chen, Hui-Ping Ho, Chao-Ying Pan, Joh-Jong Huang
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Dengue is a ‘community disease’ or ‘environmental disease’, as long as the environment exist suitable container (including natural and artificial) for mosquito breeding, once the virus invade will lead to the dengue epidemic. Surveillance of vector density is critical to effective infectious disease control and play an important role in monitoring the dynamics of mosquitoes in community, such as mosquito species, density, distribution area. The objective of this study was to examine the relationship in vector density survey (Breteau index, Adult index, House index, Container index, and Larvae index) form 2014 to 2016 in Kaohsiung City and evaluate the effects of introducing the Breeding Elimination and Appraisal Team (hereinafter referred to as BEAT) as an intervention measure on eliminating dengue vector breeding site started from May 2016. BEAT were performed on people who were suspected of contracting dengue fever, a surrounding area measuring 50 meters by 50 meters was demarcated as the emergency prevention and treatment zone. BEAT would perform weekly vector mosquito inspections and vector mosquito inspections in regions with a high Gravitrap index and assign a risk assessment index to each region. These indices as well as the prevention and treatment results were immediately reported to epidemic prevention-related units every week. The results indicated that, vector indices from 2014 to 2016 showed no statistically significant differences in the Breteau index, adult index, and house index (p > 0.05) but statistically significant differences in the container index and larvae index (p <0.05). After executing the integrated elimination work, container index and larvae index are statistically significant different from 2014 to 2016 in the (p < 0.05). A post hoc test indicated that the container index of 2014 (M = 12.793) was significantly higher than that of 2016 (M = 7.631), and that the larvae index of 2015 (M = 34.065) was significantly lower than that of 2014 (M = 66.867). The results revealed that effective vector density surveillance could highlight the focus breeding site and then implement the immediate control action (BEAT), which successfully decreased the vector density and the risk of dengue epidemic.Keywords: Breteau index, dengue control, monitoring method, vector density
Procedia PDF Downloads 19823808 The Role of P2X7 Cytoplasmic Anchor in Inflammation
Authors: Federico Cevoli
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Purinergic P2X7 receptors (P2X7R) are ligand-gated non-selective cation channels involved in several physiological and pathological processes. They are particularly promising pharmacological targets as they are present in an increasing number of different cells types. P2X7R activation is triggered following elevated concentrations of extracellular ATP, similarly to those observed in tissues injury, chronic inflammation and T-cell activation, as well as in the scrambling of phospholipids leading to membrane blebbing and apoptosis. Another hallmark of P2X7 is cell permeabilization, commonly known as “macropore” formation allowing the passage of nanometer-sized molecules up to 900Da. Recently, full-length P2X7 Cryo-EM structures revealed unique functional sites, including two cytoplasmic domains - the cytoplasmic "anchor" and "ballast". To date, the molecular units/complex by which P2X7R exerts its pathophysiological functions are unknown. Using custom-made cell-penetrating HIV-1 TAT peptides, we show for the first-time potential implications of P2X7 cytoplasmic anchor in the regulation of caspase3/7 activation as well as TNFα regulation.Keywords: P2X7R, immunology, TAT-peptide, cell death
Procedia PDF Downloads 13723807 Surface Adjustments for Endothelialization of Decellularized Porcine Pericardium
Authors: M. Markova, E. Filova, O. Kaplan, R. Matejka, L. Bacakova
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The porcine pericardium is used as a material for cardiac and aortic valves substitutes. Current biological aortic heart valve prosthesis have a limited lifetime period because they undergo degeneration. In order to make them more biocompatible and prolong their lifetime it is necessary to reseed the decellularized prostheses with endothelial cells and with valve interstitial cells. The endothelialization of the prosthesis-surface may be supported by suitable chemical surface modification of the prosthesis. The aim of this study is to prepare bioactive fibrin layers which would both support endothelialization of porcine pericardium and enhance differentiation and maturation of the endothelial cells seeded. As a material for surface adjustments we used layers of fibrin with/without heparin and some of them with adsorbed or chemically bound FGF2, VEGF or their combination. Fibrin assemblies were prepared in 24-well cell culture plate and were seeded with HSVEC (Human Saphenous Vein Endothelial Cells) at a density of 20,000 cells per well in EGM-2 medium with 0.5% FS and without heparin, without FGF2 and without VEGF; medium was supplemented with aprotinin (200 U/mL). As a control, surface polystyrene (PS) was used. Fibrin was also used as homogeneous impregnation of the decellularized porcine pericardium throughout the scaffolds. Morphology, density, and viability of the seeded endothelial cells were observed from micrographs after staining the samples by LIVE/DEAD cytotoxicity/viability assay kit on the days 1, 3, and 7. Endothelial cells were immunocytochemically stained for proteins involved in cell adhesion, i.e. alphaV integrin, vinculin, and VE-cadherin, markers of endothelial cells differentiation and maturation, i.e. von Willebrand factor and CD31, and for extracellular matrix proteins typically produced by endothelial cells, i.e. type IV collagen and laminin. The staining intensities were subsequently quantified using a software. HSVEC cells grew on each of the prepared surfaces better than on control surface. They reached confluency. The highest cell densities were obtained on the surface of fibrin with heparin and both grow factors used together. Intensity of alphaV integrins staining was highest on samples with remained fibrin layer, i.e. on layers with lower cell densities, i.e. on fibrin without heparin. Vinculin staining was apparent, but was rather diffuse, on fibrin with both FGF2 and VEGF and on control PS. Endothelial cells on all samples were positively stained for von Willebrand factor and CD31. VE-cadherin receptors clusters were best developed on fibrin with heparin and growth factors. Significantly stronger staining of type IV collagen was observed on fibrin with heparin and both growth factors. Endothelial cells on all samples produced laminin-1. Decellularized pericardium was homogeneously filled with fibrin structures. These fibrin-modified pericardium samples will be further seeded with cells and cultured in a bioreactor. Fibrin layers with/without heparin and with adsorbed or chemically bound FGF2, VEGF or their combination are good surfaces for endothelialization of cardiovascular prostheses or porcine pericardium based heart valves. Supported by the Ministry of Health, grants No15-29153A and 15-32497A, and the Grant Agency of the Czech Republic, project No. P108/12/G108.Keywords: aortic valves prosthesis, FGF2, heparin, HSVEC cells, VEGF
Procedia PDF Downloads 26523806 Flame Spray Pyrolysis as a High-Throughput Method to Generate Gadolinium Doped Titania Nanoparticles for Augmented Radiotherapy
Authors: Malgorzata J. Rybak-Smith, Benedicte Thiebaut, Simon Johnson, Peter Bishop, Helen E. Townley
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Gadolinium doped titania (TiO2:Gd) nanoparticles (NPs) can be activated by X-ray radiation to generate Reactive Oxygen Species (ROS), which can be effective in killing cancer cells. As such, treatment with these NPs can be used to enhance the efficacy of conventional radiotherapy. Incorporation of the NPs in to tumour tissue will permit the extension of radiotherapy to currently untreatable tumours deep within the body, and also reduce damage to neighbouring healthy cells. In an attempt to find a fast and scalable method for the synthesis of the TiO2:Gd NPs, the use of Flame Spray Pyrolysis (FSP) was investigated. A series of TiO2 NPs were generated with 1, 2, 5 and 7 mol% gadolinium dopant. Post-synthesis, the TiO2:Gd NPs were silica-coated to improve their biocompatibility. Physico-chemical characterisation was used to determine the size and stability in aqueous suspensions of the NPs. All analysed TiO2:Gd NPs were shown to have relatively high photocatalytic activity. Furthermore, the FSP synthesized silica-coated TiO2:Gd NPs generated enhanced ROS in chemico. Studies on rhabdomyosarcoma (RMS) cell lines (RD & RH30) demonstrated that in the absence of irradiation all TiO2:Gd NPs were inert. However, application of TiO2:Gd NPs to RMS cells, followed by irradiation, showed a significant decrease in cell proliferation. Consequently, our studies showed that the X-ray-activatable TiO2:Gd NPs can be prepared by a high-throughput scalable technique to provide a novel and affordable anticancer therapy.Keywords: cancer, gadolinium, ROS, titania nanoparticles, X-ray
Procedia PDF Downloads 43123805 Study of the Combinatorial Impact of Substrate Properties on Mesenchymal Stem Cell Migration Using Microfluidics
Authors: Nishanth Venugopal Menon, Chuah Yon Jin, Samantha Phey, Wu Yingnan, Zhang Ying, Vincent Chan, Kang Yuejun
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Cell Migration is a vital phenomenon that the cells undergo in various physiological processes like wound healing, disease progression, embryogenesis, etc. Cell migration depends primarily on the chemical and physical cues available in the cellular environment. The chemical cue involves the chemokines secreted and gradients generated in the environment while physical cues indicate the impact of matrix properties like nanotopography and stiffness on the cells. Mesenchymal Stem Cells (MSCs) have been shown to have a role wound healing in vivo and its migration to the site of the wound has been shown to have a therapeutic effect. In the field of stem cell based tissue regeneration of bones and cartilage, one approach has been to introduce scaffold laden with MSCs into the site of injury to enable tissue regeneration. In this work, we have studied the combinatorial impact of the substrate physical properties on MSC migration. A microfluidic in vitro model was created to perform the migration studies. The microfluidic model used is a three compartment device consisting of two cell seeding compartments and one migration compartment. Four different PDMS substrates with varying substrate roughness, stiffness and hydrophobicity were created. Its surface roughness and stiffness was measured using Atomic Force Microscopy (AFM) while its hydrphobicity was measured from the water contact angle using an optical tensiometer. These PDMS substrates are sealed to the microfluidic chip following which the MSCs are seeded and the cell migration is studied over the period of a week. Cell migration was quantified using fluorescence imaging of the cytoskeleton (F-actin) to find out the area covered by the cells inside the migration compartment. The impact of adhesion proteins on cell migration was also quantified using a real-time polymerase chain reaction (qRT PCR). These results suggested that the optimal substrate for cell migration would be one with an intermediate level of roughness, stiffness and hydrophobicity. A higher or lower value of these properties affected cell migration negatively. These observations have helped us in understanding that different substrate properties need to be considered in tandem, especially while designing scaffolds for tissue regeneration as cell migration is normally impacted by the combinatorial impact of the matrix. These observations may lead us to scaffold optimization in future tissue regeneration applications.Keywords: cell migration, microfluidics, in vitro model, stem cell migration, scaffold, substrate properties
Procedia PDF Downloads 55723804 Enhanced Field Emission from Plasma Treated Graphene and 2D Layered Hybrids
Authors: R. Khare, R. V. Gelamo, M. A. More, D. J. Late, Chandra Sekhar Rout
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Graphene emerges out as a promising material for various applications ranging from complementary integrated circuits to optically transparent electrode for displays and sensors. The excellent conductivity and atomic sharp edges of unique two-dimensional structure makes graphene a propitious field emitter. Graphene analogues of other 2D layered materials have emerged in material science and nanotechnology due to the enriched physics and novel enhanced properties they present. There are several advantages of using 2D nanomaterials in field emission based devices, including a thickness of only a few atomic layers, high aspect ratio (the ratio of lateral size to sheet thickness), excellent electrical properties, extraordinary mechanical strength and ease of synthesis. Furthermore, the presence of edges can enhance the tunneling probability for the electrons in layered nanomaterials similar to that seen in nanotubes. Here we report electron emission properties of multilayer graphene and effect of plasma (CO2, O2, Ar and N2) treatment. The plasma treated multilayer graphene shows an enhanced field emission behavior with a low turn on field of 0.18 V/μm and high emission current density of 1.89 mA/cm2 at an applied field of 0.35 V/μm. Further, we report the field emission studies of layered WS2/RGO and SnS2/RGO composites. The turn on field required to draw a field emission current density of 1μA/cm2 is found to be 3.5, 2.3 and 2 V/μm for WS2, RGO and the WS2/RGO composite respectively. The enhanced field emission behavior observed for the WS2/RGO nanocomposite is attributed to a high field enhancement factor of 2978, which is associated with the surface protrusions of the single-to-few layer thick sheets of the nanocomposite. The highest current density of ~800 µA/cm2 is drawn at an applied field of 4.1 V/μm from a few layers of the WS2/RGO nanocomposite. Furthermore, first-principles density functional calculations suggest that the enhanced field emission may also be due to an overlap of the electronic structures of WS2 and RGO, where graphene-like states are dumped in the region of the WS2 fundamental gap. Similarly, the turn on field required to draw an emission current density of 1µA/cm2 is significantly low (almost half the value) for the SnS2/RGO nanocomposite (2.65 V/µm) compared to pristine SnS2 (4.8 V/µm) nanosheets. The field enhancement factor β (~3200 for SnS2 and ~3700 for SnS2/RGO composite) was calculated from Fowler-Nordheim (FN) plots and indicates emission from the nanometric geometry of the emitter. The field emission current versus time plot shows overall good emission stability for the SnS2/RGO emitter. The DFT calculations reveal that the enhanced field emission properties of SnS2/RGO composites are because of a substantial lowering of work function of SnS2 when supported by graphene, which is in response to p-type doping of the graphene substrate. Graphene and 2D analogue materials emerge as a potential candidate for future field emission applications.Keywords: graphene, layered material, field emission, plasma, doping
Procedia PDF Downloads 36123803 Combining Transcriptomics, Bioinformatics, Biosynthesis Networks and Chromatographic Analyses for Cotton Gossypium hirsutum L. Defense Volatiles Study
Authors: Ronald Villamar-Torres, Michael Staudt, Christopher Viot
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Cotton Gossypium hirsutum L. is one of the most important industrial crops, producing the world leading natural textile fiber, but is very prone to arthropod attacks that reduce crop yield and quality. Cotton cultivation, therefore, makes an outstanding use of chemical pesticides. In reaction to herbivorous arthropods, cotton plants nevertheless show natural defense reactions, in particular through volatile organic compounds (VOCs) emissions. These natural defense mechanisms are nowadays underutilized but have a very high potential for cotton cultivation, and elucidating their genetic bases will help to improve their use. Simulating herbivory attacks by mechanical wounding of cotton plants in greenhouse, we studied by qPCR the changes in gene expression for genes of the terpenoids biosynthesis pathway. Differentially expressed genes corresponded to higher levels of the terpenoids biosynthesis pathway and not to enzymes synthesizing particular terpenoids. The genes were mapped on the G. hirsutum L. reference genome; their global relationships inside the general metabolic pathways and the biosynthesis of secondary metabolites were visualized with iPath2. The chromatographic profiles of VOCs emissions indicated first monoterpenes and sesquiterpenes emissions, dominantly four molecules known to be involved in plant reactions to arthropod attacks. As a result, the study permitted to identify potential key genes for the emission of volatile terpenoids by cotton plants in reaction to an arthropod attack, opening possibilities for molecular-assisted cotton breeding in benefit of smallholder cotton growers.Keywords: biosynthesis pathways, cotton, mechanisms of plant defense, terpenoids, volatile organic compounds
Procedia PDF Downloads 37423802 Effects of Coupling Agent on the Properties of Henequen Microfiber (NF) Filled High Density Polyethylene (HDPE) Composites
Authors: Pravin Gaikwad, Prakash Mahanwar
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The main objective of incorporating natural fibers such as Henequen microfibers (NF) into the High-Density Polyethylene (HDPE) polymer matrix is to reduce the cost and to enhance the mechanical as well as other properties. The Henequen microfibers were chopped manually to 5-7mm in length and added into the polymer matrix at the optimized concentration of 8 wt %. In order to facilitate the link between Henequen microfibers (NF) and HDPE matrix, coupling agent such as Glycidoxy (Epoxy) Functional Methoxy Silane (GPTS) at various concentrations from 0.1%, 0.3%, 0.5%, 0.7%, 0.9%, and 1% by weight to the total fibers were added. The tensile strength of the composite increased marginally while % elongation at break of the composites decreased with increase in silane loading by wt %. Tensile modulus and stiffness observed increased at 0.9 wt % GPTS loading. Flexural as well as impact strength of the composite decreased with increase in GPTS loading by weight %. Dielectric strength of the composite also found increased marginally upto 0.5wt % silane loading and thereafter remained constant.Keywords: Henequen microfibers (NF), polymer composites, HDPE, coupling agent, GPTS
Procedia PDF Downloads 43923801 Improving Radiation Efficiency Using Metamaterial in Pyramidal Horn Antenna
Authors: Amit Kumar Baghel, Sisir Kumar Nayak
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The proposed metamaterial design help to increase the radiation efficiency at 2.9 GHz by reducing the side and back lobes by making the phase difference of the waves emerging from the phase center of the horn antenna same after passing through metamaterial array. The unit cell of the metamaterial is having concentric ring structure made of copper of 0.035 mm thickness on both sides of FR4 sheet. The inner ring diameter is kept as 3 mm, and the outer ring diameters are changed according to the path and tramission phase difference of the unit cell from the phase center of the antenna in both the horizontal and vertical direction, i.e., in x- and y-axis. In this case, the ring radius varies from 3.19 mm to 6.99 mm with the respective S21 phase difference of -62.25° to -124.64°. The total phase difference can be calculated by adding the path difference of the respective unit cell in the array to the phase difference of S21. Taking one of the unit cell as the reference, the total phase difference between the reference unit cell and other cells must be integer multiple of 360°. The variation of transmission coefficient S21 with the ring radius is greater than -6 dB. The array having 5 x 5 unit cell is kept inside the pyramidal horn antenna (L X B X H = 295.451 x 384.233 x 298.66 mm3) at a distance of 36.68 mm from the waveguide throat. There is an improvement in side lobe level in E-plane by 14.6 dB when the array is used. The front to back lobe ration is increased by 1 dB by using the array. The proposed antenna with metamaterial array can be used in beam shaping for wireless power transfer applications.Keywords: metamaterial, side lobe level, front to back ratio, beam forming
Procedia PDF Downloads 27623800 Localising Gauss’s Law and the Electric Charge Induction on a Conducting Sphere
Authors: Sirapat Lookrak, Anol Paisal
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Space debris has numerous manifestations, including ferro-metalize and non-ferrous. The electric field will induce negative charges to split from positive charges inside the space debris. In this research, we focus only on conducting materials. The assumption is that the electric charge density of a conducting surface is proportional to the electric field on that surface due to Gauss's Law. We are trying to find the induced charge density from an external electric field perpendicular to a conducting spherical surface. An object is a sphere on which the external electric field is not uniform. The electric field is, therefore, considered locally. The localised spherical surface is a tangent plane, so the Gaussian surface is a very small cylinder, and every point on a spherical surface has its own cylinder. The electric field from a circular electrode has been calculated in near-field and far-field approximation and shown Explanation Touchless maneuvering space debris orbit properties. The electric charge density calculation from a near-field and far-field approximation is done.Keywords: near-field approximation, far-field approximation, localized Gauss’s law, electric charge density
Procedia PDF Downloads 13223799 Expression of Human Papillomavirus Type 18 L1 Virus-Like Particles in Methylotropic Yeast, Pichia Pastoris
Authors: Hossein Rassi, Marjan Moradi Fard, Samaneh Niko
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Human papillomavirus type 16 and 18 are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, HPV type 18 accounts for about 34 % of all HPV infections in Iran and the most promising vaccine against HPV infection is based on the L1 major capsid protein. The L1 protein of HPV18 has the capacity to self-assemble into capsomers or virus-like particles (VLPs) that are non-infectious, highly immunogenic and allowing their use in vaccine production. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system used to produce high levels of heterologous proteins. In this study we expressed HPV18 L1 VLPs in P. pastoris. The gene encoding the major capsid protein L1 of the high-risk HPV type 18 was isolated from Iranian patient by PCR and inserted into pTG19-T vector to obtain the recombinant expression vector pTG19-HPV18-L1. Then, the pTG19-HPV18-L1 was transformed into E. coli strain DH5α and the recombinant protein HPV18 L1 was expressed under IPTG induction in soluble form. The HPV18 L1 gene was excised from recombinant plasmid with XhoI and EcoRI enzymes and ligated into the yeast expression vector pPICZα linearized with the same enzymes, and transformed into P. pastoris. Induction and expression of HPV18 L1 protein was demonstrated by BMGY/BMMY and RT PCR. The parameters for induced cultivation for strain in P. pastoris KM71 with HPV16L1 were investigated in shaking flask cultures. After induced cultivation BMMY (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 24 h at 37 degrees C for 96 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by P. pastoris for HPV-18 L1 gene. The VLP-based HPV vaccines can prevent persistent HPV18 infections and cervical cancer in Iran. The HPV-18 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV-18 L1 vaccine in human. Also, HPV type 6 L1 proteins expressed in Pichia pastoris will facilitate the HPV vaccine development and structure-function study.Keywords: Pichia pastoris, L1 virus-like particles, human papillomavirus type 18, biotechnology
Procedia PDF Downloads 40723798 Growth of Algal Biomass in Laboratory and in Pilot-Scale Algal Photobioreactors in the Temperate Climate of Southern Ireland
Authors: Linda A. O’Higgins, Astrid Wingler, Jorge Oliveira
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The growth of Chlorella vulgaris was characterized as a function of irradiance in a laboratory turbidostat (1 L) and compared to batch growth in sunlit modules (5–25 L) of the commercial Phytobag photobioreactor. The effects of variable sunlight and culture density were deconvoluted by a mathematical model. The analysis showed that algal growth was light-limited due to shading by external construction elements and due to light attenuation within the algal bags. The model was also used to predict maximum biomass productivity. The manipulative experiments and the model predictions were confronted with data from a production season of a 10m2 pilot-scale photobioreactor, Phytobag (10,000 L). The analysis confirmed light limitation in all three photobioreactors. An additional limitation of biomass productivity was caused by the nitrogen starvation that was used to induce lipid accumulation. Reduction of shading and separation of biomass and lipid production are proposed for future optimization.Keywords: microalgae, batch cultivation, Chlorella vulgaris, Mathematical model, photobioreactor, scale-up
Procedia PDF Downloads 11623797 Phenotype of Cutaneous Squamous Cell Carcinoma in a Brazilian City with a Tropical Climate
Authors: Julia V. F. Cortes, Maria E. V. Amarante, Carolina L. Cerdeira, Roberta B. V. Silva
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Nonmelanoma skin cancer is more commonly diagnosed than all other malignancies combined. In that group, cutaneous squamous cell carcinoma stands out for having the highest probability of metastasis and recurrence after treatment, in addition to being the second most prevalent form of skin cancer. Its main risk factors include exposure to carcinogens, such as ultraviolet radiation related to sunlight exposure, smoking, alcohol consumption, and human papillomavirus (HPV) infection. Considering the increased risk of skin cancer in the Brazilian population, caused by the high incidence of solar radiation, and the importance of identifying risk phenotypes for the accomplishment of public health actions, an epidemiological study was conducted in a city with a tropical climate located in southeastern Brazil, aiming to identify the target population and assist in primary and secondary prevention. This study describes the profile of patients with cutaneous squamous cell cancer, correlating the variables, sex, age, and differentiation. The study used as primary data source the results of anatomopathological exams delivered from January 2015 to December 2019 for patients registered at one pathology service, which analyzes the results of biopsies, Thus, 66 patients with cutaneous squamous cell carcinoma were analyzed. The most affected age group was 60 years or older (78.79%), emphasizing that moderately differentiated (79.49%) and well-differentiated forms (66.67%) are prevalent in this age group, resulting in a difference of 12.82 percentage points between them. In addition, the predominant sex was male (58%), and it was found that half of the women and 65.79% of men had a moderately differentiated type, whereas the well-differentiated type was slightly more frequent in women. It is worth noting that the moderately differentiated subtype has a 59.20% prevalence among all cases. Thus, it was concluded that the most affected age group was 60 years or older and that men were more affected. As for the subtype, the moderately differentiated one, which is recognized for presenting the second-highest risk for metastasis, was prevalent in this study, affecting 6.6% more men and predominating in the elderly.Keywords: cutaneous squamous cell carcinoma, epidemiology, skin cancer, spinal cell cancer
Procedia PDF Downloads 11723796 An Epidemiological Study on Cutaneous Melanoma, Basocellular and Epidermoid Carcinomas Diagnosed in a Sunny City in Southeast Brazil in a Five-Year Period
Authors: Carolina L. Cerdeira, Julia V. F. Cortes, Maria E. V. Amarante, Gersika B. Santos
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Skin cancer is the most common cancer in several parts of the world; in a tropical country like Brazil, the situation isn’t different. The Brazilian population is exposed to high levels of solar radiation, increasing the risk of developing cutaneous carcinoma. Aimed at encouraging prevention measures and the early diagnosis of these tumors, a study was carried out that analyzed data on cutaneous melanomas, basal cell, and epidermoid carcinomas, using as primary data source the medical records of 161 patients registered in one pathology service, which performs skin biopsies in a city of Minas Gerais, Brazil. All patients diagnosed with skin cancer at this service from January 2015 to December 2019 were included. The incidence of skin carcinoma cases was correlated with the identification of histological type, sex, age group, and topographic location. Correlation between variables was verified by Fisher's exact test at a nominal significance level of 5%, with statistical analysis performed by R® software. A significant association was observed between age group and type of cancer (p=0.0085); age group and sex (0.0298); and type of cancer and body region affected (p < 0.01). Those 161 cases analyzed comprised 93 basal cell carcinomas, 66 epidermoid carcinomas, and only two cutaneous melanomas. In the group aged 19 to 30 years, the epidermoid form was most prevalent; from 31 to 45 and from 46 to 59 years, the basal cell prevailed; in 60-year-olds or over, both types had higher frequencies. Associating age group and sex, in groups aged 18 to 30 and 46 to 59 years, women were most affected. In the 31-to 45-year-old group, men predominated. There was a gender balance in the age group 60-year-olds or over. As for topography, there was a high prevalence in the head and neck, followed by upper limbs. Relating histological type and topography, there was a prevalence of basal cell and epidermoid carcinomas in the head and neck. In the chest, the basal cell form was most prevalent; in upper limbs, the epidermoid form prevailed. Cutaneous melanoma affected only the chest and upper limbs. About 82% of patients 60-year-olds or over had head and neck cancer; from 46 to 59 and 60-year-olds or over, the head and neck region and upper limbs were predominantly affected; the distribution was balanced in the 31-to 45-year-old group. In conclusion, basal cell carcinoma was predominant, whereas cutaneous melanoma was the rarest among the types analyzed. Patients 60-year-olds or over were most affected, showing gender balance. In young adults, there was a prevalence of the epidermoid form; in middle-aged patients, basal cell carcinoma was predominant; in the elderly, both forms presented with higher frequencies. There was a higher incidence of head and neck cancers, followed by malignancies affecting the upper limbs. The epidermoid type manifested significantly in the upper limbs. Body regions such as the thorax and lower limbs were less affected, which is justified by the lower exposure of these areas to incident solar radiation.Keywords: basal cell carcinoma, cutaneous melanoma, skin cancer, squamous cell carcinoma, topographic location
Procedia PDF Downloads 12923795 Patients Undergoing Diagnostic Bronchoscopy at General Ahmad Yani Hospital in 2023: A Profile of Lung Cancer Types
Authors: Devita Wardani, Pratiwi Gusti Wahyu, Andreas Infianto, Raden Dicky Wirawan Listiandoko, Descahirul, Yunnica
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Flexible fiberoptic bronchoscopy (FFB) is the essential method for both the diagnosis and staging of central lung cancer. Concerning central lesions, the sensitivity of endobronchial forceps biopsy of a visible endobronchial lesion is 74%. The aim is to determine the distribution of lung cancer types of patients diagnosed with and who underwent bronchoscopy at General Ahmad Yani Metro Hospital. Methods: This study is a cross-sectional descriptive study. Data are obtained from histopathological results of lung malignancy through bronchoscopy either from forceps biopsy, washing and brushing. Results: Lung cancer is more found in men (72.2%) than women (27.8%). The average age for men ranges 57 years old for Non-Small Cell and 56 years old for Small Cell case. Most histopathology found in non-small cell lung cancer (87.7%), with adenocarcinoma as the most common type (68.4%), followed by squamous cell carcinoma (29.1%). Findings for adenocarcinoma showed that men had an incidence of 64.8% and women had an incidence of 35.2%. Like NSCLC, men can have up to 90% of cases of SCLC compared to 10% in women. In addition to non-small cell and small cell types, other types, such as large cells and renal carcinoma metastases, were obtained. Conclusions: Adenocarcinoma lung cancer is the most widely obtained type with predominantly male patients.Keywords: bronchoscopy, forceps biopsy, washing, brushing
Procedia PDF Downloads 323794 Comparative Morphometric Analysis of Ambardi and Mangari Watersheds of Kadvi and Kasari River Sub-Basins in Kolhapur District, Maharashtra, India: Using Geographical Information System (GIS)
Authors: Chandrakant Gurav, Md. Babar
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In the present study, an attempt is made to delineate the comparative morphometric analysis of Ambardi and Mangari watersheds of Kadvi and Kasari rivers sub-basins, Kolhapur District, Maharashtra India, using Geographical Information System (GIS) techniques. GIS is a computer assisted information method to store, analyze and display spatial data. Both the watersheds originate from Masai plateau of Jotiba- Panhala Hill range in Panhala Taluka of Kolhapur district. Ambardi watersheds cover 42.31 Sq. km. area and occur in northern hill slope, whereas Mangari watershed covers 54.63 Sq. km. area and occur on southern hill slope. Geologically, the entire study area is covered by Deccan Basaltic Province (DBP) of late Cretaceous to early Eocene age. Laterites belonging to late Pleistocene age also occur in the top of the hills. The objective of the present study is to carry out the morphometric parameters of watersheds, which occurs in differing slopes of the hill. Morphometric analysis of Ambardi watershed indicates it is of 4th order stream and Mangari watershed is of 5th order stream. Average bifurcation ratio of both watersheds is 5.4 and 4.0 showing that in both the watersheds streams flow from homogeneous nature of lithology and there is no structural controlled in development of the watersheds. Drainage density of Ambardi and Mangari watersheds is 3.45 km/km2 and 3.81 km/km2 respectively, and Stream Frequency is 4.51 streams/ km2 and 5.97 streams/ km2, it indicates that high drainage density and high stream frequency is governed by steep slope and low infiltration rate of the area for groundwater recharge. Textural ratio of both the watersheds is 6.6 km-1 and 9.6 km-1, which indicates that the drainage texture is fine to very fine. Form factor, circularity ratio and elongation ratios of the Ambardi and Mangari watersheds shows that both the watersheds are elongated in shape. The basin relief of Ambardi watershed is 447 m, while Mangari is 456 m. Relief ratio of Ambardi is 0.0428 and Mangari is 0.040. The ruggedness number of Ambardi is 1.542 and Mangari watershed is 1.737. The ruggedness number of both the watersheds is high which indicates the relief and drainage density is high.Keywords: Ambardi, Deccan basalt, GIS, morphometry, Mangari, watershed
Procedia PDF Downloads 30123793 An Endophyte of Amphipterygium adstringens as Producer of Cytotoxic Compounds
Authors: Karol Rodriguez-Peña, Martha L. Macias-Rubalcava, Leticia Rocha-Zavaleta, Sergio Sanchez
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A bioassay-guided study for anti-cancer compounds from endophytes of the Mexican medicinal plant Amphipteryygium adstringens resulted in the isolation of a streptomycete capable of producing a group of compounds with high cytotoxic activity. Microorganisms from surface sterilized samples of various sections of the plant were isolated and all the actinomycetes found were evaluated for their potential to produce compounds with cytotoxic activity against cancer cell lines MCF7 (breast cancer) and HeLa (cervical cancer) as well as the non-tumoural cell line HaCaT (keratinocyte). The most active microorganism was picked for further evaluation. The identification of the microorganism was carried out by 16S rDNA gene sequencing, finding the closest proximity to Streptomyces scabrisporus, but with the additional characteristic that the strain isolated in this study was capable of producing colorful compounds never described for this species. Crude extracts of dichloromethane and ethyl acetate showed IC50 values of 0.29 and 0.96 μg/mL for MCF7, 0.51 and 1.98 μg/mL for HeLa and 0.96 and 2.7 μg/mL for HaCaT. Scaling the fermentation to 10 L in a bioreactor generated 1 g of total crude extract, which was fractionated by silica gel open column to yield 14 fractions. Nine of the fractions showed cytotoxic activity. Fraction 4 was chosen for subsequent purification because of its high activity against cancerous cell lines, lower activity against keratinocytes. HPLC-UV-MS/ESI was used for the evaluation of this fraction, finding at least 10 different compounds with high values of m/z (≈588). Purification of the compounds was carried out by preparative thin-layer chromatography. The prevalent compound was Steffimycin B, a molecule known for its antibiotic and cytotoxic activities and also for its low solubility in aqueous solutions. Along with steffimycin B, another five compounds belonging to the steffimycin family were isolated and at this moment their structures are being elucidated, some of which display better solubility in water: an attractive property for the pharmaceutical industry. As a conclusion to this study, the isolation of endophytes resulted in the discovery of a strain capable of producing compounds with high cytotoxic activity that need to be studied for their possible utilization.Keywords: amphipterygium adstringens, cytotoxicity, streptomyces scabrisporus, steffimycin
Procedia PDF Downloads 36423792 Automated Buffer Box Assembly Cell Concept for the Canadian Used Fuel Packing Plant
Authors: Dimitrie Marinceu, Alan Murchison
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The Canadian Used Fuel Container (UFC) is a mid-size hemispherical headed copper coated steel container measuring 2.5 meters in length and 0.5 meters in diameter containing 48 used fuel bundles. The contained used fuel produces significant gamma radiation requiring automated assembly processes to complete the assembly. The design throughput of 2,500 UFCs per year places constraints on equipment and hot cell design for repeatability, speed of processing, robustness and recovery from upset conditions. After UFC assembly, the UFC is inserted into a Buffer Box (BB). The BB is made from adequately pre-shaped blocks (lower and upper block) and Highly Compacted Bentonite (HCB) material. The blocks are practically ‘sandwiching’ the UFC between them after assembly. This paper identifies one possible approach for the BB automatic assembly cell and processes. Automation of the BB assembly will have a significant positive impact on nuclear safety, quality, productivity, and reliability.Keywords: used fuel packing plant, automatic assembly cell, used fuel container, buffer box, deep geological repository
Procedia PDF Downloads 27523791 Two Dimensional Finite Element Model to Study Calcium Dynamics in Fibroblast Cell with Excess Buffer Approximation Involving ER Flux and SERCA Pump
Authors: Mansha Kotwani
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The specific spatio-temporal calcium concentration patterns are required by the fibroblasts to maintain its structure and functions. Thus, calcium concentration is regulated in cell at different levels in various activities of the cell. The variations in cytosolic calcium concentration largely depend on the buffers present in cytosol and influx of calcium into cytosol from ER through IP3Rs or Raynodine receptors followed by reuptake of calcium into ER through sarcoplasmic/endoplasmic reticulum ATPs (SERCA) pump. In order to understand the mechanisms of wound repair, tissue remodeling and growth performed by fibroblasts, it is of crucial importance to understand the mechanisms of calcium concentration regulation in fibroblasts. In this paper, a model has been developed to study calcium distribution in NRK fibroblast in the presence of buffers and ER flux with SERCA pump. The model has been developed for two dimensional unsteady state case. Appropriate initial and boundary conditions have been framed along with physiology of the cell. Finite element technique has been employed to obtain the solution. The numerical results have been used to study the effect of buffers, ER flux and source amplitude on calcium distribution in fibroblast cell.Keywords: buffers, IP3R, ER flux, SERCA pump, source amplitude
Procedia PDF Downloads 24323790 Preparation of Polymer-Stabilized Magnetic Iron Oxide as Selective Drug Nanocarriers to Human Acute Myeloid Leukemia
Authors: Kheireddine El-Boubbou
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Drug delivery to target human acute myeloid leukemia (AML) using a nanoparticulate chemotherapeutic formulation that can deliver drugs selectively to AML cancer is hugely needed. In this work, we report the development of a nanoformulation made of polymeric-stabilized multifunctional magnetic iron oxide nanoparticles (PMNP) loaded with the anticancer drug Doxorubicin (Dox) as a promising drug carrier to treat AML. Dox@PMNP conjugates simultaneously exhibited high drug content, maximized fluorescence, and excellent release properties. Nanoparticulate uptake and cell death following addition of Dox@PMNPs were then evaluated in different types of human AML target cells, as well as on normal human cells. While the unloaded MNPs were not toxic to any of the cells, Dox@PMNPs were found to be highly toxic to the different AML cell lines, albeit at different inhibitory concentrations (IC50 values), but showed very little toxicity towards the normal cells. In comparison, free Dox showed significant potency concurrently to all the cell lines, suggesting huge potentials for the use of Dox@PMNPs as selective AML anticancer cargos. Live confocal imaging, fluorescence and electron microscopy confirmed that Dox is indeed delivered to the nucleus in relatively short periods of time, causing apoptotic cell death. Importantly, this targeted payload may potentially enhance the effectiveness of the drug in AML patients and may further allow physicians to image leukemic cells exposed to Dox@PMNPs using MRI.Keywords: magnetic nanoparticles, drug delivery, acute myeloid leukemia, iron oxide, cancer nanotherapy
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