Search results for: thin-film solar cell
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4959

Search results for: thin-film solar cell

939 Variable Renewable Energy Droughts in the Power Sector – A Model-based Analysis and Implications in the European Context

Authors: Martin Kittel, Alexander Roth

Abstract:

The continuous integration of variable renewable energy sources (VRE) in the power sector is required for decarbonizing the European economy. Power sectors become increasingly exposed to weather variability, as the availability of VRE, i.e., mainly wind and solar photovoltaic, is not persistent. Extreme events, e.g., long-lasting periods of scarce VRE availability (‘VRE droughts’), challenge the reliability of supply. Properly accounting for the severity of VRE droughts is crucial for designing a resilient renewable European power sector. Energy system modeling is used to identify such a design. Our analysis reveals the sensitivity of the optimal design of the European power sector towards VRE droughts. We analyze how VRE droughts impact optimal power sector investments, especially in generation and flexibility capacity. We draw upon work that systematically identifies VRE drought patterns in Europe in terms of frequency, duration, and seasonality, as well as the cross-regional and cross-technological correlation of most extreme drought periods. Based on their analysis, the authors provide a selection of relevant historical weather years representing different grades of VRE drought severity. These weather years will serve as input for the capacity expansion model for the European power sector used in this analysis (DIETER). We additionally conduct robustness checks varying policy-relevant assumptions on capacity expansion limits, interconnections, and level of sector coupling. Preliminary results illustrate how an imprudent selection of weather years may cause underestimating the severity of VRE droughts, flawing modeling insights concerning the need for flexibility. Sub-optimal European power sector designs vulnerable to extreme weather can result. Using relevant weather years that appropriately represent extreme weather events, our analysis identifies a resilient design of the European power sector. Although the scope of this work is limited to the European power sector, we are confident that our insights apply to other regions of the world with similar weather patterns. Many energy system studies still rely on one or a limited number of sometimes arbitrarily chosen weather years. We argue that the deliberate selection of relevant weather years is imperative for robust modeling results.

Keywords: energy systems, numerical optimization, variable renewable energy sources, energy drought, flexibility

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938 Network Pharmacological Evaluation of Holy Basil Bioactive Phytochemicals for Identifying Novel Potential Inhibitors Against Neurodegenerative Disorder

Authors: Bhuvanesh Baniya

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Alzheimer disease is illnesses that are responsible for neuronal cell death and resulting in lifelong cognitive problems. Due to their unclear mechanism, there are no effective drugs available for the treatment. For a long time, herbal drugs have been used as a role model in the field of the drug discovery process. Holy basil in the Indian medicinal system (Ayurveda) is used for several neuronal disorders like insomnia and memory loss for decades. This study aims to identify active components of holy basil as potential inhibitors for the treatment of Alzheimer disease. To fulfill this objective, the Network pharmacology approach, gene ontology, pharmacokinetics analysis, molecular docking, and molecular dynamics simulation (MDS) studies were performed. A total of 7 active components in holy basil, 12 predicted neurodegenerative targets of holy basil, and 8063 Alzheimer-related targets were identified from different databases. The network analysis showed that the top ten targets APP, EGFR, MAPK1, ESR1, HSPA4, PRKCD, MAPK3, ABL1, JUN, and GSK3B were found as significant target related to Alzheimer disease. On the basis of gene ontology and topology analysis results, APP was found as a significant target related to Alzheimer’s disease pathways. Further, the molecular docking results to found that various compounds showed the best binding affinities. Further, MDS top results suggested could be used as potential inhibitors against APP protein and could be useful for the treatment of Alzheimer’s disease.

Keywords: holy basil, network pharmacology, neurodegeneration, active phytochemicals, molecular docking and simulation

Procedia PDF Downloads 101
937 Na Doped ZnO UV Filters with Reduced Photocatalytic Activity for Sunscreen Application

Authors: Rafid Mueen, Konstantin Konstantinov, Micheal Lerch, Zhenxiang Cheng

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In the past two decades, the concern for skin protection from ultraviolet (UV) radiation has attracted considerable attention due to the increased intensity of UV rays that can reach the Earth’s surface as a result of the breakdown of ozone layer. Recently, UVA has also attracted attention, since, in comparison to UVB, it can penetrate deeply into the skin, which can result in significant health concerns. Sunscreen agents are one of the significant tools to protect the skin from UV irradiation, and it is either organic or in organic. Developing of inorganic UV blockers is essential, which provide efficient UV protection over a wide spectrum rather than organic filters. Furthermore inorganic UV blockers are good comfort, and high safety when applied on human skin. Inorganic materials can absorb, reflect, or scatter the ultraviolet radiation, depending on their particle size, unlike the organic blockers, which absorb the UV irradiation. Nowadays, most inorganic UV-blocking filters are based on (TiO2) and ZnO). ZnO can provide protection in the UVA range. Indeed, ZnO is attractive for in sunscreen formulization, and this relates to many advantages, such as its modest refractive index (2.0), absorption of a small fraction of solar radiation in the UV range which is equal to or less than 385 nm, its high probable recombination of photogenerated carriers (electrons and holes), large direct band gap, high exciton binding energy, non-risky nature, and high tendency towards chemical and physical stability which make it transparent in the visible region with UV protective activity. A significant issue for ZnO use in sunscreens is that it can generate ROS in the presence of UV light because of its photocatalytic activity. Therefore it is essential to make a non-photocatalytic material through modification by other metals. Several efforts have been made to deactivate the photocatalytic activity of ZnO by using inorganic surface modifiers. The doping of ZnO by different metals is another way to modify its photocatalytic activity. Recently, successful doping of ZnO with different metals such as Ce, La, Co, Mn, Al, Li, Na, K, and Cr by various procedures, such as a simple and facile one pot water bath, co-precipitation, hydrothermal, solvothermal, combustion, and sol gel methods has been reported. These materials exhibit greater performance than undoped ZnO towards increasing the photocatalytic activity of ZnO in visible light. Therefore, metal doping can be an effective technique to modify the ZnO photocatalytic activity. However, in the current work, we successfully reduce the photocatalytic activity of ZnO through Na doped ZnO fabricated via sol-gel and hydrothermal methods.

Keywords: photocatalytic, ROS, UVA, ZnO

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936 Food Supplements and Natural Products to Slow Down Biological Aging

Authors: Coppa Federica, Iannello Giulia, Pennisi Stefania, Giuffrida Graziella, Lo Faro Riccardo, Cartelli Simone, Ferruggia Greta, Brundo Maria Violetta

Abstract:

In recent years, a new field of basic research has emerged: the biology and physiology of extracellular vesicles and their application in diagnostics and therapy. In particular, exosomes attract the scientific community as nanovesicles of endosomal origin, which can be secreted by a variety of cells and are found in all biological fluids. Exosomes have recently gained attention also in the cosmetic field: in fact, they are used in creams, serums and masks for topical use, proving to have a series of therapeutic and anti-aging benefits. To date, the oral administration of exosomes is the subject of attention because it represents a non-invasive and efficient method for delivering bioactive molecules into the intestine. We decided to focus our research on the creation of a food supplement that contains various bioactive factors, vitamins, and a new technology called AMPLEX PLUS, containing a mixture of 20 different biologically active factors (GF20) and exosomes isolated and purified from bovine colostrum. We have demonstrated in vitro that this new supplement acts on telomerase, slowing down cell aging. Amplex plus increased the proliferation rate of cells and the addition of it reduced the rate of telomere shortening. Under oxidative stress conditions (H2O2 – induced), the TSR increased; however, treatment with colostrum appeared to attenuate this increase. In particular, after 2 weeks of treatment, AMPLEX plus increased the proliferation rate of cells and exerted a protective effect on telomere length erosion, reducing the rate of its shortening.

Keywords: AMPLEX PLUS, colostrum, exosomes, telomerase

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935 Behavioural Studies on Multidirectional Reinforced 4-D Orthogonal Composites on Various Preform Configurations

Authors: Sriram Venkatesh, V. Murali Mohan, T. V. Karthikeyan

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The main advantage of multi-directionally reinforced composites is the freedom to orient selected fibre types and hence derives the benefits of varying fibre volume fractions and there by accommodate the design loads of the final structure of composites. This technology provides the means to produce tailored composites with desired properties. Due to the high level of fibre integrity with through thickness reinforcement those composites are expected to exhibit superior load bearing characteristics with capability to carry load even after noticeable and apparent fracture. However a survey of published literature indicates inadequacy in the design and test data base for the complete characterization of the multidirectional composites. In this paper the research objective is focused on the development and testing of 4-D orthogonal composites with different preform configurations and resin systems. A preform is the skeleton 4D reinforced composite other than the matrix. In 4-D preforms fibre bundles are oriented in three directions at 1200 with respect to each other and they are on orthogonal plane with the fibre in 4th direction. This paper addresses the various types of 4-D composite manufacturing processes and the mechanical test methods followed for the material characterization. A composite analysis is also made, experiments on course and fine woven preforms are conducted and the findings of test results are discussed in this paper. The interpretations of the test results reveal several useful and interesting features. This should pave the way for more widespread use of the perform configurations for allied applications.

Keywords: multi-directionally reinforced composites, 4-D orthogonal preform, course weave, fine weave, fibre bundle spools, unit cell, fibre architecture, fibre volume fraction, fibre distribution

Procedia PDF Downloads 233
934 Electrochemical Recovery of Lithium from Geothermal Brines

Authors: Sanaz Mosadeghsedghi, Mathew Hudder, Mohammad Ali Baghbanzadeh, Charbel Atallah, Seyedeh Laleh Dashtban Kenari, Konstantin Volchek

Abstract:

Lithium has recently been extensively used in lithium-ion batteries (LIBs) for electric vehicles and portable electronic devices. The conventional evaporative approach to recover and concentrate lithium is extremely slow and may take 10-24 months to concentrate lithium from dilute sources, such as geothermal brines. To response to the increasing industrial lithium demand, alternative extraction and concentration technologies should be developed to recover lithium from brines with low concentrations. In this study, a combination of electrocoagulation (EC) and electrodialysis (ED) was evaluated for the recovery of lithium from geothermal brines. The brine samples in this study, collected in Western Canada, had lithium concentrations of 50-75 mg/L on a background of much higher (over 10,000 times) concentrations of sodium. This very high sodium-to-lithium ratio poses challenges to the conventional direct-lithium extraction processes which employ lithium-selective adsorbents. EC was used to co-precipitate lithium using a sacrificial aluminium electrode. The precipitate was then dissolved, and the leachate was treated using ED to separate and concentrate lithium from other ions. The focus of this paper is on the study of ED, including a two-step ED process that included a mono-valent selective stage to separate lithium from multi-valent cations followed by a bipolar ED stage to convert lithium chloride (LiCl) to LiOH product. Eventually, the ED cell was reconfigured using mono-valent cation exchange with the bipolar membranes to combine the two ED steps in one. Using this process at optimum conditions, over 95% of the co-existing cations were removed and the purity of lithium increased to over 90% in the final product.

Keywords: electrochemical separation, electrocoagulation, electrodialysis, lithium extraction

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933 Design of Bacterial Pathogens Identification System Based on Scattering of Laser Beam Light and Classification of Binned Plots

Authors: Mubashir Hussain, Mu Lv, Xiaohan Dong, Zhiyang Li, Bin Liu, Nongyue He

Abstract:

Detection and classification of microbes have a vast range of applications in biomedical engineering especially in detection, characterization, and quantification of bacterial contaminants. For identification of pathogens, different techniques are emerging in the field of biomedical engineering. Latest technology uses light scattering, capable of identifying different pathogens without any need for biochemical processing. Bacterial Pathogens Identification System (BPIS) which uses a laser beam, passes through the sample and light scatters off. An assembly of photodetectors surrounded by the sample at different angles to detect the scattering of light. The algorithm of the system consists of two parts: (a) Library files, and (b) Comparator. Library files contain data of known species of bacterial microbes in the form of binned plots, while comparator compares data of unknown sample with library files. Using collected data of unknown bacterial species, highest voltage values stored in the form of peaks and arranged in 3D histograms to find the frequency of occurrence. Resulting data compared with library files of known bacterial species. If sample data matching with any library file of known bacterial species, sample identified as a matched microbe. An experiment performed to identify three different bacteria particles: Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia coli. By applying algorithm using library files of given samples, results were compromising. This system is potentially applicable to several biomedical areas, especially those related to cell morphology.

Keywords: microbial identification, laser scattering, peak identification, binned plots classification

Procedia PDF Downloads 150
932 Synthesis, Characterization and Catecholase Study of Novel Bidentate Schiff Base Derived from Dehydroacetic Acid

Authors: Salima Tabti, Chaima Maouche, Tinhinene Louaileche, Amel Djedouani, Ismail Warad

Abstract:

Novel Schiff base ligand HL has been synthesized by condensation of aromatic amine and DHA. It was characterized by UV-Vis, FT-IR, SM, NMR (1H, 13C) and also by single-crystal X-ray diffraction. The crystal structure shows that compound crystallized in a triclinic system in P-1 space group and with a two unit per cell (Z = 2).The asymmetric unit, contains one independent molecules, the conformation is determined by an intermolecular N-H…O hydrogen bond with an S(6) ring motif. The molecule have an (E) conformation about the C=N bond. The dihedral angles between the phenyl and pyran ring planes is 89.37 (1), the two plans are approximately perpendicular. The catecholase activity of is situ copper complexes of this ligand has been investigated against catechol. The progress of the oxidation reactions was closely monitored over time following the strong peak of catechol using UV-Vis. Oxidation rates were determined from the initial slope of absorbance. time plots, then analyzed by Michaelis-Menten equations. Catechol oxidation reactions were realized using different concentrations of copper acetate and ligand (L/Cu: 1/1, 1/2, 2/1). The results show that all complexes were able to catalyze the oxidation of catechol. Acetate complexes have the highest activity. Catalysis is a branch of chemical kinetics that, more generally, studies the influence of all physical or chemical factors determining reaction rates. It solves a lot of problems in the chemistry reaction process, especially for a green, economic and less polluting chemistry. For this reason, the search for new catalysts for known organic reactions, occupies a very advanced place in the themes proposed by the chemists.

Keywords: dehydroacetic acid, catechol, copper, catecholase activity, x-ray

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931 Quantitative Proteome Analysis and Bioactivity Testing of New Zealand Honeybee Venom

Authors: Maryam Ghamsari, Mitchell Nye-Wood, Kelvin Wang, Angela Juhasz, Michelle Colgrave, Don Otter, Jun Lu, Nazimah Hamid, Thao T. Le

Abstract:

Bee venom, a complex mixture of peptides, proteins, enzymes, and other bioactive compounds, has been widely studied for its therapeutic application. This study investigated the proteins present in New Zealand (NZ) honeybee venom (BV) using bottom-up proteomics. Two sample digestion techniques, in-solution digestion and filter-aided sample preparation (FASP), were employed to obtain the optimal method for protein digestion. Sequential Window Acquisition of All Theoretical Mass Spectra (SWATH–MS) analysis was conducted to quantify the protein compositions of NZ BV and investigate variations in collection years. Our results revealed high protein content (158.12 µg/mL), with the FASP method yielding a larger number of identified proteins (125) than in-solution digestion (95). SWATH–MS indicated melittin and phospholipase A2 as the most abundant proteins. Significant variations in protein compositions across samples from different years (2018, 2019, 2021) were observed, with implications for venom's bioactivity. In vitro testing demonstrated immunomodulatory and antioxidant activities, with a viable range for cell growth established at 1.5-5 µg/mL. The study underscores the value of proteomic tools in characterizing bioactive compounds in bee venom, paving the way for deeper exploration into their therapeutic potentials. Further research is needed to fractionate the venom and elucidate the mechanisms of action for the identified bioactive components.

Keywords: honeybee venom, proteomics, bioactivity, fractionation, swath-ms, melittin, phospholipase a2, new zealand, immunomodulatory, antioxidant

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930 Complex Management of Arrhythmogenic Right Ventricular Dysplasia/Cardiomyopathy

Authors: Fahad Almehmadi, Abdullah Alrajhi, Bader K. Alaslab, Abdullah A. Al Qurashi, Hattan A. Hassani

Abstract:

Arrhythmogenic Right Ventricular Dysplasia/Cardiomyopathy (ARVD/C) is an uncommon, inheritable cardiac disorder characterized by the progressive substitution of cardiac myocytes by fibro-fatty tissues. This pathologic substitution predisposes patients to ventricular arrhythmias and right ventricular failure. The underlying genetic defect predominantly involves genes encoding for desmosome proteins, particularly plakophilin-2 (PKP2). These aberrations lead to impaired cell adhesion, heightening the susceptibility to fibrofatty scarring under conditions of mechanical stress. Primarily, ARVD/C affects the right ventricle, but it can also compromise the left ventricle, potentially leading to biventricular heart failure. Clinical presentations can vary, spanning from asymptomatic individuals to those experiencing palpitations, syncopal episodes, and, in severe instances, sudden cardiac death. The establishment of a diagnostic criterion specifically tailored for ARVD/C significantly aids in its accurate diagnosis. Nevertheless, the task of early diagnosis is complicated by the disease's frequently asymptomatic initial stages, and the overall rarity of ARVD/C cases reported globally. In some cases, as exemplified by the adult female patient in this report, the disease may advance to terminal stages, rendering therapies like Ventricular Tachycardia (VT) ablation ineffective. This case underlines the necessity for increased awareness and understanding of ARVD/C to aid in its early detection and management. Through such efforts, we aim to decrease morbidity and mortality associated with this challenging cardiac disorder.

Keywords: ARVD/C, cardiology, interventional cardiology, cardiac electrophysiology

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929 Evaluation of Electro-Flocculation for Biomass Production of Marine Microalgae Phaodactylum tricornutum

Authors: Luciana C. Ramos, Leandro J. Sousa, Antônio Ferreira da Silva, Valéria Gomes Oliveira Falcão, Suzana T. Cunha Lima

Abstract:

The commercial production of biodiesel using microalgae demands a high-energy input for harvesting biomass, making production economically unfeasible. Methods currently used involve mechanical, chemical, and biological procedures. In this work, a flocculation system is presented as a cost and energy effective process to increase biomass production of Phaeodactylum tricornutum. This diatom is the only species of the genus that present fast growth and lipid accumulation ability that are of great interest for biofuel production. The algae, selected from the Bank of Microalgae, Institute of Biology, Federal University of Bahia (Brazil), have been bred in tubular reactor with photoperiod of 12 h (clear/dark), providing luminance of about 35 μmol photons m-2s-1, and temperature of 22 °C. The medium used for growing cells was the Conway medium, with addition of silica. The seaweed growth curve was accompanied by cell count in Neubauer camera and by optical density in spectrophotometer, at 680 nm. The precipitation occurred at the end of the stationary phase of growth, 21 days after inoculation, using two methods: centrifugation at 5000 rpm for 5 min, and electro-flocculation at 19 EPD and 95 W. After precipitation, cells were frozen at -20 °C and, subsequently, lyophilized. Biomass obtained by electro-flocculation was approximately four times greater than the one achieved by centrifugation. The benefits of this method are that no addition of chemical flocculants is necessary and similar cultivation conditions can be used for the biodiesel production and pharmacological purposes. The results may contribute to improve biodiesel production costs using marine microalgae.

Keywords: biomass, diatom, flocculation, microalgae

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928 In-Vitro Dextran Synthesis and Characterization of an Intracellular Glucosyltransferase from Leuconostoc Mesenteroides AA1

Authors: Afsheen Aman, Shah Ali Ul Qader

Abstract:

Dextransucrase [EC 2.4.1.5] is a glucosyltransferase that catalysis the biosynthesis of a natural biopolymer called dextran. It can catalyze the transfer of D-glucopyranosyl residues from sucrose to the main chain of dextran. This unique biopolymer has multiple applications in several industries and the key utilization of dextran lies on its molecular weight and the type of branching. Extracellular dextransucrase from Leuconostoc mesenteroides is most extensively studied and characterized. Limited data is available regarding cell-bound or intracellular dextransucrase and on the characterization of dextran produced by in-vitro reaction of intracellular dextransucrase. L. mesenteroides AA1 is reported to produce extracellular dextransucrase that catalyzes biosynthesis of a high molecular weight dextran with only α-(1→6) linkage. Current study deals with the characterization of an intracellular dextransucrase and in vitro biosynthesis of low molecular weight dextran from L. mesenteroides AA1. Intracellular dextransucrase was extracted from cytoplasm and purified to homogeneity for characterization. Kinetic constants, molecular weight and N-terminal sequence analysis of intracellular dextransucrase reveal unique variation with previously reported extracellular dextransucrase from the same strain. In vitro synthesized biopolymer was characterized using NMR spectroscopic techniques. Intracellular dextransucrase exhibited Vmax and Km values of 130.8 DSU ml-1 hr-1 and 221.3 mM, respectively. Optimum catalytic activity was detected at 35°C in 0.15 M citrate phosphate buffer (pH-5.5) in 05 minutes. Molecular mass of purified intracellular dextransucrase is approximately 220.0 kDa on SDS-PAGE. N-terminal sequence of the intracellular enzyme is: GLPGYFGVN that showed no homology with previously reported sequence for the extracellular dextransucrase. This intracellular dextransucrase is capable of in vitro synthesis of dextran under specific conditions. This intracellular dextransucrase is capable of in vitro synthesis of dextran under specific conditions and this biopolymer can be hydrolyzed into different molecular weight fractions for various applications.

Keywords: characterization, dextran, dextransucrase, leuconostoc mesenteroides

Procedia PDF Downloads 396
927 Synergistic Effect of Doxorubicin-Loaded Silver Nanoparticles – Polymeric Conjugates on Breast Cancer Cells

Authors: Nancy M. El-Baz, Laila Ziko, Rania Siam, Wael Mamdouh

Abstract:

Cancer is one of the most devastating diseases, and has over than 10 million new cases annually worldwide. Despite the effectiveness of chemotherapeutic agents, their systemic toxicity and non-selective anticancer actions represent the main obstacles facing cancer curability. Due to the effective enhanced permeability and retention (EPR) effect of nanomaterials, nanoparticles (NPs) have been used as drug nanocarriers providing targeted cancer drug delivery systems. In addition, several inorganic nanoparticles such as silver (AgNPs) nanoparticles demonstrated a potent anticancer activity against different cancers. The present study aimed at formulating core-shell inorganic NPs-based combinatorial therapy based on combining the anticancer activity of AgNPs along with doxorubicin (DOX) and evaluating their cytotoxicity on MCF-7 breast cancer cells. These inorganic NPs-based combinatorial therapies were designed to (i) Target and kill cancer cells with high selectivity, (ii) Have an improved efficacy/toxicity balance, and (iii) Have an enhanced therapeutic index when compared to the original non-modified DOX with much lower dosage The in-vitro cytotoxicity studies demonstrated that the NPs-based combinatorial therapy achieved the same efficacy of non-modified DOX on breast cancer cell line, but with 96% reduced dose. Such reduction in DOX dose revealed that the combination between DOX and NPs possess a synergic anticancer activity against breast cancer. We believe that this is the first report on a synergic anticancer effect at very low dose of DOX against MCF-7 cells. Future studies on NPs-based combinatorial therapy may aid in formulating novel and significantly more effective cancer therapeutics.

Keywords: nanoparticles-based combinatorial therapy, silver nanoparticles, doxorubicin, breast cancer

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926 The Localization and Function of p38α Mitogen-Activated Protein Kinase (MAPK) in Rat Oocytes

Authors: Shifu Hu, Qiong Yu, Wei Xia, Changhong Zhu

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Background: P38α MAPK, which is a member of the canonical MAPK family, is activated in response to various extracellular stresses and plays a role in multiple cellular processes. It is well known that p38α MAPK play vital roles in oocyte maturation, but the localization and functional roles of p38α MAPK during the meiotic maturation of rat oocytes remain unknown. Study Design: In this study, western-blot and immunofluorescent staining were used to investigate the expression and subcellular localization of p38α MAPK during the meiotic maturation of rat oocytes. SB203580, a specific inhibitor of p38α MAPK, was used to study the roles of p38α MAPK in the meiotic cell cycle of rat oocytes. Results: The results found that p38α MAPK phosphorylation (p-p38α MAPK, indicative of p38α MAPK activation) was low at the germinal vesicle (GV) stage, increased 3 h after germinal vesicle breakdown (GVBD), and maintained its maximum at MI (metaphase I) or M II (metaphase II). The p-p38α MAPK mainly accumulated in the germinal vesicle and had no obvious expression in the nucleus. From GVBD to M II, p-p38α MAPK was distributed in the cytoplasm around either the chromosomes or the spindle. We used SB203580, an inhibitor of p38α MAPK, to investigate the possible functional role of p38α MAPK during rat oocyte meiotic maturation. Treatment of GV stage oocytes with 20 μM SB203580 blocked p-p38α MAPK activity, and the spindles appeared abnormal. Additionally, the rate of GVBD after 3h of culture with 20 μM SB203580 (58.8%) was significantly inhibited compared with the control (82.5%, p < 0.05), and the polar body extrusion rate after 12 h of culture with SB203580 was also significantly decreased compared with the control (40.1 vs. 73.3%, p < 0.05). Conclusions: These data indicate that p38α MAPK may play a vital role in rat oocyte meiotic maturation.

Keywords: meiotic maturation, oocyte, p38α MAPK, spindle

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925 A Sub-Conjunctiva Injection of Rosiglitazone for Anti-Fibrosis Treatment after Glaucoma Filtration Surgery

Authors: Yang Zhao, Feng Zhang, Xuanchu Duan

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Trans-differentiation of human Tenon fibroblasts (HTFs) to myo-fibroblasts and fibrosis of episcleral tissue are the most common reasons for the failure of glaucoma filtration surgery, with limited treatment options like antimetabolites which always have side-effects such as leakage of filter bulb, infection, hypotony, and endophthalmitis. Rosiglitazone, a specific thiazolidinedione is a synthetic high-affinity ligand for PPAR-r, which has been used in the treatment of type2 diabetes, and found to have pleiotropic functions against inflammatory response, cell proliferation and tissue fibrosis and to benefit to a variety of diseases in animal myocardium models, steatohepatitis models, etc. Here, in vitro we cultured primary HTFs and stimulated with TGF- β to induced myofibrogenic, then treated cells with Rosiglitazone to assess for fibrogenic response. In vivo, we used rabbit glaucoma model to establish the formation of post- trabeculectomy scarring. Then we administered subconjunctival injection with Rosiglitazone beside the filtering bleb, later protein, mRNA and immunofluorescence of fibrogenic markers are checked, and filtering bleb condition was measured. In vitro, we found Rosiglitazone could suppressed proliferation and migration of fibroblasts through macroautophagy via TGF- β /Smad signaling pathway. In vivo, on postoperative day 28, the mean number of fibroblasts in Rosiglitazone injection group was significantly the lowest and had the least collagen content and connective tissue growth factor. Rosiglitazone effectively controlled human and rabbit fibroblasts in vivo and in vitro. Its subconjunctiiva application may represent an effective, new avenue for the prevention of scarring after glaucoma surgery.

Keywords: fibrosis, glaucoma, macroautophagy, rosiglitazone

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924 [Keynote Talk]: Mathematical and Numerical Modelling of the Cardiovascular System: Macroscale, Mesoscale and Microscale Applications

Authors: Aymen Laadhari

Abstract:

The cardiovascular system is centered on the heart and is characterized by a very complex structure with different physical scales in space (e.g. micrometers for erythrocytes and centimeters for organs) and time (e.g. milliseconds for human brain activity and several years for development of some pathologies). The development and numerical implementation of mathematical models of the cardiovascular system is a tremendously challenging topic at the theoretical and computational levels, inducing consequently a growing interest over the past decade. The accurate computational investigations in both healthy and pathological cases of processes related to the functioning of the human cardiovascular system can be of great potential in tackling several problems of clinical relevance and in improving the diagnosis of specific diseases. In this talk, we focus on the specific task of simulating three particular phenomena related to the cardiovascular system on the macroscopic, mesoscopic and microscopic scales, respectively. Namely, we develop numerical methodologies tailored for the simulation of (i) the haemodynamics (i.e., fluid mechanics of blood) in the aorta and sinus of Valsalva interacting with highly deformable thin leaflets, (ii) the hyperelastic anisotropic behaviour of cardiomyocytes and the influence of calcium concentrations on the contraction of single cells, and (iii) the dynamics of red blood cells in microvasculature. For each problem, we present an appropriate fully Eulerian finite element methodology. We report several numerical examples to address in detail the relevance of the mathematical models in terms of physiological meaning and to illustrate the accuracy and efficiency of the numerical methods.

Keywords: finite element method, cardiovascular system, Eulerian framework, haemodynamics, heart valve, cardiomyocyte, red blood cell

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923 Prediction of Antibacterial Peptides against Propionibacterium acnes from the Peptidomes of Achatina fulica Mucus Fractions

Authors: Suwapitch Chalongkulasak, Teerasak E-Kobon, Pramote Chumnanpuen

Abstract:

Acne vulgaris is a common skin disease mainly caused by the Gram–positive pathogenic bacterium, Propionibacterium acnes. This bacterium stimulates inflammation process in human sebaceous glands. Giant African snail (Achatina fulica) is alien species that rapidly reproduces and seriously damages agricultural products in Thailand. There were several research reports on the medical and pharmaceutical benefits of this snail mucus peptides and proteins. This study aimed to in silico predict multifunctional bioactive peptides from A. fulica mucus peptidome using several bioinformatic tools for determination of antimicrobial (iAMPpred), anti–biofilm (dPABBs), cytotoxic (Toxinpred), cell membrane penetrating (CPPpred) and anti–quorum sensing (QSPpred) peptides. Three candidate peptides with the highest predictive score were selected and re-designed/modified to improve the required activities. Structural and physicochemical properties of six anti–P. acnes (APA) peptide candidates were performed by PEP–FOLD3 program and the five aforementioned tools. All candidates had random coiled structure and were named as APA1–ori, APA2–ori, APA3–ori, APA1–mod, APA2–mod and APA3–mod. To validate the APA activity, these peptide candidates were synthesized and tested against six isolates of P. acnes. The modified APA peptides showed high APA activity on some isolates. Therefore, our biomimetic mucus peptides could be useful for preventing acne vulgaris and further examined on other activities important to medical and pharmaceutical applications.

Keywords: Propionibacterium acnes, Achatina fulica, peptidomes, antibacterial peptides, snail mucus

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922 Anti-Prostate Cancer Effect of GV-1001, a Novel Gonadotropin-Releasing Hormone Receptor Ligand

Authors: Ji Won Kim, Moo Yeol Lee, Keon Wook Kang

Abstract:

GV-1001, 16 amino acid fragment of human telomerase reverse transcriptase catalytic subunit (hTERT), has been developed as an injectable cancer vaccine for many types of solid tumors showing high-level of telomerase activity. In the present study, we evaluated the anti-cancer effect of GV-1001 on androgen-receptor-positive prostate cancer. Two signaling pathways, Gs-adenylate cyclase-cAMP and Gq-IP3-Ca2+ pathways play a central role in GnRH receptor (GnRHR)-mediated activities. We found that leuprolide acetate (LA) mainly acted on Gq-mediated Ca2+ signaling, while GV-1001 preferentially acted on cAMP signaling; and both the effects were counteracted by cetrorelix, a GnRHR antagonist. We further tested whether GV-1001 affects tumor growth of human prostate cancer cells in vivo. Prostate tumor xenografts were established using LNCap, androgen receptor-positive prostate cancer cells, and the nude mice bearing tumors were subcutaneously injected with GV-1001 (0.01, 0.1, 1, 10 microg/kg/day) and LA (0.01 microg/kg/day) for 2 weeks. GV-1001 (1 and 10 microg/kg/day) significantly inhibited tumor growth of LNCap xenografts. Interestingly, mRNA expression of MMP2 and MMP9 was significantly suppressed by GV-1001 injection, but not by LA administration. Boyden chamber assay revealed that GV-1001 potently inhibited cell migration of LNCap. Our finding suggests that GV-1001 as a novel GnRHR ligand, has anti-proliferative and anti-migratory effects on androgen receptor-positive prostate cancer cells.

Keywords: GV-1001, GnRH, hTERT, prostate cancer

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921 Nanoparticles Modification by Grafting Strategies for the Development of Hybrid Nanocomposites

Authors: Irati Barandiaran, Xabier Velasco-Iza, Galder Kortaberria

Abstract:

Hybrid inorganic/organic nanostructured materials based on block copolymers are of considerable interest in the field of Nanotechnology, taking into account that these nanocomposites combine the properties of polymer matrix and the unique properties of the added nanoparticles. The use of block copolymers as templates offers the opportunity to control the size and the distribution of inorganic nanoparticles. This research is focused on the surface modification of inorganic nanoparticles to reach a good interface between nanoparticles and polymer matrices which hinders the nanoparticle aggregation. The aim of this work is to obtain a good and selective dispersion of Fe3O4 magnetic nanoparticles into different types of block copolymers such us, poly(styrene-b-methyl methacrylate) (PS-b-PMMA), poly(styrene-b-ε-caprolactone) (PS-b-PCL) poly(isoprene-b-methyl methacrylate) (PI-b-PMMA) or poly(styrene-b-butadiene-b-methyl methacrylate) (SBM) by using different grafting strategies. Fe3O4 magnetic nanoparticles have been surface-modified with polymer or block copolymer brushes following different grafting methods (grafting to, grafting from and grafting through) to achieve a selective location of nanoparticles into desired domains of the block copolymers. Morphology of fabricated hybrid nanocomposites was studied by means of atomic force microscopy (AFM) and with the aim to reach well-ordered nanostructured composites different annealing methods were used. Additionally, nanoparticle amount has been also varied in order to investigate the effect of the nanoparticle content in the morphology of the block copolymer. Nowadays different characterization methods were using in order to investigate magnetic properties of nanometer-scale electronic devices. Particularly, two different techniques have been used with the aim of characterizing synthesized nanocomposites. First, magnetic force microscopy (MFM) was used to investigate qualitatively the magnetic properties taking into account that this technique allows distinguishing magnetic domains on the sample surface. On the other hand, magnetic characterization by vibrating sample magnetometer and superconducting quantum interference device. This technique demonstrated that magnetic properties of nanoparticles have been transferred to the nanocomposites, exhibiting superparamagnetic behavior similar to that of the maghemite nanoparticles at room temperature. Obtained advanced nanostructured materials could found possible applications in the field of dye-sensitized solar cells and electronic nanodevices.

Keywords: atomic force microscopy, block copolymers, grafting techniques, iron oxide nanoparticles

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920 Multi-Modality Imaging of Aggressive Hoof Wall Neoplasia in Two Horses

Authors: Hannah Nagel, Hayley Lang, Albert Sole Guitart, Natasha Lean, Rachel Allavena, Cleide Sprohnie-Barrera, Alex Young

Abstract:

Aggressive neoplasia of the hoof is a rare occurrence in horses and has been only sporadically described in the literature. In the few cases reported intra-hoof wall, aggressive neoplasia has been documented radiographically and has been described with variable imaging characteristics. These include a well-defined osteolytic area, a smoothly outlined semi-circular defect, an extensive draining tract beneath the hoof wall, as well as an additional large area of osteolysis or an extensive central lytic region. A 20-year-old Quarterhorse gelding and a 10-year-old Thoroughbred gelding were both presented for chronic reoccurring lameness in the left forelimb and left hindlimb, respectively. Both of the cases displayed radiographic lesions that have been previously described but also displayed osteoproliferative expansile regions of additional bone formation. Changes associated with hoof neoplasia are often non-specific due to the nature and capacity of bone to react to pathological insult, which is either to proliferate or be absorbed. Both cases depict and describe imaging findings seen on radiography, contrast radiography, computed tomography, and magnetic resonance imaging before reaching a histological diagnosis of malignant melanoma and squamous cell carcinoma. Although aggressive hoof wall neoplasia is rare, there are some imaging features which may raise our index of suspicion for an aggressive hoof wall lesion. This case report documents two horses with similar imaging findings who underwent multiple assessments, surgical interventions, and imaging modalities with a final diagnosis of malignant neoplasia.

Keywords: horse, hoof, imaging, radiography, neoplasia

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919 Psychological Nano-Therapy: A New Method in Family Therapy

Authors: Siamak Samani, Nadereh Sohrabi

Abstract:

Psychological nano-therapy is a new method based on systems theory. According to the theory, systems with severe dysfunctions are resistant to changes. Psychological nano-therapy helps the therapists to break this ice. Two key concepts in psychological nano-therapy are nano-functions and nano-behaviors. The most important step in psychological nano-therapy in family therapy is selecting the most effective nano-function and nano-behavior. The aim of this study was to check the effectiveness of psychological nano-therapy for family therapy. One group pre-test-post-test design (quasi-experimental Design) was applied for research. The sample consisted of ten families with severe marital conflict. The important character of these families was resistance for participating in family therapy. In this study, sending respectful (nano-function) text massages (nano-behavior) with cell phone were applied as a treatment. Cohesion/respect sub scale from self-report family processes scale and family readiness for therapy scale were used to assess all family members in pre-test and post-test. In this study, one of family members was asked to send a respectful text massage to other family members every day for a week. The content of the text massages were selected and checked by therapist. To compare the scores of families in pre-test and post-test paired sample t-test was used. The results of the test showed significant differences in both cohesion/respect score and family readiness for therapy between per-test and post-test. The results revealed that these families have found a better atmosphere for participation in a complete family therapy program. Indeed, this study showed that psychological nano-therapy is an effective method to make family readiness for therapy.

Keywords: family therapy, family conflicts, nano-therapy, family readiness

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918 Sensitivity of Acanthamoeba castellanii-Grown Francisella to Three Different Disinfectants

Authors: M. Knezevic, V. Marecic, M. Ozanic, I. Kelava, M. Mihelcic, M. Santic

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Francisella tularensis is a highly infectious, gram-negative intracellular bacterium and the causative agent of tularemia. The bacterium has been isolated from more than 250 wild species, including protozoa cells. Since Francisella is very virulent and persists in the environment for years, the aim of this study was to investigate whether Acanthamoeba castellanii-grown F. novicida exhibits an alteration in the resistance to disinfectants. It has been shown by other intracellular pathogens, including Legionella pneumophila that bacteria grown in amoeba exhibit more resistance to disinfectants. However, there is no data showing Francisella viability behaviour after intracellular life cycle in A. castellani. In this study, the bacterial suspensions of A. castellanii-grown or in vitro-grown Francisella were treated with three different disinfectants, and the bacterial viability after disinfection treatment was determined by a colony-forming unit (CFU) counting method, transmission electron microscopy (TEM), fluorescence microscopy as well as the leakage of intracellular fluid. Our results have shown that didecyldimethylammonium chloride (DDAC) combined with isopropyl alcohol was the most effective in bacterial killing; all in vitro-grown and A. castellanii-grown F. novicida were killed after only 10s. Surprisingly, in comparison to in vitro-grown bacteria, A. castellanii-grown F. novicida was more sensitive to decontamination by the benzalkonium chloride combined with DDAC and formic acid and the polyhexamethylene biguanide (PHMB). We can conclude that the tested disinfectants exhibit antimicrobial activity by causing a loss of structural organization and integrity of the Francisella cell wall and membrane and the subsequent leakage of the intracellular contents. Finally, the results of this study clearly demonstrate that Francisella grown in A. castellanii had become more susceptible to many disinfectants.

Keywords: Acanthamoeba, disinfectant, Francisella, sensitivity

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917 Analysis of a Faience Enema Found in the Assasif Tomb No. -28- of the Vizier Amenhotep Huy: Contributions to the Study of the Mummification Ritual Practiced in the Theban Necropolis

Authors: Alberto Abello Moreno-Cid

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Mummification was the process through which immortality was granted to the deceased, so it was of extreme importance to the Egyptians. The techniques of embalming had evolved over the centuries, and specialists created increasingly sophisticated tools. However, due to its eminently religious nature, knowledge about everything related to this practice was jealously preserved, and the testimonies that have survived to our time are scarce. For this reason, embalming instruments found in archaeological excavations are uncommon. The tomb of the Vizier Amenhotep Huy (AT No. -28-), located in the el-Assasif necropolis that is being excavated since 2009 by the team of the Institute of Ancient Egyptian Studies, has been the scene of some discoveries of this type that evidences the existence of mummification practices in this place after the New Kingdom. The clysters or enemas are the fundamental tools in the second type of mummification described by the historian Herodotus to introduce caustic solutions inside the body of the deceased. Nevertheless, such objects only have been found in three locations: the tomb of Ankh-Hor in Luxor, where a copper enema belonged to the prophet of Ammon Uah-ib-Ra came to light; the excavation of the tomb of Menekh-ib-Nekau in Abusir, where was also found one made of copper; and the excavations in the Bucheum, where two more artifacts were discovered, also made of copper but in different shapes and sizes. Both of them were used for the mummification of sacred animals and this is the reason they vary significantly. Therefore, the object found in the tomb No. -28-, is the first known made of faience of all these peculiar tools and the oldest known until now, dated in the Third Intermediate Period (circa 1070-650 B.C.). This paper bases its investigation on the study of those parallelisms, the material, the current archaeological context and the full analysis and reconstruction of the object in question. The key point is the use of faience in the production of this item: creating a device intended to be in constant use seems to be a first illogical compared to other samples made of copper. Faience around the area of Deir el-Bahari had a strong religious component, associated with solar myths and principles of the resurrection, connected to the Osirian that characterises the mummification procedure. The study allows to refute some of the premises which are held unalterable in Egyptology, verifying the utilization of these sort of pieces, understanding its way of use and showing that this type of mummification was also applied to the highest social stratum, in which case the tools were thought out of an exceptional quality and religious symbolism.

Keywords: clyster, el-Assasif, embalming, faience enema mummification, Theban necropolis

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916 Evaluation of Cytotoxic Effect of Mitoxantrone Conjugated Magnetite Nanoparticles and Graphene Oxide-Magnetite Nanocomposites on Mesenchymal Stem Cells

Authors: Abbas Jafarizad, Duygu Ekinci

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In this work targeted drug delivery is proposed to decrease adverse effect of drugs with concomitant reduces in consumption and treatment outgoings. Nanoparticles (NPs) can be prepared from a variety of materials such as lipid, biodegradable polymer that prevent the drugs cytotoxicity in healthy cells, etc. One of the most important drugs used in chemotherapy is mitoxantrone (MTX) which prevents cell proliferation by inhibition of topoisomerase II and DNA repair; however, it is not selective and has some serious side effects. In this study, mentioned aim is achieved by using several nanocarriers like magnetite (Fe3O4) and their composites with magnetic graphene oxide (Fe3O4@GO). Also, cytotoxic potential of Fe3O4, Fe3O4-MTX, and Fe3O4@GO-MTX nanocomposite were evaluated on mesenchymal stem cells (MSCs). In this study, we reported the synthesis of monodisperse Fe3O4 NPs and Fe3O4@GO nanocomposite and their structures were investigated by using field emission scanning electron microscope (FESEM), Fourier transform infrared (FTIR) spectra, atomic force microscopy (AFM), Brauneur Emmet Teller (BET) isotherm and contact angle studies. Moreover, we used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to evaluate cytotoxic effects of MTX, Fe3O4 NPs, Fe3O4-MTX and Fe3O4@GO-MTX nanocomposite on MSCs. The in-vitro MTT results indicated that all concentrations of MTX and Fe3O4@GO-MTX nanocomposites showed cytotoxic effects while all concentrations of Fe3O4 NPs and Fe3O4-MTX NPs did not show any cytotoxic effect on stem cells. The results from this study indicated that using Fe3O4 NPs as anticancer drug delivery systems could be a trustworthy method for cancer treatment. But for reaching excellent and accurate results, further investigation is necessary.

Keywords: mitoxantrone, magnetite, magnetic graphene oxide, MTT assay, mesenchymal stem cells

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915 DNA Prime/MVTT Boost Enhances Broadly Protective Immune Response against Mosaic HIV-1 Gag

Authors: Wan Liu, Haibo Wang, Cathy Huang, Zhiwu Tan, Zhiwei Chen

Abstract:

The tremendous diversity of HIV-1 has been a major challenge for an effective AIDS vaccine development. Mosaic approach presents the potential for vaccine design aiming for global protection. The mosaic antigen of HIV-1 Gag allows antigenic breadth for vaccine-elicited immune response against a wider spectrum of viral strains. However, the enhancement of immune response using vaccines is dependent on the strategy used. Heterologous prime/boost regimen has been shown to elicit high levels of immune responses. Here, we investigated whether priming using plasmid DNA with electroporation followed by boosting with the live replication-competent modified vaccinia virus vector TianTan (MVTT) combined with the mosaic antigenic sequence could elicit a greater and broader antigen-specific response against HIV-1 Gag in mice. When compared to DNA or MVTT alone, or MVTT/MVTT group, DNA/MVTT group resulted in coincidentally high frequencies of broadly reactive, Gag-specific, polyfunctional, long-lived, and cytotoxic CD8+ T cells and increased anti-Gag antibody titer. Meanwhile, the vaccination could upregulate PD-1+, and Tim-3+ CD8+ T cell, myeloid-derived suppressive cells and Treg cells to balance the stronger immune response induced. Importantly, the prime/boost vaccination could help control the EcoHIV and mesothelioma AB1-gag challenge. The stronger protective Gag-specific immunity induced by a Mosaic DNA/MVTT vaccine corroborate the promise of the mosaic approach, and the potential of two acceptably safe vectors to enhance anti-HIV immunity and cancer prevention.

Keywords: DNA/MVTT vaccine, EcoHIV, mosaic antigen, mesothelioma AB1-gag

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914 Theoretical Analysis of Mechanical Vibration for Offshore Platform Structures

Authors: Saeed Asiri, Yousuf Z. AL-Zahrani

Abstract:

A new class of support structures, called periodic structures, is introduced in this paper as a viable means for isolating the vibration transmitted from the sea waves to offshore platform structures through its legs. A passive approach to reduce transmitted vibration generated by waves is presented. The approach utilizes the property of periodic structural components that creates stop and pass bands. The stop band regions can be tailored to correspond to regions of the frequency spectra that contain harmonics of the wave frequency, attenuating the response in those regions. A periodic structural component is comprised of a repeating array of cells, which are themselves an assembly of elements. The elements may have differing material properties as well as geometric variations. For the purpose of this research, only geometric and material variations are considered and each cell is assumed to be identical. A periodic leg is designed in order to reduce transmitted vibration of sea waves. The effectiveness of the periodicity on the vibration levels of platform will be demonstrated theoretically. The theory governing the operation of this class of periodic structures is introduced using the transfer matrix method. The unique filtering characteristics of periodic structures are demonstrated as functions of their design parameters for structures with geometrical and material discontinuities; and determine the propagation factor by using the spectral finite element analysis and the effectiveness of design on the leg structure by changing the ratio of step length and area interface between the materials is demonstrated in order to find the propagation factor and frequency response.

Keywords: vibrations, periodic structures, offshore, platforms, transfer matrix method

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913 Towards Binder-Free and Self Supporting Flexible Supercapacitor from Carbon Nano-Onions and Their Composite with CuO Nanoparticles

Authors: Debananda Mohapatra, Subramanya Badrayyana, Smrutiranjan Parida

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Recognizing the upcoming era of carbon nanostructures and their revolutionary applications, we investigated the formation and supercapacitor application of highly pure and hydrophilic carbon nano-onions (CNOs) by economical one-step flame-synthesis procedure. The facile and scalable method uses easily available organic carbon source such as clarified butter, restricting the use of any catalyst, sophisticated instrumentation, high vacuum and post processing purification procedure. The active material was conformally coated onto a locally available cotton wipe by “sonicating and drying” process to obtain novel, lightweight, inexpensive, flexible, binder-free electrodes with strong adhesion between nanoparticles and porous wipe. This interesting electrode with CNO as the active material delivers a specific capacitance of 102.16 F/g, the energy density of 14.18 Wh/kg and power density of 2448 W/kg which are the highest values reported so far in symmetrical two electrode cell configuration with 1M Na2SO4 as an electrolyte. Incorporation of CuO nanoparticles to these functionalized CNOs by one-step hydrothermal method add up to a significant specific capacitance of 420 F/g with deliverable energy and power density at 58.33 Wh/kg and 4228 W/kg, respectively. The free standing CNOs, as well as CNO-CuO composite electrode, showed an excellent cyclic performance and stability retaining 95 and 90% initial capacitance even after 5000 charge-discharge cycles at a current density of 5 A/g. This work presents a new platform for high performance supercapacitors for next generation wearable electronic devices.

Keywords: binder-free, flame synthesis, flexible, carbon nano-onion

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912 Process Development of pVAX1/lacZ Plasmid DNA Purification Using Design of Experiment

Authors: Asavasereerat K., Teacharsripaitoon T., Tungyingyong P., Charupongrat S., Noppiboon S. Hochareon L., Kitsuban P.

Abstract:

Third generation of vaccines is based on gene therapy where DNA is introduced into patients. The antigenic or therapeutic proteins encoded from transgenes DNA triggers an immune-response to counteract various diseases. Moreover, DNA vaccine offers the customization of its ability on protection and treatment with high stability. The production of DNA vaccines become of interest. According to USFDA guidance for industry, the recommended limits for impurities from host cell are lower than 1%, and the active conformation homogeneity supercoiled DNA, is more than 80%. Thus, the purification strategy using two-steps chromatography has been established and verified for its robustness. Herein, pVax1/lacZ, a pre-approved USFDA DNA vaccine backbone, was used and transformed into E. coli strain DH5α. Three purification process parameters including sample-loading flow rate, the salt concentration in washing and eluting buffer, were studied and the experiment was designed using response surface method with central composite face-centered (CCF) as a model. The designed range of selected parameters was 10% variation from the optimized set point as a safety factor. The purity in the percentage of supercoiled conformation obtained from each chromatography step, AIEX and HIC, were analyzed by HPLC. The response data were used to establish regression model and statistically analyzed followed by Monte Carlo simulation using SAS JMP. The results on the purity of the product obtained from AIEX and HIC are between 89.4 to 92.5% and 88.3 to 100.0%, respectively. Monte Carlo simulation showed that the pVAX1/lacZ purification process is robust with confidence intervals of 0.90 in range of 90.18-91.00% and 95.88-100.00%, for AIEX and HIC respectively.

Keywords: AIEX, DNA vaccine, HIC, puification, response surface method, robustness

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911 Expression of Tissue Plasminogen Activator in Transgenic Tobacco Plants by Signal Peptides Targeting for Delivery to Apoplast, Endoplasmic Reticulum and Cytosol Spaces

Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi

Abstract:

Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA4404. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.

Keywords: tPA, recombinant, transgenic, tobacco

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910 Immunoliposomes for Co-Delivery of Doxorubicin and Ribonucleotide Reductase M2 Sirna Inhibit of Gastric Cancer Growth

Authors: Jie Gao

Abstract:

The combination of chemotherapy with gene therapy is highly effective in cancer therapy. To achieve combined therapeutic effects in human gastric cancer over expressing EGFR, we developed targeted LPD (liposome-polycation-DNA complex) conjugated with anti-EGFR (epidermal growth factor receptor) Fab’ for co-delivery of doxorubicin (DOX) and ribonucleotide reductase M2 (RRM2) siRNA (DOX-RRM2-TLPD). The results showed that EGFR was over expressed in several gastric cancer cell lines and gastric cancer tissues. Gene Expression Omnibus (GEO) results showed that RRM2 expression was significantly higher in gastric cancer than in non-gastric cancer tissue, and RRM2 siRNA inhibited the proliferation of several gastric cancer cells, indicating that RRM2 is a candidate target for gastric cancer therapy. Confocal studies and flow cytometry showed that DOX-RRM2-TLPD delivered DOX and RRM2 siRNA to EGFR over expressing gastric cancer cells specifically and efficiently both in vitro and in vivo, resulting in enhanced therapeutic effects (cytotoxicity and apoptosis) compared with single-drug loaded or non-targeted controls, including DOX-NC-TLPD (targeted LPD co-delivering DOX and negative control siRNA), RRM2-TLPD (targeted LPD delivering RRM2 siRNA) and DOX-RRM2-NTLPD (non-targeted LPD co-delivering DOX and RRM2 siRNA). The in vivo antitumor assay showed that the average weight of the gastric cancer in mice treated with DOX-RRM2-TLPD was significantly lighter than that of mice treated with other controls. DOX-RRM2-TLPD represents an effective approach for combined therapy of gastric cancer over expressing EGFR.

Keywords: gene therapy, chemotherapy, immunoliposomes, gastric cancer

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