Search results for: carragenan induced edema
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2866

Search results for: carragenan induced edema

1726 The Effect of Radiation on Unsteady MHD Flow past a Vertical Porous Plate in the Presence of Heat Flux

Authors: Pooja Sharma

Abstract:

In the present paper the effects of radiation is studied on unsteady flow of viscous incompressible electrically conducting fluid past a vertical porous plate embedded in the porous medium in the presence of constant heat flux. A uniform Transverse Magnetic field is considered and induced magnetic field is supposed as negligible. The non-linear governing equations are solved numerically. Numerical results of the velocity and temperature fields are shown through graphs. The results illustrates that the appropriator combination of regulated values of thermo-physical parameters is expedient for controlling the flow system.

Keywords: heat transfer, radiation, MHD flow, porous medium

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1725 An Investigation of Etiology of Liver Cirrhosis and Its Complications with Other Co-morbid Diseases

Authors: Tayba Akram

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our main objective of this study is to work on the etiology of liver cirrhosis, to find basic reasons and causes of liver damage, and to find the pattern of liver cirrhosis in hepatic patients either suffering from hepatitis B/C or simple jaundice. We can evaluate medical treatment and the latest trends in patients suffering from liver cirrhosis. We can evaluate the side effects and adverse effects induced by drug therapy used to treat liver cirrhosis. The conclusion is based on the etiology of liver cirrhosis. The most common cause of liver cirrhosis is the viral Hepatitis C virus. Other common causes of liver cirrhosis that are estimated from our research are Hepatitis B virus, Diabetes Mellitus, Ascites, and very rarely found Hepatitis D virus.

Keywords: etiology, liver, cirrhosis, co-morbid diseases

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1724 Aframomum melegueta Improves Antioxidant Status of Type 2 Diabetes Rats Model

Authors: Aminu Mohammed, Shahidul Islam

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Aframomum melegueta K.Schum commonly known as Grains of Paradise has been a popularly used spice in most of the African food preparation. Available data have shown that ethyl acetate fraction from crude ethanolic extract exhibited α-amylase and α-glucosidase inhibitory actions, improved pancreatic β-cell damage and ameliorated insulin resistance in diabetic rats. Additionally, 6-gingerol, 6-shogaol, 6-paradol and oleanolic acid are shown to be the compounds responsible for the antidiabetic action of A. melegueta. However, detail antioxidant potential of this spice in a diabetic animal model has not yet been reported. Thus, the present study investigates the effect of oral consumption of A. melegueta fruit on the in vivo antioxidant status of type 2 diabetes (T2D) model of rats. T2D was induced in rats by feeding a 10% fructose solution ad libitum for two weeks followed by a single intraperitoneal injection of streptozotocin (40 mg/kg body weight (bw)). The animals were orally administered with 150 (DAML) or 300 mg/kg bw (DAMH) of the fraction once daily for four weeks. Data were analyzed by using a statistical software package (SPSS for Windows, version 22, IBM Corporation, NY, USA) using Tukey’s-HSD multiple range post-hoc test. Values were considered significantly different at p < 0.05. According to the data, after four weeks of intervention, diabetic untreated animals showed significantly (p < 0.05) elevation of blood glucose levels. The levels of thiobarbituric acid reactive substances (TBARS) were observed to increase with concomitant reduction of reduced glutathione (GSH) levels in the serum and organs (liver, kidney, heart and pancreas) of diabetic untreated animals. The activities of endogenous antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, and reductase) were greatly reduced in the serum and organs of diabetic untreated animals compared to the normal animals. These alterations were reverted to near-normal after the treatment of A. melegueta fruit in the treated groups (DAML & DAMH) within the study period, especially at the dose of 300 mg/kg bw. This potent antioxidant action may partly be attributed to the presence of the 6-Gingerol, 6-shogaol and 6-paradol are known to possess antioxidant action. The results of our study showed that A. melegueta intake improved the antioxidant status of T2D rats and therefore could be used to ameliorate the diabetes-induced oxidative damage.

Keywords: Aframomum melegueta, antioxidant, ethyl acetate extract, type 2 diabetes

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1723 Comparision of Neutrophil Response to Curvularia, Bipolaris and Aspergillus Species

Authors: Eszter J. Tóth, Alexandra Hoffmann, Csaba Vágvölgyi, Tamás Papp

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Members of the genera Curvularia and Bipolaris are closely related melanin producing filamentous fungi; both of them have the teleomorph states in genus Cochliobolus. While Bipolaris species infect only plants and may cause serious agriculture damages, some Curvularia species was recovered from opportunistic human infections. The human pathogenic species typically cause phaeohyphomycoses, i.e. mould infections caused by melanised fungi, which can manifest as invasive mycoses with frequent involvement of the central nervous system in immunocompromised patients or as local infections (e.g. keratitis, sinusitis, and cutaneous lesions) in immunocompetent people. Although their plant-fungal interactions have been intensively studied, there is only little information available about the human pathogenic feature of these fungi. The aim of this study was to investigate the neutrophil granulocytes’ response to hyphal forms of Curvularia and Bipolaris in comparison with the response to Aspergillus. In the present study Curvularia lunata SZMC 23759 and Aspergillus fumigatus SZMC 23245 both isolated from human eye infection, and Bipolaris zeicola BRIP 19582b isolated from plant leaf were examined. Neutrophils were isolated from heparinised venous blood of healthy donors with dextran sedimentation followed by centrifugation over Ficoll and hypotonic lysis of erythrocytes. Viability and purity of the cells were checked with trypan blue and Wright staining, respectively. Infection of neutrophils was carried out with germinated conidia in a ratio of 5:1. Production of hydrogen peroxide, superoxide anion, and nitrogen monoxide was measured both intracellularly and extracellularly in response to the germinated spores with or without the supernatant and after serum treatment. ROS and NOS production of neutrophils in interaction with the three fungi were compared. It is already known that Aspergillus species induce ROS production of neutrophils only after serum treatment. Although, in case of C. lunata, serum opsonisation also induced an intensive production of reactive species, lower level of production was measured in the lack of serum as well. After interaction with the plant pathogenic B. zeicola, amount of reactive species found to be similar with and without serum treatment. The presence of germination supernatant decreased the reactive species production in case of each fungus. Interaction with Curvularia, Bipolaris and Aspergillus species induced different response of neutrophils. It seems that recognition of C. lunata and B. zeicola is independent of serum opsonisation, albeit it increases the level of the produced reactive species in response for C. lunata. The study was supported by the grant LP2016-8/2016.

Keywords: Curvularia, neutrophils, NOS, ROS, serum opsonisation

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1722 A Proposal of Local Indentation Techniques for Mechanical Property Evaluation

Authors: G. B. Lim, C. H. Jeon, K. H. Jung

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General light metal alloys are often developed in the material of transportation equipment such as automobiles and aircraft. Among the light metal alloys, magnesium is the lightest structural material with superior specific strength and many attractive physical and mechanical properties. However, magnesium alloys were difficult to obtain the mechanical properties at warm temperature. The aims of present work were to establish an analytical relation between mechanical properties and plastic flow induced by local indentation. An experimental investigation of the local strain distribution was carried out using a specially designed local indentation equipment in conjunction with ARAMIS based on digital image correlation method.

Keywords: indentation, magnesium, mechanical property, lightweight material, ARAMIS

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1721 Evaluation of the Analgesic Activity of Defatted Methanol Extract of Capparis spinosa L. Root Barks

Authors: Asma Meddour, Mouloud Yahia, Afaf Benhouda, Souhila Benbia, Hachani Khadhraoui

Abstract:

Peripheral analgesic activity of defatted methanol extract of root barks of Capparis spinosa was tested orally at the dose of 100 and 200 mg/kg against pain induced by acetic acid in rats. The dose of 200 mg/kg presents significant analgesic effect with a percentage of inhibition of torsions of 88.51% compared to the positive control which is the acetylsalicylic acid which represents a percentage of inhibition of 92.55%. The dose of 100 mg/kg presents a percentage of inhibition of 81.68%.

Keywords: peripheral analgesic activity, Capparis spinosa, percentage of inhibition of torsions, chemical sciences

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1720 Detecting Rat’s Kidney Inflammation Using Real Time Photoacoustic Tomography

Authors: M. Y. Lee, D. H. Shin, S. H. Park, W.C. Ham, S.K. Ko, C. G. Song

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Photoacoustic Tomography (PAT) is a promising medical imaging modality that combines optical imaging contrast with the spatial resolution of ultrasound imaging. It can also distinguish the changes in biological features. But, real-time PAT system should be confirmed due to photoacoustic effect for tissue. Thus, we have developed a real-time PAT system using a custom-developed data acquisition board and ultrasound linear probe. To evaluate performance of our system, phantom test was performed. As a result of those experiments, the system showed satisfactory performance and its usefulness has been confirmed. We monitored the degradation of inflammation which induced on the rat’s kidney using real-time PAT.

Keywords: photoacoustic tomography, inflammation detection, rat, kidney, contrast agent, ultrasound

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1719 Synthesis and Anticholinesterase Activity of Carvacrol Derivatives

Authors: Fatih Sonmez

Abstract:

Alzheimer’s disease (AD) is a progressive neurodegenerative disease and it is the most common form of dementia that affects aged people. Acetylcholinesterase is a hydrolase involved in the termination of impulse transmission at cholinergic synapses by rapid hydrolysis of the neurotransmitter ACh in the central and peripheral nervous system. Carvacrol (5-iso-propyl-2-methyl-phenol) is a main bioactive monoterpene isolated from many medicinal herbs, such as Thymus vulgaris, Monarda punctate and Origanum vulgare spp. It is known that carvacrol has been widely used as an active anti-inflammatory ingredient, which can inhibit the isoproterenol induced inflammation in myocardial infarcted rats. In this paper, a series of 12 carvacrol substituted carbamate derivatives (2a-l) was synthesized and their inhibitory activities on AChE and BuChE were evaluated. Among them, 2d exhibited the strongest inhibition against AChE with an IC50 value of 2.22 µM, which was 130-fold more than that of carvacrol (IC50 = 288.26 µM).

Keywords: Acetylcholinesterase, Butyrylcholinesterase, Carbamate, Carvacrol

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1718 Data-Driven Surrogate Models for Damage Prediction of Steel Liquid Storage Tanks under Seismic Hazard

Authors: Laura Micheli, Majd Hijazi, Mahmoud Faytarouni

Abstract:

The damage reported by oil and gas industrial facilities revealed the utmost vulnerability of steel liquid storage tanks to seismic events. The failure of steel storage tanks may yield devastating and long-lasting consequences on built and natural environments, including the release of hazardous substances, uncontrolled fires, and soil contamination with hazardous materials. It is, therefore, fundamental to reliably predict the damage that steel liquid storage tanks will likely experience under future seismic hazard events. The seismic performance of steel liquid storage tanks is usually assessed using vulnerability curves obtained from the numerical simulation of a tank under different hazard scenarios. However, the computational demand of high-fidelity numerical simulation models, such as finite element models, makes the vulnerability assessment of liquid storage tanks time-consuming and often impractical. As a solution, this paper presents a surrogate model-based strategy for predicting seismic-induced damage in steel liquid storage tanks. In the proposed strategy, the surrogate model is leveraged to reduce the computational demand of time-consuming numerical simulations. To create the data set for training the surrogate model, field damage data from past earthquakes reconnaissance surveys and reports are collected. Features representative of steel liquid storage tank characteristics (e.g., diameter, height, liquid level, yielding stress) and seismic excitation parameters (e.g., peak ground acceleration, magnitude) are extracted from the field damage data. The collected data are then utilized to train a surrogate model that maps the relationship between tank characteristics, seismic hazard parameters, and seismic-induced damage via a data-driven surrogate model. Different types of surrogate algorithms, including naïve Bayes, k-nearest neighbors, decision tree, and random forest, are investigated, and results in terms of accuracy are reported. The model that yields the most accurate predictions is employed to predict future damage as a function of tank characteristics and seismic hazard intensity level. Results show that the proposed approach can be used to estimate the extent of damage in steel liquid storage tanks, where the use of data-driven surrogates represents a viable alternative to computationally expensive numerical simulation models.

Keywords: damage prediction , data-driven model, seismic performance, steel liquid storage tanks, surrogate model

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1717 Stress Study in Implants Dental

Authors: M. Benlebna, B. Serier, B. Bachir Bouiadjra, S. Khalkhal

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This study focuses on the mechanical behavior of a dental prosthesis subjected to dynamic loads chewing. It covers a three-dimensional analysis by the finite element method, the level of distribution of equivalent stresses induced in the bone between the implants (depending on the number of implants). The studied structure, consisting of a braced, implant and mandibular bone is subjected to dynamic loading of variable amplitude in three directions corrono-apical, mesial-distal and bucco-lingual. These efforts simulate those of mastication. We show that compared to the implantation of a single implant, implantology using two implants promotes the weakening of the bones. This weakness is all the more likely that the implants are located in close proximity to one another.

Keywords: stress, bone, dental implant, distribution, stress levels, dynamic, effort, interaction, prosthesis

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1716 Computational Analysis of Thermal Degradation in Wind Turbine Spars' Equipotential Bonding Subjected to Lightning Strikes

Authors: Antonio A. M. Laudani, Igor O. Golosnoy, Ole T. Thomsen

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Rotor blades of large, modern wind turbines are highly susceptible to downward lightning strikes, as well as to triggering upward lightning; consequently, it is necessary to equip them with an effective lightning protection system (LPS) in order to avoid any damage. The performance of existing LPSs is affected by carbon fibre reinforced polymer (CFRP) structures, which lead to lightning-induced damage in the blades, e.g. via electrical sparks. A solution to prevent internal arcing would be to electrically bond the LPS and the composite structures such that to obtain the same electric potential. Nevertheless, elevated temperatures are achieved at the joint interfaces because of high contact resistance, which melts and vaporises some of the epoxy resin matrix around the bonding. The produced high-pressure gasses open up the bonding and can ignite thermal sparks. The objective of this paper is to predict the current density distribution and the temperature field in the adhesive joint cross-section, in order to check whether the resin pyrolysis temperature is achieved and any damage is expected. The finite element method has been employed to solve both the current and heat transfer problems, which are considered weakly coupled. The mathematical model for electric current includes Maxwell-Ampere equation for induced electric field solved together with current conservation, while the thermal field is found from heat diffusion equation. In this way, the current sub-model calculates Joule heat release for a chosen bonding configuration, whereas the thermal analysis allows to determining threshold values of voltage and current density not to be exceeded in order to maintain the temperature across the joint below the pyrolysis temperature, therefore preventing the occurrence of outgassing. In addition, it provides an indication of the minimal number of bonding points. It is worth to mention that the numerical procedures presented in this study can be tailored and applied to any type of joints other than adhesive ones for wind turbine blades. For instance, they can be applied for lightning protection of aerospace bolted joints. Furthermore, they can even be customized to predict the electromagnetic response under lightning strikes of other wind turbine systems, such as nacelle and hub components.

Keywords: carbon fibre reinforced polymer, equipotential bonding, finite element method, FEM, lightning protection system, LPS, wind turbine blades

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1715 Radiation Induced DNA Damage and Its Modification by Herbal Preparation of Hippophae rhamnoides L. (SBL-1): An in vitro and in vivo Study in Mice

Authors: Anuranjani Kumar, Madhu Bala

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Ionising radiation exposure induces generation of free radicals and the oxidative DNA damage. SBL-1, a radioprotective leaf extract prepared from leaves Hippophae rhamnoides L. (Common name; Seabuckthorn), showed > 90% survival in mice population that was treated with lethal dose (10 Gy) of ⁶⁰Co gamma irradiation. In this study, early effects of pre-treatment with or without SBL-1 in blood peripheral blood lymphocytes (PBMCs) were investigated by cell viability assays (trypan blue and MTT). The quantitative in vitro study of Hoescht/PI staining was performed to check the apoptosis/necrosis in PBMCs irradiated at 2 Gy with or without pretreatment of SBL-1 (at different concentrations) up to 24 and 48h. Comet assay was performed in vivo, to detect the DNA strands breaks and its repair mechanism on peripheral blood lymphocytes at lethal dose (10 Gy). For this study, male mice (wt. 28 ± 2g) were administered radioprotective dose (30mg/kg body weight) of SBL-1, 30 min prior to irradiation. Animals were sacrificed at 24h and 48h. Blood was drawn through cardiac puncture, and blood lymphocytes were separated using histopaque column. Both neutral and alkaline comet assay were performed using standardized technique. In irradiated animals, alkaline comet assay revealed single strand breaks (SSBs) that showed significant (p < 0.05) increase in percent DNA in tail and Olive tail moment (OTM) at 24 h while at 48h the percent DNA in tail further increased significantly (p < 0.02). The double strands breaks (DSBs) increased significantly (p < 0.01) at 48 h in neutral assay, in comparison to untreated control. The animals pre-treated with SBL-1 before irradiation showed significantly (p < 0.05) less DSBs at 48 h treatment in comparison to irradiated group of animals. The SBL-1 alone treated group itself showed no toxicity. The antioxidant potential of SBL-1 were also investigated by in vitro biochemical assays such as DPPH (p < 0.05), ABTS, reducing ability (p < 0.09), hydroxyl radical scavenging (p < 0.05), ferric reducing antioxidant power (FRAP), superoxide radical scavenging activity (p < 0.05), hydrogen peroxide scavenging activity (p < 0.05) etc. SBL-1 showed strong free radical scavenging power that plays important role in the studies of radiation-induced injuries. The SBL-1 treated PBMCs showed significant (p < 0.02) viability in trypan blue assay at 24-hour incubation.

Keywords: radiation, SBL-1, SSBs, DSBs, FRAP, PBMCs

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1714 Development of Agomelatine Loaded Proliposomal Powders for Improved Intestinal Permeation: Effect of Surface Charge

Authors: Rajasekhar Reddy Poonuru, Anusha Parnem

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Purpose: To formulate proliposome powder of agomelatine, an antipsychotic drug, and to evaluate physicochemical, in vitro characters and effect of surface charge on ex vivo intestinal permeation. Methods: Film deposition technique was employed to develop proliposomal powders of agomelatin with varying molar ratios of lipid Hydro Soy PC L-α-phosphatidylcholine (HSPC) and cholesterol with fixed sum of drug. With the aim to derive free flowing and stable proliposome powder, fluid retention potential of various carriers was examined. Liposome formation and number of vesicles formed for per mm3 up on hydration, vesicle size, and entrapment efficiency was assessed to deduce an optimized formulation. Sodium cholate added to optimized formulation to induce surface charge on formed vesicles. Solid-state characterization (FTIR, DSC, and XRD) was performed with the intention to assess native crystalline and chemical behavior of drug. The in vitro dissolution test of optimized formulation along with pure drug was evaluated to estimate dissolution efficiency (DE) and relative dissolution rate (RDR). Effective permeability co-efficient (Peff(rat)) in rat and enhancement ratio (ER) of drug from formulation and pure drug dispersion were calculated from ex vivo permeation studies in rat ileum. Results: Proliposomal powder formulated with equimolar ratio of HSPC and cholesterol ensued in higher no. of vesicles (3.95) with 90% drug entrapment up on hydration. Neusilin UFL2 was elected as carrier because of its high fluid retention potential (4.5) and good flow properties. Proliposome powder exhibited augmentation in DE (60.3 ±3.34) and RDR (21.2±01.02) of agomelation over pure drug. Solid state characterization studies demonstrated the transformation of native crystalline form of drug to amorphous and/or molecular state, which was in correlation with results obtained from in vitro dissolution test. The elevated Peff(rat) of 46.5×10-4 cm/sec and ER of 2.65 of drug from charge induced proliposome formulation with respect to pure drug dispersion was assessed from ex vivo intestinal permeation studies executed in ileum of wistar rats. Conclusion: Improved physicochemical characters and ex vivo intestinal permeation of drug from charge induced proliposome powder with Neusilin UFL2 unravels the potentiality of this system in enhancing oral delivery of agomelatin.

Keywords: agomelatin, proliposome, sodium cholate, neusilin

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1713 Analyzing Bridge Response to Wind Loads and Optimizing Design for Wind Resistance and Stability

Authors: Abdul Haq

Abstract:

The goal of this research is to better understand how wind loads affect bridges and develop strategies for designing bridges that are more stable and resistant to wind. The effect of wind on bridges is essential to their safety and functionality, especially in areas that are prone to high wind speeds or violent wind conditions. The study looks at the aerodynamic forces and vibrations caused by wind and how they affect bridge construction. Part of the research method involves first understanding the underlying ideas influencing wind flow near bridges. Computational fluid dynamics (CFD) simulations are used to model and forecast the aerodynamic behaviour of bridges under different wind conditions. These models incorporate several factors, such as wind directionality, wind speed, turbulence intensity, and the influence of nearby structures or topography. The results provide significant new insights into the loads and pressures that wind places on different bridge elements, such as decks, pylons, and connections. Following the determination of the wind loads, the structural response of bridges is assessed. By simulating their dynamic behavior under wind-induced forces, Finite Element Analysis (FEA) is used to model the bridge's component parts. This work contributes to the understanding of which areas are at risk of experiencing excessive stresses, vibrations, or oscillations due to wind excitations. Because the bridge has inherent modes and frequencies, the study considers both static and dynamic responses. Various strategies are examined to maximize the design of bridges to withstand wind. It is possible to alter the bridge's geometry, add aerodynamic components, add dampers or tuned mass dampers to lessen vibrations, and boost structural rigidity. Through an analysis of several design modifications and their effectiveness, the study aims to offer guidelines and recommendations for wind-resistant bridge design. In addition to the numerical simulations and analyses, there are experimental studies. In order to assess the computational models and validate the practicality of proposed design strategies, scaled bridge models are tested in a wind tunnel. These investigations help to improve numerical models and prediction precision by providing valuable information on wind-induced forces, pressures, and flow patterns. Using a combination of numerical models, actual testing, and long-term performance evaluation, the project aims to offer practical insights and recommendations for building wind-resistant bridges that are secure, long-lasting, and comfortable for users.

Keywords: wind effects, aerodynamic forces, computational fluid dynamics, finite element analysis

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1712 Failure Localization of Bipolar Integrated Circuits by Implementing Active Voltage Contrast

Authors: Yiqiang Ni, Xuanlong Chen, Enliang Li, Linting Zheng, Shizheng Yang

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Bipolar ICs are playing an important role in military applications, mainly used in logic gates, such as inverter and NAND gate. The defect of metal break located on the step is one of the main failure mechanisms of bipolar ICs, resulting in open-circuit or functional failure. In this situation, general failure localization methods like optical beam-induced resistance change (OBIRCH) and photon emission microscopy (PEM) might not be fully effective. However, active voltage contrast (AVC) can be used as a voltage probe, which may pinpoint the incorrect potential and thus locate the failure position. Two case studies will be present in this paper on how to implement AVC for failure localization, and the detailed failure mechanism will be discussed.

Keywords: bipolar IC, failure localization, metal break, open failure, voltage contrast

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1711 Mathematical Modeling of Cell Volume Alterations under Different Osmotic Conditions

Authors: Juliana A. Knocikova, Yann Bouret, Médéric Argentina, Laurent Counillon

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Cell volume, together with membrane potential and intracellular hydrogen ion concentration, is an essential biophysical parameter for normal cellular activity. Cell volumes can be altered by osmotically active compounds and extracellular tonicity. In this study, a simple mathematical model of osmotically induced cell swelling and shrinking is presented. Emphasis is given to water diffusion across the membrane. The mathematical description of the cellular behavior consists in a system of coupled ordinary differential equations. We compare experimental data of cell volume alterations driven by differences in osmotic pressure with mathematical simulations under hypotonic and hypertonic conditions. Implications for a future model are also discussed.

Keywords: eukaryotic cell, mathematical modeling, osmosis, volume alterations

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1710 Thermoplastic Composites with Reduced Discoloration and Enhanced Fire-Retardant Property

Authors: Peng Cheng, Liqing Wei, Hongyu Chen, Ruomiao Wang

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This paper discusses a light-weight reinforced thermoplastic (LWRT) composite with superior fire retardancy. This porous LWRT composite is manufactured using polyolefin, fiberglass, and fire retardant additives via a wet-lay process. However, discoloration of the LWRT can be induced by various mechanisms, which may be a concern in the building and construction industry. It is commonly understood that discoloration is strongly associated with the presence of phenolic antioxidant(s) and NOx. The over-oxidation of phenolic antioxidant(s) is probably the root-cause of the discoloration (pinking/yellowing). Hanwha Azdel, Inc. developed a LWRT with fire-retardant property of ASTM E84-Class A specification, as well as negligible discoloration even under harsh conditions. In addition, this thermoplastic material is suitable for secondary processing (e.g. compression molding) if necessary.

Keywords: discoloration, fire-retardant, thermoplastic composites, wet-lay process

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1709 In Vivo Evaluation of Exposure to Electromagnetic Fields at 27 GHz (5G) of Danio Rerio: A Preliminary Study

Authors: Elena Maria Scalisi, Roberta Pecoraro, Martina Contino, Sara Ignoto, Carmelo Iaria, Santi Concetto Pavone, Gino Sorbello, Loreto Di Donato, Maria Violetta Brundo

Abstract:

5G Technology is evolving to satisfy a variety of service requirements that may allow high data-rate connections (1Gbps) and lower latency times than current (<1ms). In order to support a high data transmission speed and a high traffic service for eMBB (enhanced mobile broadband) use cases, 5G systems have the characteristic of using different frequency bands of the radio wave spectrum (700 MHz, 3.6-3.8 GHz and 26.5-27.5 GHz), thus taking advantage of higher frequencies than previous mobile radio generations (1G-4G). However, waves at higher frequencies have a lower capacity to propagate in free space and therefore, in order to guarantee the capillary coverage of the territory for high reliability applications, it will be necessary to install a large number of repeaters. Following the introduction of this new technology, there has been growing concern over the past few months about possible harmful effects on human health. The aim of this preliminary study is to evaluate possible short term effects induced by 5G-millimeter waves on embryonic development and early life stages of Danio rerio by Z-FET. We exposed developing zebrafish at frequency of 27 GHz, with a standard pyramidal horn antenna placed at 15 cm far from the samples holder ensuring an incident power density of 10 mW/cm2. During the exposure cycle, from 6 h post fertilization (hpf) to 96 hpf, we measured a different morphological endpoints every 24 hours. Zebrafish embryo toxicity test (Z-FET) is a short term test, carried out on fertilized eggs of zebrafish and it represents an effective alternative to acute test with adult fish (OECD, 2013). We have observed that 5G did not reveal significant impacts on mortality nor on morphology because exposed larvae showed a normal detachment of the tail, presence of heartbeat, well-organized somites, therefore hatching rate was lower than untreated larvae even at 48 h of exposure. Moreover, the immunohistochemical analysis performed on larvae showed a negativity to the HSP-70 expression used as a biomarkers. This is a preliminary study on evaluation of potential toxicity induced by 5G and it seems appropriate to underline the importance that further studies would take, aimed at clarifying the probable real risk of exposure to electromagnetic fields.

Keywords: Biomarker of exposure, embryonic development, 5G waves, zebrafish embryo toxicity test

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1708 Raman Tweezers Spectroscopy Study of Size Dependent Silver Nanoparticles Toxicity on Erythrocytes

Authors: Surekha Barkur, Aseefhali Bankapur, Santhosh Chidangil

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Raman Tweezers technique has become prevalent in single cell studies. This technique combines Raman spectroscopy which gives information about molecular vibrations, with optical tweezers which use a tightly focused laser beam for trapping the single cells. Thus Raman Tweezers enabled researchers analyze single cells and explore different applications. The applications of Raman Tweezers include studying blood cells, monitoring blood-related disorders, silver nanoparticle-induced stress, etc. There is increased interest in the toxic effect of nanoparticles with an increase in the various applications of nanoparticles. The interaction of these nanoparticles with the cells may vary with their size. We have studied the effect of silver nanoparticles of sizes 10nm, 40nm, and 100nm on erythrocytes using Raman Tweezers technique. Our aim was to investigate the size dependence of the nanoparticle effect on RBCs. We used 785nm laser (Starbright Diode Laser, Torsana Laser Tech, Denmark) for both trapping and Raman spectroscopic studies. 100 x oil immersion objectives with high numerical aperture (NA 1.3) is used to focus the laser beam into a sample cell. The back-scattered light is collected using the same microscope objective and focused into the spectrometer (Horiba Jobin Vyon iHR320 with 1200grooves/mm grating blazed at 750nm). Liquid nitrogen cooled CCD (Symphony CCD-1024x256-OPEN-1LS) was used for signal detection. Blood was drawn from healthy volunteers in vacutainer tubes and centrifuged to separate the blood components. 1.5 ml of silver nanoparticles was washed twice with distilled water leaving 0.1 ml silver nanoparticles in the bottom of the vial. The concentration of silver nanoparticles is 0.02mg/ml so the 0.03mg of nanoparticles will be present in the 0.1 ml nanoparticles obtained. The 25 ul of RBCs were diluted in 2 ml of PBS solution and then treated with 50 ul (0.015mg) of nanoparticles and incubated in CO2 incubator. Raman spectroscopic measurements were done after 24 hours and 48 hours of incubation. All the spectra were recorded with 10mW laser power (785nm diode laser), 60s of accumulation time and 2 accumulations. Major changes were observed in the peaks 565 cm-1, 1211 cm-1, 1224 cm-1, 1371 cm-1, 1638 cm-1. A decrease in intensity of 565 cm-1, increase in 1211 cm-1 with a reduction in 1224 cm-1, increase in intensity of 1371 cm-1 also peak disappearing at 1635 cm-1 indicates deoxygenation of hemoglobin. Nanoparticles with higher size were showing maximum spectral changes. Lesser changes observed in case of 10nm nanoparticle-treated erythrocyte spectra.

Keywords: erythrocytes, nanoparticle-induced toxicity, Raman tweezers, silver nanoparticles

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1707 Influence and Interaction of Temperature, H2S and pH on Concrete Sewer Pipe Corrosion

Authors: Anna Romanova, Mojtaba Mahmoodian, Morteza A. Alani

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Concrete sewer pipes are known to suffer from a process of hydrogen sulfide gas induced sulfuric acid corrosion. This leads to premature pipe degradation, performance failure and collapses which in turn may lead to property and health damage. The above work reports on a field study undertaken in working sewer manholes where the parameters of effluent temperature and pH as well as ambient temperature and concentration of hydrogen sulfide were continuously measured over a period of two months. Early results suggest that effluent pH has no direct effect on hydrogen sulfide build up; on average the effluent temperature is 3.5°C greater than the ambient temperature inside the manhole and also it was observed that hydrogen sulfate concentration increases with increasing temperature.

Keywords: concrete corrosion, hydrogen sulfide gas, temperature, sewer pipe

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1706 High-Pressure Polymorphism of 4,4-Bipyridine Hydrobromide

Authors: Michalina Aniola, Andrzej Katrusiak

Abstract:

4,4-Bipyridine is an important compound often used in chemical practice and more recently frequently applied for designing new metal organic framework (MoFs). Here we present a systematic high-pressure study of its hydrobromide salt. 4,4-Bipyridine hydrobromide monohydrate, 44biPyHBrH₂O, at ambient-pressure is orthorhombic, space group P212121 (phase a). Its hydrostatic compression shows that it is stable to 1.32 GPa at least. However, the recrystallization above 0.55 GPa reveals a new hidden b-phase (monoclinic, P21/c). Moreover, when the 44biPyHBrH2O is heated to high temperature the chemical reactions of this compound in methanol solution can be observed. High-pressure experiments were performed using a Merrill-Bassett diamond-anvil cell (DAC), modified by mounting the anvils directly on the steel supports, and X-ray diffraction measurements were carried out on a KUMA and Excalibur diffractometer equipped with an EOS CCD detector. At elevated pressure, the crystal of 44biPyHBrH₂O exhibits several striking and unexpected features. No signs of instability of phase a were detected to 1.32 GPa, while phase b becomes stable at above 0.55 GPa, as evidenced by its recrystallizations. Phases a and b of 44biPyHBrH2O are partly isostructural: their unit-cell dimensions and the arrangement of ions and water molecules are similar. In phase b the HOH-Br- chains double the frequency of their zigzag motifs, compared to phase a, and the 44biPyH+ cations change their conformation. Like in all monosalts of 44biPy determined so far, in phase a the pyridine rings are twisted by about 30 degrees about bond C4-C4 and in phase b they assume energy-unfavorable planar conformation. Another unusual feature of 44biPyHBrH2O is that all unit-cell parameters become longer on the transition from phase a to phase b. Thus the volume drop on the transition to high-pressure phase b totally depends on the shear strain of the lattice. Higher temperature triggers chemical reactions of 44biPyHBrH2O with methanol. When the saturated methanol solution compound precipitated at 0.1 GPa and temperature of 423 K was required to dissolve all the sample, the subsequent slow recrystallization at isochoric conditions resulted in disalt 4,4-bipyridinium dibromide. For the 44biPyHBrH2O sample sealed in the DAC at 0.35 GPa, then dissolved at isochoric conditions at 473 K and recrystallized by slow controlled cooling, a reaction of N,N-dimethylation took place. It is characteristic that in both high-pressure reactions of 44biPyHBrH₂O the unsolvated disalt products were formed and that free base 44biPy and H₂O remained in the solution. The observed reactions indicate that high pressure destabilized ambient-pressure salts and favors new products. Further studies on pressure-induced reactions are carried out in order to better understand the structural preferences induced by pressure.

Keywords: conformation, high-pressure, negative area compressibility, polymorphism

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1705 The Association between Prior Antibiotic Use and Subsequent Risk of Infectious Disease: A Systematic Review

Authors: Umer Malik, David Armstrong, Mark Ashworth, Alex Dregan, Veline L'Esperance, Lucy McDonnell, Mariam Molokhia, Patrick White

Abstract:

Introduction: The microbiota lining epithelial surfaces is thought to play an important role in many human physiological functions including defense against pathogens and modulation of immune response. The microbiota is susceptible to disruption from external influences such as exposure to antibiotic medication. It is thought that antibiotic-induced disruption of the microbiota could predispose to pathogen overgrowth and invasion. We hypothesized that antibiotic use would be associated with increased risk of future infections. We carried out a systematic review of evidence of associations between antibiotic use and subsequent risk of community-acquired infections. Methods: We conducted a review of the literature for observational studies assessing the association between antibiotic use and subsequent community-acquired infection. Eligible studies were published before April 29th, 2016. We searched MEDLINE, EMBASE, and Web of Science and screened titles and abstracts using a predefined search strategy. Infections caused by Clostridium difficile, drug-resistant organisms and fungal organisms were excluded as their association with prior antibiotic use has been examined in previous systematic reviews. Results: Eighteen out of 21,518 retrieved studies met the inclusion criteria. The association between past antibiotic exposure and subsequent increased risk of infection was reported in 16 studies, including one study on Campylobacter jejuni infection (Odds Ratio [OR] 3.3), two on typhoid fever (ORs 5.7 and 12.2), one on Staphylococcus aureus skin infection (OR 2.9), one on invasive pneumococcal disease (OR 1.57), one on recurrent furunculosis (OR 16.6), one on recurrent boils and abscesses (Risk ratio 1.4), one on upper respiratory tract infection (OR 2.3) and urinary tract infection (OR 1.1), one on invasive Haemophilus influenzae type b (Hib) infection (OR 1.51), one on infectious mastitis (OR 5.38), one on meningitis (OR 2.04) and five on Salmonella enteric infection (ORs 1.4, 1.59, 1.9, 2.3 and 3.8). The effect size in three studies on Salmonella enteric infection was of marginal statistical significance. A further two studies on Salmonella infection did not demonstrate a statistically significant association between prior antibiotic exposure and subsequent infection. Conclusion: We have found an association between past antibiotic exposure and subsequent risk of a diverse range of infections in the community setting. Our findings provide evidence to support the hypothesis that prior antibiotic usage may predispose to future infection risk, possibly through antibiotic-induced alteration of the microbiota. The findings add further weight to calls to minimize inappropriate antibiotic prescriptions.

Keywords: antibiotic, infection, risk factor, side effect

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1704 Lessons from Patients Expired due to Severe Head Injuries Treated in Intensive Care Unit of Lady Reading Hospital Peshawar

Authors: Mumtaz Ali, Hamzullah Khan, Khalid Khanzada, Shahid Ayub, Aurangzeb Wazir

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Objective: To analyse the death of patients treated in neuro-surgical ICU for severe head injuries from different perspectives. The evaluation of the data so obtained to help improve the health care delivery to this group of patients in ICU. Study Design: It is a descriptive study based on retrospective analysis of patients presenting to neuro-surgical ICU in Lady Reading Hospital, Peshawar. Study Duration: It covered the period between 1st January 2009 to 31st December 2009. Material and Methods: The Clinical record of all the patients presenting with the clinical radiological and surgical features of severe head injuries, who expired in neuro-surgical ICU was collected. A separate proforma which mentioned age, sex, time of arrival and death, causes of head injuries, the radiological features, the clinical parameters, the surgical and non surgical treatment given was used. The average duration of stay and the demographic and domiciliary representation of these patients was noted. The record was analyzed accordingly for discussion and recommendations. Results: Out of the total 112 (n-112) patients who expired in one year in the neuro-surgical ICU the young adults made up the majority 64 (57.14%) followed by children, 34 (30.35%) and then the elderly age group: 10 (8.92%). Road traffic accidents were the major cause of presentation, 75 (66.96%) followed by history of fall; 23 (20.53%) and then the fire arm injuries; 13 (11.60%). The predominant CT scan features of these patients on presentation was cerebral edema, and midline shift (diffuse neuronal injuries). 46 (41.07%) followed by cerebral contusions. 28 (25%). The correctable surgical causes were present only in 18 patients (16.07%) and the majority 94 (83.92%) were given conservative management. Of the 69 (n=69) patients in which CT scan was repeated; 62 (89.85%) showed worsening of the initial CT scan abnormalities while in 7 cases (10.14%) the features were static. Among the non surgical cases both ventilatory therapy in 7 (6.25%) and tracheostomy in 39 (34.82%) failed to change the outcome. The maximum stay in the neuro ICU leading upto the death was 48 hours in 35 (31.25%) cases followed by 31 (27.67%) cases in 24 hours; 24 (21.42%) in one week and 16 (14.28%) in 72 hours. Only 6 (5.35%) patients survived more than a week. Patients were received from almost all the districts of NWFP except. The Hazara division. There were some Afghan refugees as well. Conclusion: Mortality following the head injuries is alarmingly high despite repeated claims about the professional and administrative improvement. Even places like ICU could not change the out come according to the desired aims and objectives in the present set up. A rethinking is needed both at the individual and institutional level among the concerned quarters with a clear aim at the more scientific grounds. Only then one can achieve the desired results.

Keywords: Glasgow Coma Scale, pediatrics, geriatrics, Peshawar

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1703 Lattice Twinning and Detwinning Processes in Phase Transformation in Shape Memory Alloys

Authors: Osman Adiguzel

Abstract:

Shape memory effect is a peculiar property exhibited by certain alloy systems and based on martensitic transformation, and shape memory properties are closely related to the microstructures of the material. Shape memory effect is linked with martensitic transformation, which is a solid state phase transformation and occurs with the cooperative movement of atoms by means of lattice invariant shears on cooling from high-temperature parent phase. Lattice twinning and detwinning can be considered as elementary processes activated during the transformation. Thermally induced martensite occurs as martensite variants, in self-accommodating manner and consists of lattice twins. Also, this martensite is called the twinned martensite or multivariant martensite. Deformation of shape memory alloys in martensitic state proceeds through a martensite variant reorientation. The martensite variants turn into the reoriented single variants with deformation, and the reorientation process has great importance for the shape memory behavior. Copper based alloys exhibit this property in metastable β- phase region, which has DO3 –type ordered lattice in ternary case at high temperature, and these structures martensiticaly turn into the layered complex structures with lattice twinning mechanism, on cooling from high temperature parent phase region. The twinning occurs as martensite variants with lattice invariant shears in two opposite directions, <110 > -type directions on the {110}- type plane of austenite matrix. Lattice invariant shear is not uniform in copper based ternary alloys and gives rise to the formation of unusual layered structures, like 3R, 9R, or 18R depending on the stacking sequences on the close-packed planes of the ordered lattice. The unit cell and periodicity are completed through 18 atomic layers in case of 18R-structure. On the other hand, the deformed material recovers the original shape on heating above the austenite finish temperature. Meanwhile, the material returns to the twinned martensite structures (thermally induced martensite structure) in one way (irreversible) shape memory effect on cooling below the martensite finish temperature, whereas the material returns to the detwinned martensite structure (deformed martensite) in two-way (reversible) shape memory effect. Shortly one can say that the microstructural mechanisms, responsible for the shape memory effect are the twinning and detwinning processes as well as martensitic transformation. In the present contribution, x-ray diffraction, transmission electron microscopy (TEM) and differential scanning calorimetry (DSC) studies were carried out on two copper-based ternary alloys, CuZnAl, and CuAlMn.

Keywords: shape memory effect, martensitic transformation, twinning and detwinning, layered structures

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1702 Quantification of Site Nonlinearity Based on HHT Analysis of Seismic Recordings

Authors: Ruichong Zhang

Abstract:

This study proposes a recording-based approach to characterize and quantify earthquake-induced site nonlinearity, exemplified as soil nonlinearity and/or liquefaction. Alternative to Fourier spectral analysis (FSA), the paper introduces time-frequency analysis of earthquake ground motion recordings with the aid of so-called Hilbert-Huang transform (HHT), and offers justification for the HHT in addressing the nonlinear features shown in the recordings. With the use of the 2001 Nisqually earthquake recordings, this study shows that the proposed approach is effective in characterizing site nonlinearity and quantifying the influences in seismic ground responses.

Keywords: site nonlinearity, site amplification, site damping, Hilbert-Huang Transform (HHT), liquefaction, 2001 Nisqually Earthquake

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1701 Prediction of Time to Crack Reinforced Concrete by Chloride Induced Corrosion

Authors: Anuruddha Jayasuriya, Thanakorn Pheeraphan

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In this paper, a review of different mathematical models which can be used as prediction tools to assess the time to crack reinforced concrete (RC) due to corrosion is investigated. This investigation leads to an experimental study to validate a selected prediction model. Most of these mathematical models depend upon the mechanical behaviors, chemical behaviors, electrochemical behaviors or geometric aspects of the RC members during a corrosion process. The experimental program is designed to verify the accuracy of a well-selected mathematical model from a rigorous literature study. Fundamentally, the experimental program exemplifies both one-dimensional chloride diffusion using RC squared slab elements of 500 mm by 500 mm and two-dimensional chloride diffusion using RC squared column elements of 225 mm by 225 mm by 500 mm. Each set consists of three water-to-cement ratios (w/c); 0.4, 0.5, 0.6 and two cover depths; 25 mm and 50 mm. 12 mm bars are used for column elements and 16 mm bars are used for slab elements. All the samples are subjected to accelerated chloride corrosion in a chloride bath of 5% (w/w) sodium chloride (NaCl) solution. Based on a pre-screening of different models, it is clear that the well-selected mathematical model had included mechanical properties, chemical and electrochemical properties, nature of corrosion whether it is accelerated or natural, and the amount of porous area that rust products can accommodate before exerting expansive pressure on the surrounding concrete. The experimental results have shown that the selected model for both one-dimensional and two-dimensional chloride diffusion had ±20% and ±10% respective accuracies compared to the experimental output. The half-cell potential readings are also used to see the corrosion probability, and experimental results have shown that the mass loss is proportional to the negative half-cell potential readings that are obtained. Additionally, a statistical analysis is carried out in order to determine the most influential factor that affects the time to corrode the reinforcement in the concrete due to chloride diffusion. The factors considered for this analysis are w/c, bar diameter, and cover depth. The analysis is accomplished by using Minitab statistical software, and it showed that cover depth is the significant effect on the time to crack the concrete from chloride induced corrosion than other factors considered. Thus, the time predictions can be illustrated through the selected mathematical model as it covers a wide range of factors affecting the corrosion process, and it can be used to predetermine the durability concern of RC structures that are vulnerable to chloride exposure. And eventually, it is further concluded that cover thickness plays a vital role in durability in terms of chloride diffusion.

Keywords: accelerated corrosion, chloride diffusion, corrosion cracks, passivation layer, reinforcement corrosion

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1700 Stability Analysis of Two-delay Differential Equation for Parkinson's Disease Models with Positive Feedback

Authors: M. A. Sohaly, M. A. Elfouly

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Parkinson's disease (PD) is a heterogeneous movement disorder that often appears in the elderly. PD is induced by a loss of dopamine secretion. Some drugs increase the secretion of dopamine. In this paper, we will simply study the stability of PD models as a nonlinear delay differential equation. After a period of taking drugs, these act as positive feedback and increase the tremors of patients, and then, the differential equation has positive coefficients and the system is unstable under these conditions. We will present a set of suggested modifications to make the system more compatible with the biodynamic system. When giving a set of numerical examples, this research paper is concerned with the mathematical analysis, and no clinical data have been used.

Keywords: Parkinson's disease, stability, simulation, two delay differential equation

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1699 The Beneficial Effects of Inhibition of Hepatic Adaptor Protein Phosphotyrosine Interacting with PH Domain and Leucine Zipper 2 on Glucose and Cholesterol Homeostasis

Authors: Xi Chen, King-Yip Cheng

Abstract:

Hypercholesterolemia, characterized by high low-density lipoprotein cholesterol (LDL-C), raises cardiovascular events in patients with type 2 diabetes (T2D). Although several drugs, such as statin and PCSK9 inhibitors, are available for the treatment of hypercholesterolemia, they exert detrimental effects on glucose metabolism and hence increase the risk of T2D. On the other hand, the drugs used to treat T2D have minimal effect on improving the lipid profile. Therefore, there is an urgent need to develop treatments that can simultaneously improve glucose and lipid homeostasis. Adaptor protein phosphotyrosine interacting with PH domain and leucine zipper 2 (APPL2) causes insulin resistance in the liver and skeletal muscle via inhibiting insulin and adiponectin actions in animal models. Single-nucleotide polymorphisms in the APPL2 gene were associated with LDL-C, non-alcoholic fatty liver disease, and coronary artery disease in humans. The aim of this project is to investigate whether APPL2 antisense oligonucleotide (ASO) can alleviate dietary-induced T2D and hypercholesterolemia. High-fat diet (HFD) was used to induce obesity and insulin resistance in mice. GalNAc-conjugated APPL2 ASO (GalNAc-APPL2-ASO) was used to silence hepatic APPL2 expression in C57/BL6J mice selectively. Glucose, lipid, and energy metabolism were monitored. Immunoblotting and quantitative PCR analysis showed that GalNAc-APPL2-ASO treatment selectively reduced APPL2 expression in the liver instead of other tissues, like adipose tissues, kidneys, muscle, and heart. The glucose tolerance test and insulin sensitivity test revealed that GalNAc-APPL2-ASO improved glucose tolerance and insulin sensitivity progressively. Blood chemistry analysis revealed that the mice treated with GalNAc-APPL2-ASO had significantly lower circulating levels of total cholesterol and LDL cholesterol. However, there was no difference in circulating levels of high-density lipoprotein (HDL) cholesterol, triglyceride, and free fatty acid between the mice treated with GalNac-APPL2-ASO and GalNAc-Control-ASO. No obvious effect on food intake, body weight, and liver injury markers after GalNAc-APPL2-ASO treatment was found, supporting its tolerability and safety. We showed that selectively silencing hepatic APPL2 alleviated insulin resistance and hypercholesterolemia and improved energy metabolism in the dietary-induced obese mouse model, indicating APPL2 as a therapeutic target for metabolic diseases.

Keywords: APPL2, antisense oligonucleotide, hypercholesterolemia, type 2 diabetes

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1698 Induction Motor Analysis Using LabVIEW

Authors: E. Ramprasath, P. Manojkumar, P. Veena

Abstract:

Proposed paper dealt with the modelling and analysis of induction motor based on the mathematical expression using the graphical programming environment of Laboratory Virtual Instrument Engineering Workbench (LabVIEW). Induction motor modelling with the mathematical expression enables the motor to be simulated with the various required parameters. Owing to the invention of variable speed drives study about the induction motor characteristics became complex.In this simulation motor internal parameter such as stator resistance and reactance, rotor resistance and reactance, phase voltage, frequency and losses will be given as input. By varying the speed of motor corresponding parameters can be obtained they are input power, output power, efficiency, torque induced, slip and current.

Keywords: induction motor, LabVIEW software, modelling and analysi, electrical and mechanical characteristics of motor

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1697 Using Lysosomal Immunogenic Cell Death to Target Breast Cancer via Xanthine Oxidase/Micro-Antibody Fusion Protein

Authors: Iulianna Taritsa, Kuldeep Neote, Eric Fossel

Abstract:

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

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

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