Search results for: laboratory molecular diagnostics

Authors: Amr A. El Hanafy, Yasser M. Saad, Saleh A. Alkarim, Hussein A. Almehdar, Elrashdy M. Redwan

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Camels are substantial providers of transport, milk, sport, meat, shelter, fuel, security and capital in many countries, particularly Saudi Arabia. Identification of animal breeds has progressed rapidly during the last decade. Advanced molecular techniques are playing a significant role in breeding or strain protection laws. On the other hand, fingerprinting of some molecular markers related to some productive traits in farm animals represents most important studies to our knowledge, which aim to conserve these local genetic resources, and to the genetic improvement of such local breeds by selective programs depending on gene markers. Milk records were taken two days in each week from female camels of Majahem, Safara, Wathaha, and Hamara breeds, respectively from different private farms in northern Jeddah, Riyadh and Alwagh governorates and average weekly yields were calculated. DNA sequencing for CSN2 gene was used for evaluating the genetic variations and calculating the genetic distance values among four Saudi camel populations which are Hamra(R), Safra(Y), Wadha(W) and Majaheim(M). In addition, this marker was analyzed for reconstructing the Neighbor joining tree among evaluating camel breeds. In respect to milk yield during winter season, result indicated that average weekly milk yield of Safara camel breed (30.05 Kg/week) is significantly (p < 0.05) lower than the other 3 breeds which ranged from 39.68 for Hamara to 42.42 Kg/week for Majahem, while there are not significant differences between these three breeds. The Neighbor Joining analysis that re-constructed based on DNA variations showed that samples are clustered into two unique clades. The first clade includes Y (from Y4 to Y18) and M (from M1, to M9). On the other hand, the second cluster is including all R (from R1 to R6) and W (from W1 to W6). The genetic distance values were equal 0.0068 (between the groups M&Y and R&W) and equal 0 (within each group).

Keywords: milk yield, beta-casein precursor (CSN2), Saudi camel, molecular markers

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Authors: Kobid Panthi, Suttisak Soralump, Suriyon Prempramote

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In June 27, 2014 slope movement was observed in upstream side of Khlong Pa Bon Dam, Thailand. The slide did not have any major catastrophic impact on the dam structure but raised a very important question; why did the slide occur after 10 years of operation? Various site investigations (Bore Hole Test, SASW, Echo Sounding, and Geophysical Survey), laboratory analysis and numerical modelling using SIGMA/W and SLOPE/W were conducted to determine the cause of slope movement. It was observed that the dam had undergone the greatest differential drawdown in its operational history in the year 2014 and was termed as the major cause of movement. From the laboratory tests, it was found that the shear strength of clay had decreased with a period of time and was near its residual value. The cyclic movement of water, i.e., reservoir filling and emptying was coined out to be the major cause for the reduction of shear strength. The numerical analysis was carried out using a modified cam clay (MCC) model to determine the strain softening behavior of the clay. The strain accumulation was observed in the slope with each reservoir cycle triggering the slope failure in 2014. It can be inferred that if there was no major drawdown in 2014, the slope would not have failed but eventually would have failed after a long period of time. If there was no major drawdown in 2014, the slope would not have failed. However, even if there hadn’t been a drawdown, it would have failed eventually in the long run.

Keywords: slope movement, strain softening, residual strength, modified cam clay

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Authors: Anju Elizbath Peter, S. M. Shiva Nagendra, Indumathi M.Nambi

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The deliberate fires initiated by dump managers and human scavengers to reduce the volume of waste and recovery of valuable metals/materials are common at municipal solid waste (MSW) disposal sites in developed country. A large amount of toxic gases released due to this act is responsible for the deterioration of regional and local air quality, which causes visibility impairment and acute respiratory diseases. The present study was aimed at the characterization of MSW and emission characteristics of burning of MSW in the laboratory. MSW samples were collected directly from the one of the open dumpsite located in Chennai city. Solid waste sampling and laboratory analysis were carried out according to American Society of Testing and Materials (ASTM) standards. Results indicated the values of moisture content, volatile solids (VS) and calorific values of solid waste samples were 16.67%,8%,9.17MJ/kg, respectively. The elemental composition showed that the municipal solid waste contains 25.84% of carbon, 3.69% of hydrogen, 1.57% of nitrogen and 0.26% of sulphur. The calorific value of MSW was found to be 9.17 MJ/Kg which is sufficient to facilitate self-combustion of waste. The characterization of emissions from the burning of 1 kg of MSW in the test chamber showed a total of 90 mg/kg of PM10 and 243 mg/kg of PM2.5. The current research study results will be useful for municipal authorities to formulate guideline and policy structure regarding the MSW management to reduce the impact of air emissions at an open dump site.

Keywords: characterization, MSW, open burning, PM10, PM2.5

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Authors: Hiroyuki Aoki

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The physical properties of polymer materials are dependent on the conformation and molecular motion of a polymer chain. Therefore, the structure and dynamic behavior of the single polymer chain have been the most important concerns in the field of polymer physics. However, it has been impossible to directly observe the conformation of the single polymer chain in a bulk medium. In the current work, the novel techniques to study the conformation and dynamics of a single polymer chain are proposed. Since a fluorescence method is extremely sensitive, the fluorescence microscopy enables the direct detection of a single molecule. However, the structure of the polymer chain as large as 100 nm cannot be resolved by conventional fluorescence methods because of the diffraction limit of light. In order to observe the single chains, we developed the labeling method of polymer materials with a photo-switchable dye and the super-resolution microscopy. The real-space conformational analysis of single polymer chains with the spatial resolution of 15-20 nm was achieved. The super-resolution microscopy enables us to obtain the three-dimensional coordinates; therefore, we succeeded the conformational analysis in three dimensions. The direct observation by the nanometric optical microscopy would reveal the detailed information on the molecular processes in the various polymer systems.

Keywords: polymer materials, single molecule, super-resolution techniques, conformation

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Authors: Anna Pajdak, Mateusz Kudasik, Norbert Skoczylas, Leticia Teixeira Palla Braga

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A substantial majority of gas transport and sorption researches into coal are carried out on samples that are free of stress. In natural conditions, coal occurs at considerable depths, which often exceed 1000 meters. In such conditions, coal is subjected to geostatic pressure. Thus, in natural conditions, the sorption capacity of coal subjected to geostatic pressure can differ considerably from the sorption capacity of coal, determined in laboratory conditions, which is free of stress. The work presents the results of filtration and sorption tests of gases in coal under confining pressure conditions. The tests were carried out on the author's device, which ensures: confining pressure regulation in the range of 0-30 MPa, isobaric gas pressure conditions, and registration of changes in sample volume during its gas saturation. Based on the conducted research it was found, among others, that the sorption capacity of coal relative to CO₂ was reduced by about 15% as a result of the change in the confining pressure from 1.5 MPa to 30 MPa exerted on the sample. The same change in sample load caused a significant, more than tenfold reduction in carbon permeability to CO₂. The results confirmed that a load of coal corresponding to a hydrostatic pressure of 1000 meters underground reduces its permeability and sorption properties. These results are so important that the effect of load on the sorption properties of coal should be taken into account in laboratory studies on the applicability of CO₂ Enhanced Coal Bed Methane Recovery (CO₂-ECBM) technology.

Keywords: coal, confining pressure, gas transport, sorption

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Authors: Natalie M. Wride, Xueyu Geng

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Subgrade soils of railway infrastructure are subjected to a significant number of load applications over their design life. The use of slab track on existing and future proposed rail links requires a reduced maintenance and repair regime for the embankment subgrade, due to restricted access to the subgrade soils for remediation caused by cyclic deformation. It is, therefore, important to study the deformation behaviour of soft cohesive subsoils induced as a result of long term cyclic loading. In this study, a series of oedometer tests and cyclic triaxial tests (10,000 cycles) have been undertaken to investigate the undrained deformation behaviour of soft kaolin. X-ray Computer Tomography (CT) scanning of the samples has been performed to determine the change in porosity and soil structure density from the sample microstructure as a result of the laboratory testing regime undertaken. Combined with the examination of excess pore pressures and strains obtained from the cyclic triaxial tests, the results are compared with an existing analytical solution for long term settlement considering repeated low amplitude loading. Modifications to the analytical solution are presented based on the laboratory analysis that shows good agreement with further test data.

Keywords: creep, cyclic loading, deformation, long term settlement, train loading

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Authors: Azene Tesfaye, Yohannes Petros, Habtamu Zeleke

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To evaluate genetic variation among Ethiopian lentil, laboratory experiment were conducted to screen 12 accessions of lentil (Lens culinaris M.) for salt tolerance. Seeds of 12 Lentil accessions were grown at laboratory (Petri dish) condition with different levels of salinity (0, 2, 4, and 8 dSm-1 NaCl) for 4 weeks. The experimental design was completely randomized design (CRD) in factorial combination with three replications. Data analysis was carried out using SAS software. Average germination time, germination percentage, seedling shoot and root traits, seedling shoot and root weight were evaluated. The two way ANOVA for varieties revealed statistically significant variation among lentil accession, NaCl level and their interactions (p<0.001) with respect to the entire parameters. It was found that salt stress significantly delays germination rate and decreases germination percentage, shoot and root length, seedling shoot and root weight of lentil accessions. The degree of decrement varied with accessions and salinity levels. Accessions 36120, 9235 and 36004 were better salt tolerant than the other accessions. As the result, it is recommended to be used as a genetic resource for the development of lentil accession and other very salt sensitive crop with improved germination under salt stress condition.

Keywords: accession, germination, lentil, NaCl, screening, seedling stage

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Authors: Apisaraporn Baicharoen, Prapasiri Pongprayoon

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Betaine aldehyde dehydrogenase 2 (BADH2) is an enzyme that inhibits the accumulation of 2-acetyl-1-pyrroline (2AP), a potent flavor compound in rice fragrance. BADH2 contains three domains (NAD-binding, substrate-binding, and oligomerization domains). It catalyzes the oxidation of amino aldehydes. The lack of BADH2 results in the formation of 2AP and consequently an increase in rice fragrance. To date, inadequate data on BADH2 structure and function are available. An insight into the nature of BADH2 can serve as one of key starting points for the production of high quality fragrant rice. In this study, we therefore constructed the homology model of BADH2 and employed 500-ns Molecular Dynamics simulations (MD) to primarily understand the structural and dynamic properties of BADH2. Initially, Ramachandran plot confirms the good quality of modeled protein structure. Principle Component Analysis (PCA) was also calculated to capture the protein dynamics. Among 3 domains, the results show that NAD binding site is found to be more flexible. Moreover, interactions from key amino acids (N162, E260, C294, and Y419) that are crucial for function are investigated.

Keywords: betaine aldehyde dehydrogenase 2, fragrant rice, homology modeling, molecular dynamics simulations

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Authors: Nicola G. G. Cecca

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In Italian IPSSEOAs, high schools that give a vocational education to students that will work in the field of Enogastronomy and Hotel Management, the course of Food Science allows the students to start and see food as a mixture of substances that they will transform during their profession. These substances are characterized not only by a chemical composition but also by a molecular structure that makes them nutritionally active. But the increasing number of new products proposed by Food Industry, the modern techniques of production and transformation, the innovative preparations required by customers have made many information reported in the most wide spread Food Science textbooks not up-to-date or too poor for the people who will work in catering sector. Often Authors offer information aged to Bohr’s Atomic Model and to the ‘Octet Rule’ proposed by G.N. Lewis to describe the Chemical Bond, without giving any reference to new as Orbital Atomic Model and Molecular Orbital Theory that, in the meantime, start to be old themselves. Furthermore, this antiquated information precludes an easy understanding of a wide range of properties of nutritive substances and many reactions in which the food constituents are involved. In this paper, our attention is pointed out to use GEOMAG™ to represent the dynamics with which the chemical bond is formed during the synthesis of the molecules. GEOMAG™ is a toy, produced by the Swiss Company Geomagword S.A., pointed to stimulate in children, aged between 6-10 years, their fantasy and their handling ability and constituted by metallic spheres and metallic magnetic bars coated by coloured plastic materials. The simulation carried out with GEOMAG™ is based on the similitude existing between the Coulomb’s force and the magnetic attraction’s force and in particular between the formulae with which they are calculated. The electrostatic force (F in Newton) that allows the formation of the chemical bond can be calculated by mean Fc = kc q1 q2/d2 where: q1 e q2 are the charge of particles [in Coulomb], d is the distance between the particles [in meters] and kc is the Coulomb’s constant. It is surprising to observe that the attraction’s force (Fm) acting between the magnetic extremities of GEOMAG™ used to simulate the chemical bond can be calculated in the same way by using the formula Fm = km m1 m2/d2 where: m1 e m2 represent the strength of the poles [A•m], d is the distance between the particles [m], km = μ/4π in which μ is the magnetic permeability of medium [N•A-2]. The magnetic attraction can be tested by students by trying to keep the magnetic elements of GEOMAG™ separate by hands or trying to measure by mean an appropriate dynamometric system. Furthermore, by using a dynamometric system to measure the magnetic attraction between the GEOMAG™ elements is possible draw a graphic F=f(d) to verify that the curve obtained during the simulation is very similar to that one hypnotized, around the 1920’s by Linus Pauling to describe the formation of H2+ in according with Molecular Orbital Theory.

Keywords: chemical bond, molecular orbital theory, magnetic attraction force, GEOMAG™

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Authors: Rumana Keyani, Haleema Sadia, Asia Nosheen, Rabia Naz, Humaira Yasmin, Sidra Zahoor

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Tomato is a significant crop of the world and is one of the staple foods of Pakistan. A vast number of plant pathogens from simple viruses to complex parasites cause diseases in tomatoes but fungal infection in our country is quite high. Sometimes the symptoms are too harsh destroying the crop altogether. Countries like our own with continuously increasing massive population and limited resources cannot afford such an economic loss. There is an array of morphological, genetic, biochemical and molecular processes involved in plant resistance mechanisms to biotic stress. The study of different metabolic pathways like Jasmonic acid (JA) pathways and most importantly signaling molecules like ROS/RNS and their redoxin enzymes i.e. TRX and NRX is crucial to disease management, contributing to healthy plant growth. So, improving tolerance in crop plants against biotic stresses is a dire need of our country and world as whole. In the current study, fungal pathogenic strains Alternaria solani and Rhizoctonia solani were used to inoculate tomatoes to check the defense responses of tomato plant against these pathogens at molecular as well as phenotypic level with jasmonic acid and spermidine pretreatment. All the growth parameters (root and shoot length, dry and weight root, shoot weight measured 7 days post-inoculation, exhibited that infection drastically declined the growth of the plant whereas jasmonic acid and spermidine assisted the plants to cope up with the infection. Thus, JA and Spermidine treatments maintained comparatively better growth factors. Antioxidant assays and expression analysis through real time quantitative PCR following time course experiment at 24, 48 and 72 hours intervals also exhibited that activation of JA defense genes and a polyamine Spermidine helps in mediating tomato responses against fungal infection when used alone but the two treatments combined mask the effect of each other.

Keywords: fungal infection, jasmonic acid defence, tomato, spermidine

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Authors: Mohammad Nazri Abdul Bahari, Nurshafika Mohd Sakeh, Siti Nor Akmar Abdullah

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Basal stem rot (BSR) is the most devastating disease in oil palm. Among the oil palm pathogenic fungi, the most prevalent and virulent species associated with BSR is Ganoderma boninense. Early detection of G. boninense attack in oil palm wherein physical symptoms has not yet appeared can offer opportunities to prevent the spread of the necrotrophic fungus. However, poor understanding of molecular defense responses and roles of antifungal metabolites in oil palm against G. boninense has complicated the resolving measures. Hence, characterization of defense-related molecular responses and production of antifungal compounds during early interaction with G. boninense is of utmost important. Four month-old oil palm (Elaeis guineensis) seedlings were artificially infected with G. boninense-inoculated rubber wood block via sitting technique. RNA of samples were extracted from roots and leaves tissues at 0, 3, 7 and 11 days post inoculation (d.p.i) followed with sequencing using RNA-Seq method. Differentially-expressed genes (DEGs) of oil palm-G. boninense interaction were identified, while changes in metabolite profile will be scrutinized related to the DEGs. The RNA-Seq data generated a total of 113,829,376 and 313,293,229 paired-end clean reads from untreated (0 d.p.i) and treated (3, 7, 11 d.p.i) samples respectively, each with two biological replicates. The paired-end reads were mapped to Elaeis guineensis reference genome to screen out non-oil palm genes and subsequently generated 74,794 coding sequences. DEG analysis of phytohormone biosynthetic genes in oil palm roots revealed that at p-value ≤ 0.01, ethylene and jasmonic acid may act in antagonistic manner with salicylic acid to coordinate defense response at early interaction with G. boninense. Findings on metabolite profiling of G. boninense-infected oil palm roots and leaves are hoped to explain the defense-related compounds elicited by Elaeis guineensis in response to G. boninense colonization. The study aims to shed light on molecular defense response of oil palm at early interaction with G. boninense and promote prevention measures against Ganoderma infection.

Keywords: Ganoderma boninense, metabolites, phytohormones, RNA-Seq

Procedia PDF Downloads 254

Authors: Elmira Ghanbari, Jan Van Esch, Stephen J. Picken, Sahil Aggarwal

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Organogels are formed by the assembly of low molecular weight gelators (LMWG) into fibrous structures. The assembly of these molecules into crystalline fibrous structures occurs as a result of reversible interactions such as π-stacking, hydrogen-bonding, and van der Waals interactions. Bisamide organogelators with two amide groups have been used as one of LMWGs which show efficient assembly behavior via hydrogen bonding for network formation, the formation of a crystalline network for solvent entrapment. In this study, different bisamide gelators with different lengths of alkyl chains have been added to the bisamide parent gels. The effect of the addition of bisamide additives on the gelation of bisamide gels is described. Investigation of the thermal properties of the gels by differential scanning calorimetry and dropping ball techniques indicated that the bisamide gels can be formed by the addition of a high concentration of the second bisamide components. The microstructure of the gels with different gelator components has been visualized with scanning electron microscopy (SEM) which has shown systematic woven, platelet-like, and a combination of those morphologies for different gels. Examining the addition of a range of bisamide additives with different structural characteristics than the parent bisamide gels has confirmed the effect of the molecular structure on the morphology of the bisamide gels and their final properties.

Keywords: bisamide organogelator additives, gel morphology, gel properties, self-assembly

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Authors: Abdulwahab Kammon, Tan Sheau Wei, Abdul Rahman Omar, Abdunaser Dayhum, Ibrahim Eldghayes, Monier Sharif

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Infectious bronchitis virus (IBV) is a very dynamic and evolving virus which causing major economic losses to the global poultry industry. Recently, the Libyan poultry industry faced severe outbreak of respiratory distress associated with high mortality and dramatic drop in egg production. Tracheal and cloacal swabs were analyzed for several poultry viruses. IBV was detected using SYBR Green I real-time PCR detection based on the nucleocapsid (N) gene. Sequence analysis of the partial N gene indicated high similarity (~ 94%) to IBV strain 3382/06 that was isolated from Taiwan. Even though the IBV strain 3382/06 is more similar to that of the Mass type H120, the isolate has been implicated associated with intertypic recombinant of 3 putative parental IBV strains namely H120, Taiwan strain 1171/92 and China strain CK/CH/LDL/97I. Complete sequencing and antigenicity studies of the Libya IBV strains are currently underway to determine the evolution of the virus and its importance in vaccine induced immunity. In this paper, we documented for the first time the presence of possibly variant IBV strain from Libya which required a dramatic change in the vaccination program.

Keywords: Libya, infectious bronchitis, molecular characterization, viruses, vaccine

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Authors: Ana Flavia Belchior De Andrade

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Forensic laboratories all over the world face a great challenge to overcame waiting time and backlog in many different areas. Many aspects contribute to this situation, such as an increase in drug complexity, increment in the number of exams requested and cuts in funding limiting laboratories hiring capacity. Altogether, those facts pose an essential challenge for forensic chemistry laboratories to keep both quality and time of response within an acceptable period. In this paper we will analyze how the backlog affects test results and, in the end, the whole judicial system. In this study data from marijuana samples seized by the Federal District Civil Police in Brazil between the years 2013 and 2017 were tabulated and the results analyzed and discussed. In the last five years, the number of petitioned exams increased from 822 in February 2013 to 1358 in March 2018, representing an increase of 32% in 5 years, a rise of more than 6% per year. Meanwhile, our data shows that the number of performed exams did not grow at the same rate. Product numbers are stationed as using the actual technology scenario and analyses routine the laboratory is running in full capacity. Marijuana detection is the most prevalence exam required, representing almost 70% of all exams. In this study, data from 7,110 (seven thousand one hundred and ten) marijuana samples were analyzed. Regarding waiting time, most of the exams were performed not later than 60 days after receipt (77%). Although some samples waited up to 30 months before being examined (0,65%). When marijuana´s exam is delayed we notice the enlargement of inconclusive results using thin-layer chromatography (TLC). Our data shows that if a marijuana sample is stored for more than 18 months, inconclusive results rise from 2% to 7% and when if storage exceeds 30 months, inconclusive rates increase to 13%. This is probably because Cannabis plants and preparations undergo oxidation under storage resulting in a decrease in the content of Δ9-tetrahydrocannabinol ( Δ9-THC). An inconclusive result triggers other procedures that require at least two more working hours of our analysts (e.g., GC/MS analysis) and the report would be delayed at least one day. Those new procedures increase considerably the running cost of a forensic drug laboratory especially when the backlog is significant as inconclusive results tend to increase with waiting time. Financial aspects are not the only ones to be observed regarding backlog cases; there are also social issues as legal procedures can be delayed and prosecution of serious crimes can be unsuccessful. Delays may slow investigations and endanger public safety by giving criminals more time on the street to re-offend. This situation also implies a considerable cost to society as at some point, if the exam takes a long time to be performed, an inconclusive can turn into a negative result and a criminal can be absolved by flawed expert evidence.

Keywords: backlog, forensic laboratory, quality management, accreditation

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Authors: Hamza Javar Magnier, Leslie V. Woodcock

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It has been reported that at temperatures above the critical there is no “continuity of liquid and gas”, as originally hypothesized by van der Waals. Rather, both gas and liquid phases, with characteristic properties as such, extend to supercritical temperatures. Each phase is bounded by the locus of a percolation transition, i.e. a higher-order thermodynamic phase change associated with percolation of gas clusters in a large void, or liquid interstitial vacancies in a large cluster. Between these two-phase bounds, it is reported there exists a mesophase that resembles an otherwise homogeneous dispersion of gas micro-bubbles in liquid (foam) and a dispersion of liquid micro-droplets in gas (mist). Such a colloidal-like state of a pure one-component fluid represents a hitherto unchartered equilibrium state of matter besides pure solid, liquid or gas. Here we provide compelling evidence, from molecular dynamics (MD) simulations, for the existence of this supercritical mesophase and its colloidal nature. We report preliminary results of computer simulations for a model fluid using a simplistic representation of atoms or molecules, i.e. a hard-core repulsion with an attraction so short that the atoms are referred to as “adhesive spheres”. Molecular clusters, and hence percolation transitions, are unambiguously defined. Graphics of color-coded clusters show colloidal characteristics of the supercritical mesophase.

Keywords: critical phenomena, mesophase, supercritical, square-well, critical parameters

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Authors: B. Gonzalez-Yebra, H. Barajas, P. Palomares, M. Hernandez, O. Torres, M. Ayala, A. L. González, G. Vazquez-Ortiz, M. L. Guzman

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Leukemia arises by molecular alterations of the normal hematopoietic stem cell (HSC) transforming it into a leukemic stem cell (LSC) with high cell proliferation, self-renewal, and cell differentiation. Chronic myeloid leukemia (CML) originates from an LSC-leading to elevated proliferation of myeloid cells and acute lymphoblastic leukemia (ALL) originates from an LSC development leading to elevated proliferation of lymphoid cells. In both cases, LSC can be identified by multicolor flow cytometry using several antibodies. However, to date, LSC levels in peripheral blood (PB) are not established well enough in ALL and CML patients. On the other hand, the detection of the minimal residue disease (MRD) in leukemia is mainly based on the identification of the mRNA bcr-abl gene in CML patients and some other genes in ALL patients. There is no a properly biomarker to detect MDR in both types of leukemia. The objective of this study was to determine mRNA bcr-abl and the percentage of LSC in peripheral blood of patients with CML and ALL and identify a possible association between the amount of LSC in PB and clinical data. We included in this study 19 patients with Leukemia. A PB sample was collected per patient and leukocytes were obtained by Ficoll gradient. The immunophenotype for LSC CD34+ was done by flow cytometry analysis with CD33, CD2, CD14, CD16, CD64, HLA-DR, CD13, CD15, CD19, CD10, CD20, CD34, CD38, CD71, CD90, CD117, CD123 monoclonal antibodies. In addition, to identify the presence of the mRNA bcr-abl by RT-PCR, the RNA was isolated using TRIZOL reagent. Molecular (presence of mRNA bcr-abl and LSC CD34+) and clinical results were analyzed with descriptive statistics and a multiple regression analysis was performed to determine statistically significant association. In total, 19 patients (8 patients with ALL and 11 patients with CML) were analyzed, 9 patients with de novo leukemia (ALL = 6 and CML = 3) and 10 under treatment (ALL = 5 and CML = 5). The overall frequency of mRNA bcr-abl was 31% (6/19), and it was negative in ALL patients and positive in 80% in CML patients. On the other hand, LSC was determined in 16/19 leukemia patients (%LSC= 0.02-17.3). The Novo patients had higher percentage of LSC (0.26 to 17.3%) than patients under treatment (0 to 5.93%). The amount of LSC was significantly associated with the amount of LSC were: absence of treatment, the absence of splenomegaly, and a lower number of leukocytes, negative association for the clinical variables age, sex, blasts, and mRNA bcr-abl. In conclusion, patients with de novo leukemia had a higher percentage of circulating LSC than patients under treatment, and it was associated with clinical parameters as lack of treatment, absence of splenomegaly and a lower number of leukocytes. The mRNA bcr-abl detection was only possible in the series of patients with CML, and molecular detection of LSC could be identified in the peripheral blood of all leukemia patients, we believe the identification of circulating LSC may be used as biomarker for the detection of the MRD in leukemia patients.

Keywords: stem cells, leukemia, biomarkers, flow cytometry

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Authors: D. Garg, S. Luthra, A. Haleem

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Due to growing concern about environmental and social consequences throughout the world, a need has been felt to incorporate sustainability concepts in conventional manufacturing. This paper is an attempt to identify and evaluate drivers in implementing sustainable manufacturing in Indian context. Nine possible drivers for successful implementation of sustainable manufacturing have been identified from extensive review. Further, Decision Making Trial and Evaluation Laboratory (DEMATEL) approach has been utilized to evaluate and categorize these identified drivers for implementing sustainable manufacturing in to the cause and effect groups. Five drivers (Societal Pressure and Public Concerns; Regulations and Government Policies; Top Management Involvement, Commitment and Support; Effective Strategies and Activities towards Socially Responsible Manufacturing and Market Trends) have been categorized into the cause group and four drivers (Holistic View in Manufacturing Systems; Supplier Participation; Building Sustainable culture in Organization; and Corporate Image and Benefits) have been categorized into the effect group. “Societal Pressure and Public Concerns” has been found the most critical driver and “Corporate Image and Benefits” as least critical or the most easily influenced driver to implementing sustainable manufacturing in Indian context. This paper may surely help practitioners in better understanding of these drivers and their priorities towards effective implementation of sustainable manufacturing.

Keywords: drivers, decision making trial and evaluation laboratory (DEMATEL), India, sustainable manufacturing

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Authors: Seyed Hassan Moosa-kazemi, Jalal Mohammadi Soleimani, Hassan Vatandoost, Mohammad Hassan Shirazi, Sara Hajikhani, Roonak Bakhtiari, Morteza Akbari, Siamak Hydarzadeh

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Malaria is an infectious disease and considered most important health problems in the southeast of Iran. Iran is elimination malaria phase and new tool need to vector control. Paraterasgenesis is a new way to cut of life cycle of the malaria parasite. In this study, the microflora of the surface and gut of various stages of Anopheles fluviatilis James as one of the important malaria vector was studied using biochemical and molecular techniques during 2013-2014. Twelve bacteria species were found including; Providencia rettgeri, Morganella morganii, Enterobacter aerogenes, Pseudomonas oryzihabitans, Citrobacter braakii، Citrobacter freundii، Aeromonas hydrophila، Klebsiella oxytoca, Citrobacter koseri, Serratia fonticola، Enterobacter sakazakii and Yersinia pseudotuberculosis. The species of Alcaligenes faecalis, Providencia vermicola and Enterobacter hormaechei were identified in various stages of the vector and confirmed by biochemical and molecular techniques. We found Providencia rettgeri proper candidate for paratransgenesis.

Keywords: Anopheles fluviatilis, bacteria, malaria, Paraterasgenesis, Southern Iran

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Authors: Gehan Aouf, Diala Tabbal, Abd El Rahim Sabsabi, Rashad Aouf

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The aim of this study is to assess the viability of utilizing Municipal Solid Waste Incineration Fly Ash (MSWIFA) with Ordinary Portland cement as soil reinforcement materials for geotechnical engineering applications. A detailed experimental program is carried out, followed by analysis of results. Soil samples were prepared by adding Cement to MSWIFA-soil mix at different percentages. Then, a series of laboratory tests were performed, namely: Sieve analysis, Atterberg limits tests, Unconfined compression test, and Proctor tests. A parametric study is conducted to investigate the effect of adding the cement at different percentages on the unconfined compression strength, maximum dry density, and optimum moisture content of clayey soil-MSWIFA The variation of contents of admixtures were 10%, 20%, and 30% for MSWIFA by dry total weight of soil and 10%, 15%, and 20% for Portland cement by dry total weight of the mix. The test results reveal that adding MSWIFA to the soil up to 20% increased the MDD of the mixture and decreased the OMC, then an opposite trend for results were found when the percentage of MSWIFA exceeds 20%. This is due to the low specific gravity of MSWIFA and to the greater water absorption of MSWIFA. The laboratory tests also indicate that the UCS values were found to be increased for all the mixtures with curing periods of 7, 14, and 28 days. It is also observed that the cement increased the strength of the finished product of the mix of soil and MSWIFA.

Keywords: clayey soil, cement, MSWIFA, unconfined compression strength

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Authors: Thangaraj Arasakumar, Athar Ata, Palathurai Subramaniam Mohan

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A series of novel 4-quinolone-3-carboxylic acid grafted spiropyrrolidines as new type of antibacterial agents were synthesized via multicomponent 1,3-dipolar cycloaddition reaction of an azomethine ylides with a newly prepared (E)-4-oxo-6-(3-phenyl-acryloyl)-1,4-dihydro-quinoline-3-carboxylic acids in high regioselectivity with good yields. The structure of cycloadduct characterized by FT IR, mass, 1H, 13C, 2D NMR techniques and elemental analysis. Structure and spectrometry of compound 8a has been investigated theoretically by using HF and DFT approach at B3LYP, M05-2x/6-31G* levels of theories. The optimized geometries and calculated vibrational frequencies are evaluated via comparison with experimental values. A good agreement is found between the measured and calculated values. The DFT studies support the molecular mechanism of this cycloaddition reaction and determine the molecular electrostatic potential and thermodynamic properties. Furthermore, the antibacterial activities of synthesized compounds were evaluated against Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis) and Gram-negative bacteria strains (Escherichia coli, Klebsiella pneumoniae). Among 21 compounds screened, 8f and 8p were found to be more active against tested bacteria.

Keywords: antibacterial activity, azomethine ylide, DFT calculation, spirooxindole

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Authors: S. Taghavi Kalajahi, A. Koerdt, T. Lund Skovhus

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Cathodic protection (CP) is an electrochemical method to control and manage corrosion in different industries and environments. CP which is widely used, especially in buried and sub-merged environments, which both environments are susceptible to microbiologically influenced corrosion (MIC). Most of the standards recommend performing CP using -800 mV, however, if MIC threats are high or sulfate reducing bacteria (SRB) is present, the recommendation is to use more negative potentials for adequate protection of the metal. Due to the lack of knowledge and research on the effectiveness of CP on MIC, to the author’s best knowledge, there is no information about what MIC threat is and how much more negative potentials should be used enabling adequate protection and not overprotection (due to hydrogen embrittlement risk). Recently, the development and cheaper price of molecular microbial methods (MMMs) open the door for more effective investigations on the corrosion in the presence of microorganisms, along with other electrochemical methods and surface analysis. In this work, using MMMs, the gene expression of SRB biofilm under different potentials of CP will be investigated. The specific genes, such as pH buffering, metal oxidizing, etc., will be compared at different potentials, enabling to determine the precise potential that protect the metal effectively from SRB. This work is the initial step to be able to standardize the recommended potential under MIC condition, resulting better protection for the infrastructures.

Keywords: cathodic protection, microbiologically influenced corrosion, molecular microbial methods, sulfate reducing bacteria

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Authors: El-Sohaimy Sobhy A., Androsova Natalia, Toshev Abuvali Djabarovec

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The conditions of the isolation of proteins from the hemp seeds were optimized in the current work. Moreover, the physicochemical and functional properties of hemp protein isolate were evaluated for its potential application in food manufacturing. The elastin protein is the most predominant protein in the protein profile with a molecular weight of 58.1 KDa, besides albumin, with a molecular weight of 31.5 KDa. The FTIR spectrum detected the absorption peaks of the amide I in 1750 and 1600 cm⁻¹, which pointed to C=O stretching while N-H was stretching at 1650-1580 cm⁻¹. The peak at 3250 was related to N-H stretching of primary aliphatic amine (3400-3300 cm⁻¹), and the N-H stretching for secondary (II) amine appeared at 3350-3310 cm⁻¹. Hemp protein isolate (HPI) was showed high content of arginine (15.52 g/100 g), phenylalanine+tyrosine (9.63 g/100 g), methionine + cysteine (5.49 g/100 g), leucine + isoleucine (5.21 g/100 g) and valine (4.53 g/100 g). It contains a moderate level of threonine (3.29 g/100 g) and lysine (2.50 g/100 g), with the limiting amino acid being a tryptophan (0.22 g/100 g HPI). HPI showed high water-holding capacity (4.5 ± 2.95 ml/g protein) and oil holding capacity (2.33 ± 1.88 ml/g) values. The foaming capacity of HPI was increased with increasing the pH values to reach the maximum value at pH 11 (67.23±3.20 %). The highest emulsion ability index of HPI was noted at pH 9 (91.3±2.57 m2/g) with low stability (19.15±2.03).

Keywords: Cannabis sativa ssp., protein isolate, isolation conditions, amino acid composition, chemical properties, functional properties

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Authors: Menglim Hoy, Suksun Horpibulsuk, Arul Arulrajah

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The results of laboratory investigation of recycled asphalt pavement (RAP) – fly ash (FA) based geopolymer as a base material is presented in this paper. An alkaline activator, the mixture of NaOH and Na₂SiO₃, is used to synthesis RAP-FA based geopolymer. RAP-FA with water (RAP-FA blend) prepared as a control material. The strength develops and the strength against wet-dry was determined by the unconfined compression strength (UCS) test, then the microstructural properties were examined by scanning electron microscopy (SEM) and X-ray Diffraction (XRD) analysis. The toxicity characteristic leaching procedure (TCLP) test is conducted to measure its leachability of heavy metal. The results show both the RAP-FA blend and geopolymer can be used as a base course as its UCS values meet the minimum strength requirement specified by the Department of Highway, Thailand. The durability test results show the UCS of these materials increases with increasing the number of wet-dry cycles, reaching its peak at six wet-dry cycles. The XRD and SEM analyses indicate strength development of the RAP-FA blend occurs due to chemical reaction between a high Calcium in RAP with a high Silica and Alumina in FA led to producing calcium aluminate hydrate formation. The strength development of the RAP-FA geopolymer occurred resulted from the polymerization reaction. The TCLP results demonstrate there is no environmental risk of these stabilized materials. Furthermore, FA based geopolymer can reduce the leachability of heavy metal in the RAP-FA blend.

Keywords: recycled asphalt pavement, geopolymer, heavy metal, microstructure

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Authors: Michele Gattullo, Vito M. Manghisi, Alessandro Evangelista, Enricoandrea Laviola

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In this work, we explored the use of Augmented Reality (AR) to support students in laboratory lectures in Distance Learning (DL), designing an application that proved to be ready for use next semester. AR could help students in the understanding of complex concepts as well as increase their motivation in the learning process. However, despite many prototypes in the literature, it is still less used in schools and universities. This is mainly due to the perceived limited advantages to the investment costs, especially regarding changes needed in the teaching modalities. However, with the spread of epidemiological emergency due to SARS-CoV-2, schools and universities were forced to a very rapid redefinition of consolidated processes towards forms of Distance Learning. Despite its many advantages, it suffers from the impossibility to carry out practical activities that are of crucial importance in STEM ("Science, Technology, Engineering e Math") didactics. In this context, AR perceived advantages increased a lot since teachers are more prepared for new teaching modalities, exploiting AR that allows students to carry on practical activities on their own instead of being physically present in laboratories. In this work, we designed an AR application for the support of engineering students in the understanding of assembly drawings of complex machines. Traditionally, this skill is acquired in the first years of the bachelor's degree in industrial engineering, through laboratory activities where the teacher shows the corresponding components (e.g., bearings, screws, shafts) in a real machine and their representation in the assembly drawing. This research aims to explore the effectiveness of AR to allow students to acquire this skill on their own without physically being in the laboratory. In a preliminary phase, we interviewed students to understand the main issues in the learning of this subject. This survey revealed that students had difficulty identifying machine components in an assembly drawing, matching between the 2D representation of a component and its real shape, and understanding the functionality of a component within the machine. We developed a mobile application using Unity3D, aiming to solve the mentioned issues. We designed the application in collaboration with the course professors. Natural feature tracking was used to associate the 2D printed assembly drawing with the corresponding 3D virtual model. The application can be displayed on students’ tablets or smartphones. Users could interact with selecting a component from a part list on the device. Then, 3D representations of components appear on the printed drawing, coupled with 3D virtual labels for their location and identification. Users could also interact with watching a 3D animation to learn how components are assembled. Students evaluated the application through a questionnaire based on the System Usability Scale (SUS). The survey was provided to 15 students selected among those we participated in the preliminary interview. The mean SUS score was 83 (SD 12.9) over a maximum of 100, allowing teachers to use the AR application in their courses. Another important finding is that almost all the students revealed that this application would provide significant power for comprehension on their own.

Keywords: augmented reality, distance learning, STEM didactics, technology in education

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Authors: Karim Saeidi

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Safflower capsule fly, Acanthiophilus helianthi Rossi (Diptera: Tephritidae), is a key pest of safflower in Iran. The toxicity of Methidathion, Malathion, Deltamethrin, and Lufenuron to adult males and females of Acanthiophilus helianthi was studied under laboratory conditions. Malathion was the most toxic among the tested compounds followed by Methidathion, Lufenuron, and Deltamethrin to Acanthiophilus helianthi at 24 h post treatment, the respective LC50 values were 0.40 ppm, 0.68 ppm, 10.99 ppm, and 11.75 ppm for males and 0.46 ppm, 0.97 ppm, 13.45 ppm, and 16.32 ppm for females. At 48 h post treatment, Malathion was the most toxic followed by Methidathion, Deltamethrin, and Lufenuron to Acanthiophilus helianthi, LC50 values were 0.08 ppm, 0.54 ppm, 1.80 ppm, and 1.96 ppm for males and 0.34 ppm, 0.64 ppm, 1.88 ppm, and 2.37 ppm for females. At 72 h post treatment, Malathion was the most toxic followed by Methidathion, Lufenuron, and Deltamethrin to Acanthiophilus helianthi LC50 values were 0.04 ppm, 0.33 ppm, 0.44 ppm, and 0.71 ppm for males and 0.09 ppm, 0.36 ppm, 0.75 ppm, and 0.82 ppm for females. It is observed that LC50 values for treated adult females increased more than in the treated adult males at 24 h, 48 h, and 72 h post treatment. It means that the adult males were more susceptible to the tested insecticides than the adult females.

Keywords: safflower, Methidathion, Deltamethrin, Lufenuron, Malathion, Tephritidae, safflower capsule fly, Acanthiophilus helianthi

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Authors: Amir Halabianfar, Jamshid Razmjoo

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In order to evaluate the competitive effects of Lambsqua on the germination and early growth of two wheat (Triticum aestivum L.) varieties, an experiment was conducted in laboratory conditions in researches of agronomy, College of agriculture, Isfahan University of Technology in 2015. A laboratory experiment was conducted on a factorial arrangement in a randomized complete design with four replications. Testing factors include two wheat cultivars (Flat and Atila -4) and three level of Lambsqua (Chenopodium album) extract (30, 60 and 90 percent) plus control with no extract. Twenty-five seeds of each wheat varieties were placed in petri dish, then the root extract of lambsqua, which was prepared previously at three levels, was poured on the seeds in each petri dish. The result showed that allelopathic effect of Lambsquarter on germination, root, and shoot dry weight of two varieties was highly significant. Among varieties, the Atila–4 showed minimum germination at 60% while the Flat showed minimum germination at 90% concentration. In case of root dry weight, Atila–4 was more suppressed as compared to Flat at 60% concentration but at 90% concentration, the both wheat varieties were reduced non-significantly. Shoot dry weight of Flat were decreased non-significantly concentrations except Atila -4 that was more reduced at 60 % than 90% concentration.

Keywords: allelopathy, Chenopodium album, extract, germination, wheat, early growth

Procedia PDF Downloads 176

Authors: H. Merhni, A. Sbiti, I. Ratbi, A. Sefiani

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The proximal spinal muscular atrophy (SMA) is a group of neuromuscular disorders characterized by progressive muscle weakness due to the degeneration and loss of anterior motor neurons of the spinal cord. Depending on the age of onset of symptoms and their evolution, four types of SMA, varying in severity, result in a mutations of the SMN gene (survival of Motor neuron). We have analyzed the DNA of 295 patients referred to our genetic counseling; since January 1996 until October 2014; for suspected SMA. The homozygous deletion of exon 7 of the SMN gene was found in 133 patients; of which, 40.6% were born to consanguineous parents. In countries like Morocco, where the frequency of heterozygotes for SMA is high, genetic testing should be offered as first-line and, after careful clinical assessment, especially in newborns and infants with congenital hypotonia unexplained and prognosis compromise. The molecular diagnosis of SMA allows a quick and certainly diagnosis, provide adequate genetic counseling for families at risk and suggest, for couples who want prenatal diagnosis. The analysis of the SMN gene is a perfect example of genetic testing with an excellent cost/benefit ratio that can be of great interest in public health, especially in low-income countries. We emphasize in this work for the benefit of the generalization of molecular diagnosis of SMA by the technique of PCR-enzymatic digestion in other centers in Morocco.

Keywords: Exon7, PCR-digestion, SMA, SMN gene

Procedia PDF Downloads 237

Authors: Purva Mishra, Rajesh Potlia, Kuljeet Singh Sandhu

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The role of glycyl residues in the protein structure has lingered within the research community for the last several decades. Glycyl residue is the only amino acid that is achiral due to the lack of a side chain and can, therefore, exhibit Ramachandran conformations that are disallowed for L-amino acids. The structural and functional significance of glycyl residues with L-disallowed conformation, however, remains obscure. Through statistical analysis of various datasets, we found that the glycyls with L-disallowed conformations are over-represented at disease-associated sites and tend to be evolutionarily conserved. The mutations of L-disallowed glycyls tend to destabilize the native conformation, reduce protein solubility, and promote inter-molecular aggregations. We uncovered a structural motif referred to as “β-crescent” formed around the L-disallowed glycyl, which prevents β-sheet aggregation by disrupting the alternating pattern of β-pleats. The L-disallowed conformation of glycyls also holds predictive power to infer the pathogenic missense variants. Altogether, our observations highlight that the L-disallowed conformation of glycyls is selected to facilitate native folding and prevent inter-molecular aggregations. The findings may also have implications for designing more stable proteins and prioritizing the genetic lesions implicated in diseases.

Keywords: Ramachandran plot, β-sheet, protein stability, protein aggregation

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Authors: Fantu Lombamo Untiso, Sylvia Murphy, Emily Pieracci

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Background: Rabies is a highly fatal viral disease of all warm-blooded animals including human globally. However, effective rabies control program still remains to be a reality and needs to be strengthened. Objective: Reviewing of recorded data and analyzing it to generate information on the status of rabies in Addis Ababa in the year 2012/13. Methods: A retrospective data were used from the Ethiopian Public Health Institute rabies case record book registered in the year 2012/13. Results: Among 1357 suspected rabid animals clinically examined; only 8.84% were positive for rabies. Out of 216 animal brains investigated in the laboratory with Fluorescent Antibody Technique, 55.5% were confirmed rabies positive. Among the laboratory confirmed positive rabies cases, high percentage of the animals came from Yeka (20%) and lower number from Kirkos subcity (3.3%). Out of 1149 humans who came to the institute seeking anti-rabies post-exposure prophylaxis, 85.65% and 7.87% of them were exposed to suspected dogs and cats respectively. 3 human deaths due to rabies were reported in the year after exposure to dog bite of unknown vaccination status. Conclusion: The principal vector of rabies in Addis Ababa is dog. Effective rabies management and control based on confirmed cases and mass-immunization and control of stray dog populations is recommended.

Keywords: Addis Ababa, exposure, rabies, surveillance

Procedia PDF Downloads 172

Authors: Neşe Işık, Şenol Altıok, Galip Devrim Eryılmaz, Aydın durukan, Hasan Özgür Daş

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In this study, an active landslide zone affecting the road alignment on the Tortum-Uzundere (Erzurum/Turkey) highway was investigated. Due to the landslide movement, problems have occurred in the existing road pavement, which has caused both safety problems and reduced driving comfort in the operation of the road. In order to model the landslide, drilling, geophysical and inclinometer studies were carried out in the field within the scope of ground investigation. Laboratory tests were carried out on soil and rock samples obtained from the borings. When the drilling and geophysical studies were evaluated together, it was determined that the study area has a complex geological structure. In addition, according to the inclinometer results, the direction and speed of movement of the landslide mass were observed. In order to create an idealized geological profile, all field and laboratory studies were evaluated together and then the sliding surface of the landslide was determined by back analysis method. According to the findings obtained, it was determined that the landslide was massively large, and the movement occurred had a deep sliding surface. As a result of the numerical analyses, it was concluded that the Slope angle reduction is the most economical and environmentally friendly method for the control of the landslide mass.

Keywords: landslide, geotechnical methods, geophysics, monitoring, highway

Procedia PDF Downloads 62