Search results for: interspecific hybridization
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 159

Search results for: interspecific hybridization

69 Development of Filling Material in 3D Printer with the Aid of Computer Software for Supported with Natural Zeolite for the Removal of Nitrogen and Phosphorus

Authors: Luís Fernando Cusioli, Leticia Nishi, Lucas Bairros, Gabriel Xavier Jorge, Sandro Rogério Lautenschalager, Celso Varutu Nakamura, Rosângela Bergamasco

Abstract:

Focusing on the elimination of nitrogen and phosphorus from sewage, the study proposes to face the challenges of eutrophication and to optimize the effectiveness of sewage treatment through biofilms and filling produced by a 3D printer, seeking to identify the most effective Polylactic Acid (PLA), Acrylonitrile Butadiene Styrene (ABS). The study also proposes to evaluate the nitrification process in a Submerged Aerated Biological Filter (FBAS) on a pilot plant scale, quantifying the removal of nitrogen and phosphorus. The experiment will consist of two distinct phases, namely, a bench stage and the implementation of a pilot plant. During the bench stage, samples will be collected at five points to characterize the microbiota. Samples will be collected, and the microbiota will be investigated using Fluorescence In Situ Hybridization (FISH), deepening the understanding of the performance of biofilms in the face of multiple variables. In this context, the study contributes to the search for effective solutions to mitigate eutrophication and, thus, strengthen initiatives to improve effluent treatment.

Keywords: eutrophication, sewage treatment, biofilms, nitrogen and phosphorus removal, 3d printer, environmental efficiency

Procedia PDF Downloads 63
68 Ab Initio Study of Co2ZrGe and Co2NbB Full Heusler Compounds

Authors: A. Abada, S. Hiadsi, T. Ouahrani, B. Amrani, K. Amara

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Using the first-principles full-potential linearized augmented plane wave plus local orbital (FP-LAPW+lo) method based on density functional theory (DFT), we have investigated the electronic structure and magnetism of some Co2- based full Heusler alloys, namely Co2ZrGe and Co2NbB. The calculations show that these compounds are to be half-metallic ferromagnets (HMFs) with a total magnetic moment of 2.000 µB per formula unit, well consistent with the Slater-Pauling rule. Our calculations show indirect band gaps of 0.58 eV and 0.47 eV in the minority spin channel of density of states (DOS) for Co2ZrGe and Co2NbB, respectively. Analysis of the DOS and magnetic moments indicates that their magnetism is mainly related to the d-d hybridization between the Co and Zr (or Nb) atoms. The half metallicity is found to be robust against volume changes and the two alloys kept a 100% of spin polarization at the Fermi level. In addition, an atom inside molecule AIM formalism and an electron localization function ELF were also adopted to study the bonding properties of these compounds, building a bridge between their electronic and bonding behavior. As they have a good crystallographic compatibility with the lattice of semiconductors used industrially and negative calculated cohesive energies with considerable absolute values these two alloys could be promising magnetic materials in the spintronics field.

Keywords: half-metallic ferromagnets, full Heusler alloys, magnetic properties, electronic properties

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67 Sensitivity Enhancement in Graphene Based Surface Plasmon Resonance (SPR) Biosensor

Authors: Angad S. Kushwaha, Rajeev Kumar, Monika Srivastava, S. K. Srivastava

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A lot of research work is going on in the field of graphene based SPR biosensor. In the conventional SPR based biosensor, graphene is used as a biomolecular recognition element. Graphene adsorbs biomolecules due to carbon based ring structure through sp2 hybridization. The proposed SPR based biosensor configuration will open a new avenue for efficient biosensing by taking the advantage of Graphene and its fascinating nanofabrication properties. In the present study, we have studied an SPR biosensor based on graphene mediated by Zinc Oxide (ZnO) and Gold. In the proposed structure, prism (BK7) base is coated with Zinc Oxide followed by Gold and Graphene. Using the waveguide approach by transfer matrix method, the proposed structure has been investigated theoretically. We have analyzed the reflectance versus incidence angle curve using He-Ne laser of wavelength 632.8 nm. Angle, at which the reflectance is minimized, termed as SPR angle. The shift in SPR angle is responsible for biosensing. From the analysis of reflectivity curve, we have found that there is a shift in SPR angle as the biomolecules get attached on the graphene surface. This graphene layer also enhances the sensitivity of the SPR sensor as compare to the conventional sensor. The sensitivity also increases by increasing the no of graphene layer. So in our proposed biosensor we have found minimum possible reflectivity with optimum level of sensitivity.

Keywords: biosensor, sensitivity, surface plasmon resonance, transfer matrix method

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66 Resource Leveling Optimization in Construction Projects of High Voltage Substations Using Nature-Inspired Intelligent Evolutionary Algorithms

Authors: Dimitrios Ntardas, Alexandros Tzanetos, Georgios Dounias

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High Voltage Substations (HVS) are the intermediate step between production of power and successfully transmitting it to clients, making them one of the most important checkpoints in power grids. Nowadays - renewable resources and consequently distributed generation are growing fast, the construction of HVS is of high importance both in terms of quality and time completion so that new energy producers can quickly and safely intergrade in power grids. The resources needed, such as machines and workers, should be carefully allocated so that the construction of a HVS is completed on time, with the lowest possible cost (e.g. not spending additional cost that were not taken into consideration, because of project delays), but in the highest quality. In addition, there are milestones and several checkpoints to be precisely achieved during construction to ensure the cost and timeline control and to ensure that the percentage of governmental funding will be granted. The management of such a demanding project is a NP-hard problem that consists of prerequisite constraints and resource limits for each task of the project. In this work, a hybrid meta-heuristic method is implemented to solve this problem. Meta-heuristics have been proven to be quite useful when dealing with high-dimensional constraint optimization problems. Hybridization of them results in boost of their performance.

Keywords: hybrid meta-heuristic methods, substation construction, resource allocation, time-cost efficiency

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65 A Hybrid Algorithm Based on Greedy Randomized Adaptive Search Procedure and Chemical Reaction Optimization for the Vehicle Routing Problem with Hard Time Windows

Authors: Imen Boudali, Marwa Ragmoun

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The Vehicle Routing Problem with Hard Time Windows (VRPHTW) is a basic distribution management problem that models many real-world problems. The objective of the problem is to deliver a set of customers with known demands on minimum-cost vehicle routes while satisfying vehicle capacity and hard time windows for customers. In this paper, we propose to deal with our optimization problem by using a new hybrid stochastic algorithm based on two metaheuristics: Chemical Reaction Optimization (CRO) and Greedy Randomized Adaptive Search Procedure (GRASP). The first method is inspired by the natural process of chemical reactions enabling the transformation of unstable substances with excessive energy to stable ones. During this process, the molecules interact with each other through a series of elementary reactions to reach minimum energy for their existence. This property is embedded in CRO to solve the VRPHTW. In order to enhance the population diversity throughout the search process, we integrated the GRASP in our method. Simulation results on the base of Solomon’s benchmark instances show the very satisfactory performances of the proposed approach.

Keywords: Benchmark Problems, Combinatorial Optimization, Vehicle Routing Problem with Hard Time Windows, Meta-heuristics, Hybridization, GRASP, CRO

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64 Electrocatalytic Enhancement Mechanism of Dual-Atom and Single-Atom MXenes-Based Catalyst in Oxygen and Hydrogen Evolution Reactions

Authors: Xin Zhao. Xuerong Zheng. Andrey L. Rogach

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Using single metal atoms has been considered an efficient way to develop new HER and OER catalysts. MXenes, a class of two-dimensional materials, have attracted tremendous interest as promising substrates for single-atom metal catalysts. However, there is still a lack of systematic investigations on the interaction mechanisms between various MXenes substrates and single atoms. Besides, due to the poor interaction between metal atoms and substrates resulting in low loading and stability, dual-atom MXenes-based catalysts have not been successfully synthesized. We summarized the electrocatalytic enhancement mechanism of three MXenes-based single-atom catalysts through experimental and theoretical results demonstrating the stronger hybridization between Co 3d and surface-terminated O 2p orbitals, optimizing the electronic structure of Co single atoms in the composite. This, in turn, lowers the OER and HER energy barriers and accelerates the catalytic kinetics in the case of the Co@V2CTx composite. The poor interaction between single atoms and substrates can be improved by a surface modification to synthesize dual-atom catalysts. The synergistic electronic structure enhances the stability and electrocatalytic activity of the catalyst. Our study provides guidelines for designing single-atom and dual-atom MXene-based electrocatalysts and sheds light on the origins of the catalytic activity of single-atoms on MXene substrates.

Keywords: dual-atom catalyst, single-atom catalyst, MXene substrates, water splitting

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63 Perfluoroheptanoic Acid Affects Xenopus Embryo Embryogenesis by Inducing the Phosphorylation of ERK and JNK

Authors: Chowon Kim, Yoo-Kyung Kim, Kyeong Yeon Park, Hyun-Shik Lee

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Perfluoroalkyl compounds (PFCs) are globally distributed synthetic compounds that are known to adversely affect human health. Developmental toxicity assessment of PFCs is important to facilitate the evaluation of their environmental impact. In the present study, we assessed the developmental toxicity and teratogenicity of PFCs with different numbers of carbon atoms on Xenopus embryogenesis. An initial frog embryo teratogenicity assay-Xenopus (FETAX) assay was performed that identified perfluorohexanoic (PFHxA) and perfluoroheptanoic (PFHpA) acids as potential teratogens and developmental toxicants. The mechanism underlying this teratogenicity was also investigated by measuring the expression of tissue-specific biomarkers such as phosphotyrosine‑binding protein, xPTB (liver); NKX2.5 (heart); and Cyl18 (intestine). Whole‑mount in situ hybridization, reverse transcriptase‑polymerase chain reaction (RT-PCR), and histologic analyses detected severe defects in the liver and heart following exposure to PFHxA or PFHpA. In addition, immunoblotting revealed that PFHpA significantly increased the phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), while PFHxA slightly increased these, as compared with the control. These results suggest that PFHxA and PFHpA are developmental toxicants and teratogens, with PFHpA producing more severe effects on liver and heart development through the induction of ERK and JNK phosphorylation.

Keywords: PFCs, ERK, JNK, xenopus

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62 Evaluation of Real Time PCR Methods for Food Safety

Authors: Ergun Sakalar, Kubra Bilgic

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In the last decades, real-time PCR has become a reliable tool preferred to use in many laboratories for pathogen detection. This technique allows for monitoring target amplification via fluorescent molecules besides admit of quantitative analysis by enabling of convert outcomes of thermal cycling to digital data. Sensitivity and traceability of real-time PCR are based on measuring of fluorescence that appears only when fluorescent reporter dye bound to specific target DNA.The fluorescent reporter systems developed for this purpose are divided into two groups. The first group consists of intercalator fluorescence dyes such as SYBR Green, EvaGreen which binds to double-stranded DNA. On the other hand, the second group includes fluorophore-labeled oligonucleotide probes that are separated into three subgroups due to differences in mechanism of action; initial primer-probes such as Cyclicons, Angler®, Amplifluor®, LUX™, Scorpions, and the second one hydrolysis probes like TaqMan, Snake assay, finally hybridization probes, for instance, Molecular Beacons, Hybprobe/FRET, HyBeacon™, MGB-Eclipse, ResonSense®, Yin-Yang, MGB-Pleiades. In addition nucleic acid analogues, an increase of probe affinity to target site is also employed with fluorescence-labeled probes. Consequently, abundant real-time PCR detection chemistries are chosen by researcher according to the field of application, mechanism of action, advantages, and proper structures of primer/probes.

Keywords: fluorescent dye, food safety, molecular probes, nucleic acid analogues

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61 Following the Modulation of Transcriptional Activity of Genes by Chromatin Modifications during the Cell Cycle in Living Cells

Authors: Sharon Yunger, Liat Altman, Yuval Garini, Yaron Shav-Tal

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Understanding the dynamics of transcription in living cells has improved since the development of quantitative fluorescence-based imaging techniques. We established a method for following transcription from a single copy gene in living cells. A gene tagged with MS2 repeats, used for mRNA tagging, in its 3' UTR was integrated into a single genomic locus. The actively transcribing gene was detected and analyzed by fluorescence in situ hybridization (FISH) and live-cell imaging. Several cell clones were created that differed in the promoter regulating the gene. Thus, comparative analysis could be obtained without the risk of different position effects at each integration site. Cells in S/G2 phases could be detected exhibiting two adjacent transcription sites on sister chromatids. A sharp reduction in the transcription levels was observed as cells progressed along the cell cycle. We hypothesized that a change in chromatin structure acts as a general mechanism during the cell cycle leading to down-regulation in the activity of some genes. We addressed this question by treating the cells with chromatin decondensing agents. Quantifying and imaging the treated cells suggests that chromatin structure plays a role both in regulating transcriptional levels along the cell cycle, as well as in limiting an active gene from reaching its maximum transcription potential at any given time. These results contribute to understanding the role of chromatin as a regulator of gene expression.

Keywords: cell cycle, living cells, nucleus, transcription

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60 Investigation of Rifampicin and Isoniazid Resistance Mutated Genes in Mycobacterium Tuberculosis Isolated From Patients

Authors: Seyyed Mohammad Amin Mousavi Sagharchi, Alireza Mahmoudi Nasab, Tim Bakker

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Introduction: Mycobacterium tuberculosis (MTB) is the most intelligent bacterium that existed in the world to our best knowledge. This bacterium can cause tuberculosis (TB) which is responsible for its spread speed and murder of millions of people around the world. MTB has the practical function to escape from anti-tuberculosis drugs (AT), for this purpose, it handles some mutations in the main genes and creates new patterns for inhibited genes. Method and materials: Researchers have their best tries to safely isolate MTB from the sputum specimens of 35 patients in some hospitals in the Tehran province and detect MTB by culture on Löwenstein-Jensen (LJ) medium and microscopic examination. DNA was extracted from the established bacterial colony by enzymatic extraction method. It was amplified by the polymerase chain reaction (PCR) method, reverse hybridization, and evaluation for detection of resistance genes; generally, researchers apply GenoType MTBDRplus assay. Results: Investigations of results declare us that 21 of the isolated specimens (about 60%) have mutation in rpoB gene, which resisted to rifampicin (most prevalence), and 8 of them (about 22.8%) have mutation in katG or inhA genes which resisted to isoniazid. Also, 4 of them (about 11.4%) don't have any mutation, and 2 of them (about 5.7%) have mutation in every three genes, which makes them resistant to the two drugs mentioned above. Conclusion: Rifampicin and isoniazid are two essential AT that using in the first line of treatment. Resistance in rpoB, and katG, and inhA genes related to mentioned drugs lead to ineffective treatment.

Keywords: mycobacterium tuberculosis, tuberculosis, drug resistance, isoniazid, rifampicin

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59 Heritability and Diversity Analysis of Blast Resistant Upland Rice Genotypes Based on Quantitative Traits

Authors: Mst. Tuhina-Khatun, Mohamed Hanafi Musa, Mohd Rafii Yosup, Wong Mui Yun, Md. Aktar-Uz-Zaman, Mahbod Sahebi

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Rice is a staple crop of economic importance of most Asian people, and blast is the major constraints for its higher yield. Heritability of plants traits helps plant breeders to make an appropriate selection and to assess the magnitude of genetic improvement through hybridization. Diversity of crop plants is necessary to manage the continuing genetic erosion and address the issues of genetic conservation for successfully meet the future food requirements. Therefore, an experiment was conducted to estimate heritability and to determine the diversity of 27 blast resistant upland rice genotypes based on 18 quantitative traits using randomized complete block design. Heritability value was found to vary from 38 to 93%. The lowest heritability belonged to the character total number of tillers/plant (38%). In contrast, number of filled grains/panicle, and yield/plant (g) was recorded for their highest heritability value viz. 93 and 91% correspondingly. Cluster analysis based on 18 traits grouped 27 rice genotypes into six clusters. Cluster I was the biggest, which comprised 17 genotypes, accounted for about 62.96% of total population. The multivariate analysis suggested that the genotype ‘Chokoto 14’ could be hybridized with ‘IR 5533-55-1-11’ and ‘IR 5533-PP 854-1’ for broadening the gene pool of blast resistant upland rice germplasms for yield and other favorable characters.

Keywords: blast resistant, diversity analysis, heritability, upland rice

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58 Development of an Ecological Binder by Geopolymerization of Untreated Dredged Sediments

Authors: Lisa Monteiro, Jacqueline Saliba, Nadia Saiyouri, Humberto Y. Godoy

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Theevolution of the global environmental context incites companies to reduce their impact by reusing local materials and promoting circular economy. Dredged sediments represent a potential source of materials due to their large volume. Indeed, the dredging operations carried out in Gironde alone generated an annual volume of sediment of approximately 9 million m³. Moreover, on the eve of the evolution of laws concerning dredging practices, the recovery of sediments is necessary to create a viable economy for their management. This thesis work is oriented towards the development of an ecological binder from the fine fraction of untreated dredged sediments. In fact, their physico-chemical properties make them favorable for the synthesis of geopolymer, current competitor of cement, thanks to its lower carbon footprint and environmental impact. However, several obstacles must be overcome before implementing this new family of materials: the use of sediments without thermal or chemical treatment, the absence of a formulation approach, ignorance of the reactions produced, etc. During the first year of the thesis, a physico-chemical characterization of the sediments made it possible to validate their use as precursors forgeopolymerization according to three criteria: their fineness, their mineralogical composition, and the percentage of amorphous phase. Following these results, several formulations have been defined, taking into account the environmental impact. The sediments were activated with an alkaline solution of sodium hydroxide and sodium silicate. Two other formulations with cement and blast furnace slag have been defined for comparison. The results highlighted the possibility of forming geopolymers from untreated and still wet dredged sediments. The development of structural bonds through the formation of hydrated sodium aluminosilicate thus leads to higher strengths at 90 days (4.78 MPa) than a mixture with cement (0.75 MPa). A 30% gain in CO₂ emissions has also been obtained compared to cement. In order to reduce the uncertainties linked to the absence of a formulation approach, to optimize the number of experiments to be carried out in the laboratory, and to obtain an optimal formulation, an analysis by mixing plan was conducted in order to frame the responses according to the proportions of the constituents. Following the obtaining of an optimal binder, the work will focus on the study of the durability and the interspecific variability of the sediments on the mechanical properties by testing the binder developed with different sediments dredged from the Bordeaux estuary. , the Grand Port Maritime of Bayonne, La Rochelle, and the Bassinsd'Arcachon.

Keywords: compressive strength, dredged sediments, ecological binder, geopolymers

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57 An Evaluation of Renewable Energy Sources in Green Building Systems for the Residential Sector in the Metropolis, Kolkata, India

Authors: Tirthankar Chakraborty, Indranil Mukherjee

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The environmental aspect had a major effect on industrial decisions after the deteriorating condition of our surroundings dsince the industrial activities became apparent. Green buildings have been seen as a possible solution to reduce the carbon emissions from construction projects and the housing industry in general. Though this has been established in several areas, with many commercial buildings being designed green, the scope for expansion is still significant and further information on the importance and advantages of green buildings is necessary. Several commercial green building projects have come up and the green buildings are mainly implemented in the residential sector when the residential projects are constructed to furnish amenities to a large population. But, residential buildings, even those of medium sizes, can be designed to incorporate elements of sustainable design. In this context, this paper attempts to give a theoretical appraisal of the use of renewable energy systems in residential buildings of different sizes considering the weather conditions (solar insolation and wind speed) of the metropolis, Kolkata, India. Three cases are taken; one with solar power, one with wind power and one with a combination of the two. All the cases are considered in conjunction with conventional energy, and the efficiency of each in fulfilling the total energy demand is verified. The optimum combination for reducing the carbon footprint of the residential building is thus established. In addition, an assessment of the amount of money saved due to green buildings in metered water supply and price of coal is also mentioned.

Keywords: renewable energy, green buildings, solar power, wind power, energy hybridization, residential sector

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56 Mechanical and Optical Properties of Doped Aluminum Nitride Thin Films

Authors: Padmalochan Panda, R. Ramaseshan

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Aluminum nitride (AlN) is a potential candidate for semiconductor industry due to its wide band gap (6.2 eV), high thermal conductivity and low thermal coefficient of expansion. A-plane oriented AlN film finds an important role in deep UV-LED with higher isotropic light extraction efficiency. Also, Cr-doped AlN films exhibit dilute magnetic semiconductor property with high Curie temperature (300 K), and thus compatible with modern day microelectronics. In this work, highly a-axis oriented wurtzite AlN and Al1-xMxN (M = Cr, Ti) films have synthesized by reactive co-sputtering technique at different concentration. Crystal structure of these films is studied by Grazing incidence X-ray diffraction (GIXRD) and Transmission electron microscopy (TEM). Identification of binding energy and concentration (x) in these films is carried out by X-ray photoelectron spectroscopy (XPS). Local crystal structure around the Cr and Ti atom of these films are investigated by X-ray absorption spectroscopy (XAS). It is found that Cr and Ti replace the Al atom in AlN lattice and the bond lengths in first and second coordination sphere with N and Al, respectively, decrease concerning doping concentration due to strong p-d hybridization. The nano-indentation hardness of Cr and Ti-doped AlN films seems to increase from 17.5 GPa (AlN) to around 23 and 27.5 GPa, respectively. An-isotropic optical properties of these films are studied by the Spectroscopic Ellipsometry technique. Refractive index and extinction coefficient of these films are enhanced in normal dispersion region as compared to the parent AlN film. The optical band gap energies also seem to vary between deep UV to UV regions with the addition of Cr, thus by bringing out the usefulness of these films in the area of optoelectronic device applications.

Keywords: ellipsometry, GIXRD, hardness, XAS

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55 Evaluation of Two Earliness Cotton Genotypes in Three Ecological Regions

Authors: Gholamhossein Hosseini

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Two earliness cotton genotypes I and II, which had been developed by hybridization and backcross methods between sindise-80 as an early maturing gene parent and two other lines i.e. Red leaf and Bulgare-557 as a second parent, are subjected to different environmental conditions. The early maturing genotypes with coded names of I and II were compared with four native cotton cultivars in randomized complete block design (RCBD) with four replications in three ecological regions of Iran from 2016-2017. Two early maturing genotypes along with four native cultivars viz. Varamin, Oltan, Sahel and Arya were planted in Agricultural Research Station of Varamin, Moghan and Kashmar for evaluation. Earliness data were collected for six treatments during two years in the three regions except missing data for the second year of Kashmar. Therefore, missed data were estimated and imputed. For testing the homogeneity of error variances, each experiment at a given location or year is analyzed separately using Hartley and Bartlett’s Chi-square tests and both tests confirmed homogeneity of variance. Combined analysis of variance showed that genotypes I and II were superior in Varamin, Moghan and Kashmar regions. Earliness means and their interaction effects were compared with Duncan’s multiple range tests. Finally combined analysis of variance showed that genotypes I and II were superior in Varamin, Moghan and Kashmar regions. Earliness means and their interaction effects are compared with Duncan’s multiple range tests.

Keywords: cotton, combined, analysis, earliness

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54 YHV-Responsive Gene Expression under the Influence of PmRelish Regulation

Authors: Suwattana Visetnan, Premruethai Supungul, Sureerat Tang, Ikuo Hirono, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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In animals, infection by Gram-negative bacteria and certain viruses activates the Imd signaling pathway wherein the a NF-κB transcription factor, Relish, is a key regulatory protein for the synthesis of antimicrobial proteins. Infection by yellow head virus (YHV) activates the Imd pathway. To investigate the expression of genes involved in YHV infection and under the influence of PmRelish regulation, RNA interference and suppression subtractive hybridization (SSH) are employed. The genes in forward library expressed in shrimp after YHV infection and under the activity of PmRelish were obtained by subtracting the cDNAs from YHV-infected and PmRelish-knockdown shrimp with cDNAs from YHV-infected shrimp. Opposite subtraction gave a reverse library whereby an alternative set of genes under YHV infection and no PmRelish expression was obtained. Sequencing of 252 and 99 cDNA clones from the respective forward and reverse libraries were done and annotated through blast search against the GenBank sequences. Genes involved in defense and homeostasis were abundant in both libraries, 31% and 23% in the forward and reverse libraries, respectively. They were predominantly antimicrobial proteins, proteinases and proteinase inhibitors. The expression of antimicrobial protein genes, ALFPm3, crustinPm1, penaeidin3 and penaeidin5 were tested under PmRelish silencing and Gram-negative bacterium V. harveyi infection. Together with the results previously reported, the expression of penaeidin5 and also penaeidin3 but not ALFPm3 and crustinPm1 were under the regulation of PmRelish in the Imd pathway.

Keywords: relish, yellow head virus, penaeus monodon, antimicrobial proteins

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53 Evaluation of Developmental Toxicity and Teratogenicity of Perfluoroalkyl Compounds Using FETAX

Authors: Hyun-Kyung Lee, Jehyung Oh, Young Eun Jeong, Hyun-Shik Lee

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Perfluoroalkyl compounds (PFCs) are environmental toxicants that persistently accumulate in the human blood. Their widespread detection and accumulation in the environment raise concerns about whether these chemicals might be developmental toxicants and teratogens in the ecosystem. We evaluated and compared the toxicity of PFCs of containing various numbers of carbon atoms (C8-11 carbons) on vertebrate embryogenesis. We assessed the developmental toxicity and teratogenicity of various PFCs. The toxic effects on Xenopus embryos were evaluated using different methods. We measured teratogenic indices (TIs) and investigated the mechanisms underlying developmental toxicity and teratogenicity by measuring the expression of organ-specific biomarkers such as xPTB (liver), Nkx2.5 (heart), and Cyl18 (intestine). All PFCs that we tested were found to be developmental toxicants and teratogens. Their toxic effects were strengthened with increasing length of the fluorinated carbon chain. Furthermore, we produced evidence showing that perfluorodecanoic acid (PFDA) and perfluoroundecanoic acid (PFuDA) are more potent developmental toxicants and teratogens in an animal model compared to the other PFCs we evaluated [perfluorooctanoic acid (PFOA) and perfluorononanoic acid (PFNA)]. In particular, severe defects resulting from PFDA and PFuDA exposure were observed in the liver and heart, respectively, using the whole mount in situ hybridization, real-time PCR, pathologic analysis of the heart, and dissection of the liver. Our studies suggest that most PFCs are developmental toxicants and teratogens, however, compounds that have higher numbers of carbons (i.e., PFDA and PFuDA) exert more potent effects.

Keywords: PFC, xenopus, fetax, development

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52 Thin Films of Glassy Carbon Prepared by Cluster Deposition

Authors: Hatem Diaf, Patrice Melinon, Antonio Pereira, Bernard Moine, Nicholas Blanchard, Florent Bourquard, Florence Garrelie, Christophe Donnet

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Glassy carbon exhibits excellent biological compatibility with live tissues meaning it has high potential for applications in life science. Moreover, glassy carbon has interesting properties including 'high temperature resistance', hardness, low density, low electrical resistance, low friction, and low thermal resistance. The structure of glassy carbon has long been a subject of debate. It is now admitted that glassy carbon is 100% sp2. This term is a little bit confusing as long sp2 hybridization defined from quantum chemistry is related to both properties: threefold configuration and pi bonding (parallel pz orbitals). Using plasma laser deposition of carbon clusters combined with pulsed nano/femto laser annealing, we are able to synthesize thin films of glassy carbon of good quality (probed by G band/ D disorder band ratio in Raman spectroscopy) without thermal post annealing. A careful inspecting of Raman signal, plasmon losses and structure performed by HRTEM (High Resolution Transmission Electron Microscopy) reveals that both properties (threefold and pi orbitals) cannot coexist together. The structure of the films is compared to models including schwarzites based from negatively curved surfaces at the opposite of onions or fullerene-like structures with positively curved surfaces. This study shows that a huge collection of porous carbon named vitreous carbon with different structures can coexist.

Keywords: glassy carbon, cluster deposition, coating, electronic structure

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51 White Light Emitting Carbon Dots- Surface Modification of Carbon Dots Using Auxochromes

Authors: Manasa Perikala, Asha Bhardwaj

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Fluorescent carbon dots (CDs), a young member of Carbon nanomaterial family, has gained a lot of research attention across the globe due to its highly luminescent emission properties, non-toxic behavior, stable emission properties, and zero re-absorption lose. These dots have the potential to replace the use of traditional semiconductor quantum dots in light-emitting devices (LED’s, fiber lasers) and other photonic devices (temperature sensor, UV detector). However, One major drawback of Carbon dots is that, till date, the actual mechanism of photoluminescence (PL) in carbon dots is still an open topic of discussion among various researchers across the globe. PL mechanism of CDs based on wide particle size distribution, the effect of surface groups, hybridization in carbon, and charge transfer mechanisms have been proposed. Although these mechanisms explain PL of CDs to an extent, no universally accepted mechanism to explain complete PL behavior of these dots is put forth. In our work, we report parameters affecting the size and surface of CDs, such as time of the reaction, synthesis temperature and concentration of precursors and their effects on the optical properties of the carbon dots. The effect of auxochromes on the emission properties and re-modification of carbon surface using an external surface functionalizing agent is discussed in detail. All the explanations have been supported by UV-Visible absorption, emission spectroscopies, Fourier transform infrared spectroscopy and Transmission electron microscopy and X-Ray diffraction techniques. Once the origin of PL in CDs is understood, parameters affecting PL centers can be modified to tailor the optical properties of these dots, which can enhance their applications in the fabrication of LED’s and other photonic devices out of these carbon dots.

Keywords: carbon dots, photoluminescence, size effects on emission in CDs, surface modification of carbon dots

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50 Ending Communal Conflicts in Africa: The Relevance of Traditional Approaches to Conflict Resolution

Authors: Kindeye Fenta Mekonnen, Alagaw Ababu Kifle

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The failure of international responses to armed conflict to address local preconditions for national stability has recently attracted what has been called the ‘local turn’ in peace building. This ‘local turn’ in peace building amplified a renewed interest in traditional/indigenous methods of conflict resolution, a field that has been hitherto dominated by anthropologists with their focus on the procedures and rituals of such approaches. This notwithstanding, there is still limited empirical work on the relevance of traditional methods of conflict resolution to end localized conflicts vis-à-vis hybrid and modern approaches. The few exceptions to this generally draw their conclusion from very few (almost all successful) cases that make it difficult to judge the validity and cross-case application of their results. This paper seeks to fill these gaps by undertaking a quantitative analysis of the trend and applications of different communal conflict resolution initiatives, their potential to usher in long-term peace, and the extent to which their outcomes are influenced by the intensity and scope of a conflict. The paper makes the following three tentative conclusions. First, traditional mechanisms and traditional actors still dominate the communal conflict resolution landscape, either individually or in combination with other methods. Second, traditional mechanisms of conflict resolution tend to be more successful in ending a conflict and preventing its re-occurrence compared to hybrid and modern arrangements. This notwithstanding and probably due to the scholarly call for local turn in peace building, contemporary communal conflict resolution approaches are becoming less and less reliant on traditional mechanisms alone and (therefore) less effective. Third, there is yet inconclusive evidence on whether hybridization is an asset or a liability in the resolution of communal conflicts and the extent to which this might be mediated by the intensity of a conflict.

Keywords: traditional conflict resolution, hybrid conflict resolution, communal conflict, relevance, conflict intensity

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49 Geographical Parthenogenesis in Plants

Authors: Elvira Hörandl

Abstract:

The term “Geographical parthenogenesis” describes the phenomenon that asexual organisms usually occupy larger and more northern distribution areas than their sexual relatives and tend to colonize previously glaciated areas. Several case studies in flowering plants confirm the geographical pattern, but the causal factors behind the phenomenon are still unclear. Previous authors regarded predominant polyploidy in asexual (apomictic) plants as the main factor. However, the geographical pattern is not the rule for sexual polyploids. Recent research confirmed a previous hypothesis of the author that a combination of factors is acting: Although uniparental reproduction provides better colonization abilities, it is most efficient in combination with polyploidy. I will present results on case studies in the genus Ranunculus of both autopolyploid and allopolyploid species and species complexes reproducing via facultative apomixis. Polyploidy seems to contribute mainly to a better tolerance of colder climates and temperate extremes, whereby epigenetic flexibility, changes in gene expression, and phenotypic plasticity play an important role in occupying ecological niches under harsh conditions. Phylogenomic studies entangle complex hybrid origins of asexual taxa, which increases intragenomic heterozygosity of asexual plants. Interestingly, our results suggest an association of sexuality with abiotic stresses, specifically with light stress, which might explain that still, most plants in high altitudes and in southern areas retain sexual reproduction despite other climatic conditions that would favor apomictic plants. We conclude that geographical parthenogenesis results from the complex interplay of the genomic constitution, mode of reproduction and environmental factors.

Keywords: apomixis, polyploidy, hybridization, abiotic stress, epigenetics, phylogenomics

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48 Genomic and Evolutionary Diversity of Long Terminal Repeat (LTR) Retrotransposons in Date Palm (Phoenix dactylifera)

Authors: Faisal Nouroz, Mukaramin Mukaramin

Abstract:

Of the transposable elements (TEs), the retrotransposons are the most copious elements identified from many sequenced genomes. They have played a major role in genome evolution, rearrangement, and expansions based on their copy and paste mode of proliferation. They are further divided into LTR and Non-LTR retrotransposons. The purpose of the current study was to identify the LTR REs in sequenced Phoenix dactylifera genome and to study their structural diversity. A total of 150 P. dactylifera BAC sequences with > 60kb sizes were randomly retrieved from National Center for Biotechnology Information (NCBI) database and screened for the presence of LTR retrotransposons. Seven bacterial artificial chromosomes (BAC) sequences showed full-length LTR Retrotransposons with 4 Copia and 3 Gypsy families having variable copy numbers in respective families. Reverse transcriptase (RT) domain was found as the most conserved domain among Copia and Gypsy superfamilies and was used to deduce evolutionary analysis. The amino acid residues among various RT sequences showed variability in their percentages indicating post divergence evolution. Amino acid Leucine was found in highest proportions followed by Lysine, while Methionine and Tryptophan were in lowest percentages. The phylogenetic analysis based on RT domains confirmed that although having most conserved RT regions, several evolutionary events occurred causing nucleotide polymorphisms and hence clustering of Gypsy and Copia superfamilies into their respective lineages. The study will be helpful in identification and annotation of these elements in other species and genera and their distribution patterns on chromosomes by fluorescent in situ hybridization techniques.

Keywords: transposable elements, Phoenix dactylifera, retrotransposons, phylogenetic analysis

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47 Quantum Dot – DNA Conjugates for Biological Applications

Authors: A. Banerjee, C. Grazon, B. Nadal, T. Pons, Y. Krishnan, B. Dubertret

Abstract:

Quantum Dots (QDs) have emerged as novel fluorescent probes for biomedical applications. The photophysical properties of QDs such as broad absorption, narrow emission spectrum, reduced blinking, and enhanced photostability make them advantageous over organic fluorophores. However, for some biological applications, QDs need to be first targeted to specific intracellular locations. It parallel, base pairing properties and biocompatibility of DNA has been extensively used for biosensing, targetting and intracellular delivery of numerous bioactive agents. The combination of the photophysical properties of QDs and targettability of DNA has yielded fluorescent, stable and targetable nanosensors. QD-DNA conjugates have used in drug delivery, siRNA, intracellular pH sensing and several other applications; and continue to be an active area of research. In this project, a novel method to synthesise QD-DNA conjugates and their applications in bioimaging are investigated. QDs are first solubilized in water using a thiol based amphiphilic co-polymer and, then conjugated to amine functionalized DNA using a heterobifunctional linker. The conjugates are purified by size exclusion chromatography and characterized by UV-Vis absorption and fluorescence spectroscopy, electrophoresis and microscopy. Parameters that influence the conjugation yield such as reducing agents, the excess of salt and pH have been investigated in detail. In optimized reaction conditions, up to 12 single-stranded DNA (15 mer length) can be conjugated per QD. After conjugation, the QDs retain their colloidal stability and high quantum yield; and the DNA is available for hybridization. The reaction has also been successfully tested on QDs emitting different colors and on Gold nanoparticles and therefore highly generalizable. After extensive characterization and robust synthesis of QD-DNA conjugates in vitro, the physical properties of these conjugates in cellular milieu are being invistigated. Modification of QD surface with DNA appears to remarkably alter the fate of QD inside cells and can have potential implications in therapeutic applications.

Keywords: bioimaging, cellular targeting, drug delivery, photostability

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46 MNH-886(Bt.): A Cotton Cultivar (G. Hirsutum L.) for Cultivation in Virus Infested Regions of Pakistan, Having High Seed Cotton Yield and Desirable Fibre Characteristics

Authors: Wajad Nazeer, Saghir Ahmad, Khalid Mahmood, Altaf Hussain, Abid Mahmood, Baoliang Zhou

Abstract:

MNH-886(Bt.) is a upland cotton cultivar (Gossypium hirsutum L.) developed through hybridization of three parents [(FH-207×MNH-770)×Bollgard-1] at Cotton Research Station Multan, Pakistan. It is resistant to CLCuVD with 16.25 % disease incidence (60 DAS, March sowing) whereas moderately susceptible to CLCuVD when planted in June with disease incidence 34 % (60 DAS). This disease reaction was lowest among 25 cotton advanced lines/varieties tested at hot spots of CLCuVD. Its performance was tested during 2009 to 2012 in various indigenous, provincial, and national varietal trials in comparison with the commercial variety IR-3701 and AA-802 & CIM-496. In PCCT trial during 2009-10; 2011-12, MNH-886 surpassed all the existing Bt. strains along with commercial varieties across the Punjab province with seed cotton yield production 2658 kg ha-1 and 2848 kg ha-1 which was 81.31 and 13% higher than checks, respectively. In National Coordinated Bt. Trial, MNH-886(Bt.) produced 3347 kg ha-1 seed cotton at CCRI, Multan; the hot spot of CLCuVD, in comparison to IR-3701 which gave 2556 kg ha-1. It possesses higher lint percentage (41.01%), along with the most desirable fibre traits (staple length 28.210mm, micronaire value 4.95 µg inch-1 and fibre strength 99.5 tppsi, and uniformity ratio 82.0%). The quantification of toxicity level of crystal protein was found positive for Cry1Ab/Ac protein with toxicity level 2.76µg g-1 and Mon 531 event was confirmed. Having tremendous yield potential, good fibre traits, and great tolerance to CLCuVD we can recommended this variety for cultivation in CLCuVD hotspots of Pakistan.

Keywords: cotton, cultivar, cotton leaf curl virus, CLCuVD hit districts

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45 MiR-200a/ZEB1 Pathway in Liver Fibrogenesis of Biliary Atresia

Authors: Hai-Ying Liu, Yi-Hao Chen, Shu-Yin Pang, Feng-Hua Wang, Xiao-Fang Peng, Li-Yuan Yang, Zheng-Rong Chen, Yi Chen, Bing Zhu

Abstract:

Objective: Biliary atresia (BA) is characterized by progressive liver fibrosis. Epithelial-mesenchymal transition (EMT) has been implicated as a key mechanism in the pathogenesis of organ fibrosis. MiR-200a has been shown to repress EMT. We aim to explore the role of miR-200a in the fibrogenesis of BA. Methods: We obtained the plasma samples and liver samples from patients with BA or controls to examine the role of miR-200a. Histological liver fibrosis was assessed using the Ishak fibrosis scores. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed to detect the expression of miR-200a in plasma. We also evaluated the expression of miR-200a in liver tissues using tyramide signal amplification fluorescence in situ hybridization (TSA-FISH). The expression of EMT related proteins zinc finger E-box-binding homeobox 1 (ZEB1), E-cadherin and α-smooth muscle actin (α-SMA) in the liver sections were detected by immunohistochemical staining. Results: We found that the expression of miR-200a was both elevated in the plasma and liver tissues from BA patients compared with the controls. The hepatic expression of ZEB1 and α-SMA were markedly increased in the liver sections from BA patients compared to the controls, whereas E-cadherin was downregulated in the BA group. Simultaneously, we noted that the hepatic expression of miR-200a, E-cadherin and α-SMA were upregulated with the progression of liver fibrosis in the BA group, while ZEB1 was downregulated with the progression of liver fibrosis in BA patients. Conclusion: These findings suggest EMT has a critical effect on the fibrotic process of BA, and the interaction between miR-200a and ZEB1 may regulate EMT and eventually influence liver fibrogenesis of BA.

Keywords: biliary atresia, liver fibrosis, MicroRNA, epithelial-mesenchymal transition, zinc finger E-box-binding homeobox 1

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44 Phylogenetic Relationships between the Whole Sets of Individual Flow Sorted U, M, S and C Chromosomes of Aegilops and Wheat as Revealed by COS Markers

Authors: András Farkas, István Molnár, Jan Vrána, Veronika Burešová, Petr Cápal, András Cseh, Márta Molnár-Láng, Jaroslav Doležel

Abstract:

Species of Aegilops played a central role in the evolution of wheat and are sources of traits related to yield quality and tolerance against biotic and abiotic stresses. These wild genes and alleles are desirable to use in crop improvement programs via introgressive hybridization. However, the success of chromosome mediated gene transfer to wheat are hampered by the pour knowledge on the genome structure of Aegilops relative to wheat and by the low number of cost-effective molecular markers specific for Aegilops chromosomes. The COS markers specific for genes conserved throughout evolution in both sequence and copy number between Triticeae/Aegilops taxa and define orthologous regions, thus enabling the comparison of regions on the chromosomes of related species. The present study compared individual chromosomes of Aegilops umbellulata (UU), Ae. comosa (MM), Ae. speltoides (SS) and Ae. caudata (CC) purified by flourescent labelling with oligonucleotid SSR repeats and biparametric flow cytometry with wheat by identifying orthologous chromosomal regions by COS markers. The linear order of bin-mapped COS markers along the wheat D chromosomes was identified by the use of chromosome-specific sequence data and virtual gene order. Syntenic regions of wheat identifying genome rearrangements differentiating the U, M, S or C genomes from the D genome of wheat were detected. The conserved orthologous set markers assigned to Aegilops chromosomes promise to accelerate gene introgression by facilitating the identification of alien chromatin. The syntenic relationships between the Aegilops species and wheat will facilitate the targeted development of new markers specific for U, M, S and C genomic regions and will contribute to the understanding of molecular processes related to the evolution of Aegilops.

Keywords: Aegilops, cos-markers, flow-sorting, wheat

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43 Salt Tolerance of Potato: Genetically Engineered with Atriplex canescens BADH Gene Driven by 3 Copies of CAMV35s Promoter

Authors: Arfan Ali, Muhammad Shahzad Iqbal, Idrees Ahmad Nasir

Abstract:

Potato (Solanum tuberosum L.) is ranked among the top leading staple foods in the world. Salinity adversely affects potato crop yield and quality. Therefore, increased level of salt tolerance is a key factor to ensure high yield. The present study focused on the Agrobacterium-mediated transformation of Atriplex canescens betaine aldehyde dehydrogenase (BADH) gene, using single, double and triple CAMV35s promoter to improve salt tolerance in potato. Detection of seven potato lines harboring BADH gene, followed by identification of T-DNA insertions, determination of transgenes copies no through Southern Hybridization and quantification of BADH protein through Enzyme Linked Immunosorbent Assay were considered in this study. The results clearly depict that the salt tolerance of potato was found to be promoter-dependent, as the potato transgenic lines with triple promoter showed 4.4 times more glycine betaine production which consequently leads towards high resistance to salt stress as compared to transgenic potato lines with single and double promoters having least production of glycine betaine. Moreover, triple promoter transgenic potato lines have also shown lower levels of H2O2, malondialdehyde (MDA), relative electrical conductivity, high proline and chlorophyll content as compared other two lines having a single and double promoter. Insilco analysis also confirmed that Atriplex canescens BADH has the tendency to interact with sodium ions and water molecules. Taken together these facts it can be concluded that over-expression of BADH under triple CAMV35s promoter with more glycine betaine, chlorophyll & MDA contents, high relative quantities of other metabolites results in an enhanced level of salt tolerance in potato.

Keywords: Atriplex canescens, BADH, CAMV35s promotor, potato, Solanum tubersum

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42 Human Facial Emotion: A Comparative and Evolutionary Perspective Using a Canine Model

Authors: Catia Correia Caeiro, Kun Guo, Daniel Mills

Abstract:

Despite its growing interest, emotions are still an understudied cognitive process and their origins are currently the focus of much debate among the scientific community. The use of facial expressions as traditional hallmarks of discrete and holistic emotions created a circular reasoning due to a priori assumptions of meaning and its associated appearance-biases. Ekman and colleagues solved this problem and laid the foundations for the quantitative and systematic study of facial expressions in humans by developing an anatomically-based system (independent from meaning) to measure facial behaviour, the Facial Action Coding System (FACS). One way of investigating emotion cognition processes is by applying comparative psychology methodologies and looking at either closely-related species (e.g. chimpanzees) or phylogenetically distant species sharing similar present adaptation problems (analogy). In this study, the domestic dog was used as a comparative animal model to look at facial expressions in social interactions in parallel with human facial expressions. The orofacial musculature seems to be relatively well conserved across mammal species and the same holds true for the domestic dog. Furthermore, the dog is unique in having shared the same social environment as humans for more than 10,000 years, facing similar challenges and acquiring a unique set of socio-cognitive skills in the process. In this study, the spontaneous facial movements of humans and dogs were compared when interacting with hetero- and conspecifics as well as in solitary contexts. In total, 200 participants were examined with FACS and DogFACS (The Dog Facial Action Coding System): coding tools across four different emotionally-driven contexts: a) Happiness (play and reunion), b) anticipation (of positive reward), c) fear (object or situation triggered), and d) frustration (negation of a resource). A neutral control was added for both species. All four contexts are commonly encountered by humans and dogs, are comparable between species and seem to give rise to emotions from homologous brain systems. The videos used in the study were extracted from public databases (e.g. Youtube) or published scientific databases (e.g. AM-FED). The results obtained allowed us to delineate clear similarities and differences on the flexibility of the facial musculature in the two species. More importantly, they shed light on what common facial movements are a product of the emotion linked contexts (the ones appearing in both species) and which are characteristic of the species, revealing an important clue for the debate on the origin of emotions. Additionally, we were able to examine movements that might have emerged for interspecific communication. Finally, our results are discussed from an evolutionary perspective adding to the recent line of work that supports an ancient shared origin of emotions in a mammal ancestor and defining emotions as mechanisms with a clear adaptive purpose essential on numerous situations, ranging from maintenance of social bonds to fitness and survival modulators.

Keywords: comparative and evolutionary psychology, emotion, facial expressions, FACS

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41 Obtaining Triploid Plants of Sprekelia formosissima by Artificial Hybridization

Authors: Jose Manuel Rodriguez-Dominguez, Rodrigo Barba-Gonzalez, Ernesto Tapia-Campos

Abstract:

Sprekelia formosissima (L.) Herbert is a bulbous ornamental species of the monocotyledonous Amaryllidaceae family, and it is a perennial, herbaceous monotypic plant commonly known as ‘Aztec Lily’ or ‘Jacobean Lily’; it is distributed through Mexico and Guatemala. Its scarlet flowers with curved petals have made it an exceptional ornamental pot plant. Cytogenetic studies in this species have shown differences in chromosome number (2n=60, 120, 150, 180) with a basic number x=30. Different reports have shown a variable ploidy level (diploid, tetraploid, pentaploid and hexaploid); however, triploid plants have not been reported. In this work, triploid plants of S. formosissima were obtained by crossing tetraploid (2n=4x=120) with diploid (2n=2x=60) genotypes of this species; the seeds obtained from the crosses were placed in pots with a moist substrate made of Peat Moss: Vermiculite (7:3) for germination. Root tips were collected, and metaphasic chromosome preparations were performed. For chromosome counting, the best five metaphases obtained were photographed with a Leica DMRA2 microscope (Leica Microsystems, Germany) microscopy coupled to an Evolution QEI camera under phase contrast (Media-Cybernetics). Chromosomes counting in root-tip cells showed that 100% of the plants were triploid (2n=3x=90). Although tetraploid or pentaploid plants of S. formosissima are highly appreciated, they usually have lower growth rates than related diploid ones. For this reason, it is important to obtain triploid plants, which have advantages such as higher growth rates than tetraploid and pentaploid, larger flowers than those of the diploid plants and they are expected to not be able to produce seeds because their gametes are aneuploids. Furthermore, triploids may become very important for genomic research in the future, creating opportunities for discovering and monitoring genomic and transcriptomic changes in unbalanced genomes, hence the importance of this work.

Keywords: Amaryllidaceae, cytogenetics, ornamental, ploidy level

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40 DNA Barcoding for Identification of Dengue Vectors from Assam and Arunachal Pradesh: North-Eastern States in India

Authors: Monika Soni, Shovonlal Bhowmick, Chandra Bhattacharya, Jitendra Sharma, Prafulla Dutta, Jagadish Mahanta

Abstract:

Aedes aegypti and Aedes albopictus are considered as two major vectors to transmit dengue virus. In North-east India, two states viz. Assam and Arunachal Pradesh are known to be high endemic zone for dengue and Chikungunya viral infection. The taxonomical classification of medically important vectors are important for mapping of actual evolutionary trends and epidemiological studies. However, misidentification of mosquito species in field-collected mosquito specimens could have a negative impact which may affect vector-borne disease control policy. DNA barcoding is a prominent method to record available species, differentiate from new addition and change of population structure. In this study, a combined approach of a morphological and molecular technique of DNA barcoding was adopted to explore sequence variation in mitochondrial cytochrome c oxidase subunit I (COI) gene within dengue vectors. The study has revealed the map distribution of the dengue vector from two states i.e. Assam and Arunachal Pradesh, India. Approximate five hundred mosquito specimens were collected from different parts of two states, and their morphological features were compared with the taxonomic keys. The analysis of detailed taxonomic study revealed identification of two species Aedes aegypti and Aedes albopictus. The species aegypti comprised of 66.6% of the specimen and represented as dominant dengue vector species. The sequences obtained through standard DNA barcoding protocol were compared with public databases, viz. GenBank and BOLD. The sequences of all Aedes albopictus have shown 100% similarity whereas sequence of Aedes aegypti has shown 99.77 - 100% similarity of COI gene with that of different geographically located same species based on BOLD database search. From dengue prevalent different geographical regions fifty-nine sequences were retrieved from NCBI and BOLD databases of the same and related taxa to determine the evolutionary distance model based on the phylogenetic analysis. Neighbor-Joining (NJ) and Maximum Likelihood (ML) phylogenetic tree was constructed in MEGA6.06 software with 1000 bootstrap replicates using Kimura-2-Parameter model. Data were analyzed for sequence divergence and found that intraspecific divergence ranged from 0.0 to 2.0% and interspecific divergence ranged from 11.0 to 12.0%. The transitional and transversional substitutions were tested individually. The sequences were deposited in NCBI: GenBank database. This observation claimed the first DNA barcoding analysis of Aedes mosquitoes from North-eastern states in India and also confirmed the range expansion of two important mosquito species. Overall, this study insight into the molecular ecology of the dengue vectors from North-eastern India which will enhance the understanding to improve the existing entomological surveillance and vector incrimination program.

Keywords: COI, dengue vectors, DNA barcoding, molecular identification, North-east India, phylogenetics

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