Search results for: site directed mutagenesis

Authors: Yakup Ulusu, Numan Eczacıoğlu, İsa Gökçe, Helen Waller, Jeremy H. Lakey

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Besides having the appropriate amino acid sequence to perform the function of proteins, it is important to have correct conformation after this sequence to process. To consist of this conformation depends on the amino acid sequence at the primary structure, hydrophobic interaction, chaperones and enzymes in charge of folding etc. Misfolded proteins are not functional and tend to be aggregated. Cysteine originating disulfide cross-links make stable this conformation of functional proteins. When two of the cysteine amino acids come side by side, disulfide bond is established that forms a cystine bridge. Due to this feature cysteine plays an important role on the formation of three-dimensional structure of many proteins. There are two cysteine amino acids (C44, C69) in the Tol-A-III protein. Unlike protein disulfide bonds from within his own, any non-specific cystine bridge causes a change in the three dimensional structure of the protein. Proteins can be expressed in various host cells as directly or fusion (chimeric). As a result of overproduction of the recombinant proteins, aggregation of insoluble proteins in the host cell can occur by forming a crystal structure called inclusion body. In general fusion proteins are produced for provide affinity tags to make proteins more soluble and production of some toxic proteins via fusion protein expression system like pTolT. Proteins can be modified by using a site-directed mutagenesis. By this way, creation of non-specific disulfide crosslinks can be prevented at fusion protein expression system via the present cysteine replaced by another amino acid such as serine, glycine or etc. To do this, we need; a DNA molecule that contains the gene that encodes for the target protein, required primers for mutation to be designed according to site directed mutagenesis reaction. This study was aimed to be replaced cysteine encoding codon TGT with serine encoding codon AGT. For this sense and reverse primers designed (given below) and used site-directed mutagenesis reaction. Several new copy of the template plasmid DNA has been formed with above mentioned mutagenic primers via polymerase chain reaction (PCR). PCR product consists of both the master template DNA (wild type) and the new DNA sequences containing mutations. Dpn-l endonuclease restriction enzyme which is specific for methylated DNA and cuts them to the elimination of the master template DNA. E. coli cells obtained after transformation were incubated LB medium with antibiotic. After purification of plasmid DNA from E. coli, the presence of the mutation was determined by DNA sequence analysis. Developed this new plasmid is called PtolT-δ.

Keywords: site directed mutagenesis, Escherichia coli, pTolT, protein expression

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Authors: Siddhant Sethi, Yasuharu Takashima, Shigetaka Nakamura, Kenzo Fujimoto

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Site-directed mutagenesis is a renowned technique to introduce specific mutations in the genome. To achieve site-directed mutagenesis, many chemical and enzymatic approaches have been reported in the past like disulphite induced genome editing, CRISPR-Cas9, TALEN etc. The chemical methods are invasive whereas the enzymatic approaches are time-consuming and expensive. Most of these techniques are unusable in the cellular application due to their toxicity and other limitations. Photo-chemical cytosine deamination, introduced in 2010, is one of the major technique for enzyme-free single-point mutation of cytosine to uracil in DNA and RNA, wherein, 3-cyanovinylcarbazole nucleoside (CNVK) containing oligodeoxyribonucleotide (ODN) having CNVK at -1 position to that of target cytosine is reversibly crosslinked to target DNA strand using 366 nm and then incubated at 90ºC to accommodate deamination. This technique is superior to enzymatic methods of site-directed mutagenesis but has a disadvantage that it requires the use of high temperature for the deamination step which restricts its applicability in the in vivo applications. This study has been focused on improving the technique by reducing the temperature required for deamination. Firstly, the photo-cross-linker, CNVK has been modified by replacing cyano group attached to vinyl group with methyl ester (OMeVK), amide (NH2VK), and carboxylic acid (OHVK) to observe the acceleration in the deamination of target cytosine cross-linked to vinylcarbazole derivative. Among the derivatives, OHVK has shown 2 times acceleration in deamination reaction as compared to CNVK, while the other two derivatives have shown deceleration towards deamination reaction. The trend of rate of deamination reaction follows the same order as that of hydrophilicity of the vinylcarbazole derivatives. OHVK being most hydrophilic has shown highest acceleration while OMeVK is least hydrophilic has proven to be least active for deamination. Secondly, in the related study, the counter-base of the target cytosine, guanine has been replaced by inosine, 2-aminopurine, nebularine, and 5-nitroindole having distinct hydrogen bonding patterns with target cytosine. Among the ODNs with these counter bases, ODN with inosine has shown 12 fold acceleration towards deamination of cytosine cross-linked to CNVK at physiological conditions as compared to guanosine. Whereas, when 2-aminopurine, nebularine, and 5-nitroindole were used, no deamination reaction took place. It can be concluded that inosine has potential to be used as the counter base of target cytosine for the CNVK mediated photo-cross-linking induced deamination of cytosine. The increase in rate of deamination reaction has been attributed to pattern and number of hydrogen bonding between the cytosine and counter base. One of the important factor is presence of hydrogen bond between exo-cyclic amino group of cytosine and the counter base. These results will be useful for development of more efficient technique for site-directed mutagenesis for C → U transformations in the DNA/RNA which might be used in the living system for treatment of various genetic disorders and genome engineering for making designer and non-native proteins.

Keywords: C to U transformation, DNA editing, genome engineering, ultra-fast photo-cross-linking

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Authors: Han-Bin Kim, Sooim Shin, Moonsung Choi

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There is a growing interest in introducing a desired functional group on enzymes in the field of protein engineering. In here, various redox centers were newly created using histidine tag, which is widely used for protein purification, plus cobalt in one of cupredoxins, amicyanin. The coordination of Cobalt-His tag and reactivity of the Co²⁺ loaded His-tag also were characterized. 3xHis-tag, 6xHis-tag, and 9xHis-tag were introduced on amicyanin by site-directed mutagenesis, and then Co²⁺ was loaded on each His-tagged amicyanin. The spectral changes at 330 nm corresponding to cobalt binding on His-tag site indicated the binding ratio of 3xHis-tag, 6xHis-tag, and 9xHis-tag to cobalt as 1:1, 1:2, 1:3 respectively. Based on kinetic studies of binding cobalt to 3xHis-tag, 6xHis-tag, and 9xHis-tagged amicyanin, the nature of the sites was elucidated. In addition, internal electron transfer properties between Cu¹⁺ site and engineered site of amicyanin were determined. These results provide insight into improvement of metal coordination and alternation of the redox properties of metal as a new catalytic site on proteins.

Keywords: amicyanin, cobalt, histidine, protein engineering

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Authors: D. Shehu, Z. Alias

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Glutathione S-transferases (GSTs) are family of enzymes that function in the detoxification of variety of electrophilic substrates. In the present work, we report a novel zeta-like GST (designated as KKSG9) from the biphenyl/polychlorobiphenyl degrading organism Acidovorax sp. KKS102. KKSG9 possessed low sequence similarity but similar biochemical properties to zeta class GSTs. The gene for KKSG9 was cloned, purified and biochemically characterized. Functional analysis showed that the enzyme exhibits wider substrate specificity compared to most zeta class GSTs by reacting with 1-chloro-2,4-dinitrobenzene (CDNB), p-nitrobenzyl chloride (NBC), ethacrynic acid (EA), hydrogen peroxide, and cumene hydroperoxide (CuOOH). The enzyme also displayed dehalogenation function against dichloroacetate (a common substrate for zeta class GSTs) in addition to permethrin, and dieldrin. The functional role of Tyr12 was also investigated by site-directed mutagenesis. The mutant (Y12C) displayed low catalytic activity and dehalogenation function against all the substrates when compared with the wild type. Kinetic analysis using NBC and GSH as substrates showed that the mutant (Y12C) displayed a higher affinity for NBC when compared with the wild type, however, no significant change in GSH affinity was observed. These findings suggest that the presence of tyrosine residue in the motif might represent an evolutionary trend toward improving the catalytic activity of the enzyme. The enzyme as well could be useful in the bioremediation of various types of organochlorine pollutants.

Keywords: Acidovorax sp. KKS102, bioremediation, glutathione s-transferase, site-directed mutagenesis, zeta

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Authors: Iman Farasat, Howard M. Salis

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The CRISPR/Cas9 system has revolutionized genome engineering by enabling site-directed and high-throughput genome editing, genome insertion, and gene knockdowns in several species, including bacteria, yeast, flies, worms, and human cell lines. This technology has the potential to enable human gene therapy to treat genetic diseases and cancer at the molecular level; however, the current CRISPR/Cas9 system suffers from seemingly sporadic off-target genome mutagenesis that prevents its use in gene therapy. A comprehensive mechanistic model that explains how the CRISPR/Cas9 functions would enable the rational design of the guide-RNAs responsible for target site selection while minimizing unexpected genome mutagenesis. Here, we present the first quantitative model of the CRISPR/Cas9 genome mutagenesis system that predicts how guide-RNA sequences (crRNAs) control target site selection and cleavage activity. We used statistical thermodynamics and law of mass action to develop a five-step biophysical model of cas9 cleavage, and examined it in vivo and in vitro. To predict a crRNA's binding specificities and cleavage rates, we then compiled a nearest neighbor (NN) energy model that accounts for all possible base pairings and mismatches between the crRNA and the possible genomic DNA sites. These calculations correctly predicted crRNA specificity across 5518 sites. Our analysis reveals that cas9 activity and specificity are anti-correlated, and, the trade-off between them is the determining factor in performing an RNA-mediated cleavage with minimal off-targets. To find an optimal solution, we first created a scheme of safe-design criteria for Cas9 target selection by systematic analysis of available high throughput measurements. We then used our biophysical model to determine the optimal Cas9 expression levels and timing that maximizes on-target cleavage and minimizes off-target activity. We successfully applied this approach in bacterial and mammalian cell lines to reduce off-target activity to near background mutagenesis level while maintaining high on-target cleavage rate.

Keywords: biophysical model, CRISPR, Cas9, genome editing

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Authors: Syeda Kiran Shahzadi, Nasir Mahmood, Muhammad Abdul Qadir

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In the current research, three recombinant human interferon alpha-2b proteins (two modified and one normal form) were produced and Pegylated with an aim to produce more effective drugs against viral infections and cancers. The modified recombinant human interferon alpha-2b proteins were produced by site-directed modifications of interferon alpha 2b gene, targeting the amino acids at positions ‘R23’ and ‘H34’. The resulting chemically modified and unmodified forms of human interferon alpha 2b were conjugated with methoxy-polyethylene glycol propanealdehyde (400 KDa) and methoxy-polyethylene glycol succinimidyl succinate (400 KDa). Pegylation of normal and modified forms of Interferon alpha-2b prolong their release time and enhance their efficacy. The conjugation of PEG with modified and unmodified human interferon alpha 2b protein drugs was also characterized with 1H-NMR, HPLC, and SDS-PAGE. Antiproliferative assays of modified and unmodified forms of drugs were performed in cell based bioassays using MDBK cell lines. The results indicated that experimentally produced recombinant human interferon alpha-2b proteins were biologically active and resulted in significant inhibition of cell growth.

Keywords: protein refolding, antiproliferative activities, biomedical applications, human interferon alpha-2b, pegylation, mPEG-propionaldehyde, site directed mutagenesis, E. coli expression

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Authors: Sanaa Bardaweel

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Inorganic polyphosphate (poly P) is present in all living forms tested to date, from each of the three kingdoms of life. Studied mainly in prokaryotes, poly P and its associated enzymes are vital in diverse basic metabolism, in at least some structural functions and, notably, in stress responses. These plentiful and unrelated roles for poly P are probably the consequence of its presence in life-forms early in evolution. The genomes of many bacterial species, including pathogens, encode a homologue of a major poly P synthetic enzyme, poly P kinase 1 (PPK1). Genetic deletion of ppk1 results in reduced poly P levels and loss of pathogens virulence towards protozoa and animals. Thus far, no PPK1 homologue has been identified in higher-order eukaryotes and, therefore, PPK1 represents a novel target for chemotherapy. The idea of the current study is to purify the PPK1 from Escherichia coli to homogeneity in order to study the effect of active site point mutations on PPK1 catalysis via the application of site-directed mutagenesis strategy. The knowledge obtained about the active site of PPK1 will be utilized to characterize the catalytic and kinetic mechanism of PPK1 with model substrates. Comprehensive understanding of the enzyme kinetic mechanism and catalysis will be used to design and screen a library of synthetic compounds for potential discovery of selective PPK1-inhibitors.

Keywords: antimicobial, Escherichia coli, inorganic polyphosphate, PPK1-inhibitors

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Authors: Mark-Adam Kellerman

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Proteins are capable of exploring vast mutational spaces. This makes it difficult for protein engineers to devise rational methods to improve stability and function via mutagenesis. Deciding which residues to mutate requires knowledge of the characteristics they elicit. We probed the characteristics of residues in granulocyte-colony stimulating factor (G-CSF) using a thermal melt (from 295K to 323K) to denature it in a 700 MHz Bruker spectrometer. These characteristics included dynamics, micro-environmental changes experienced/ induced during denaturing and structure-function relationships. 15N-1H HSQC experiments were performed at 2K increments along with this thermal melt. We observed that dynamic residues that also undergo a lot of change in their microenvironment were predominantly in unstructured regions. Moreover, we were able to identify four residues (G4, A6, T133 and Q134) that we class as high priority targets for mutagenesis, given that they all appear in both the top 10% of measures for environmental changes and dynamics (∑Δ and ∆PI). We were also able to probe these NMR observables and combine them with molecular dynamics (MD) to elucidate what appears to be an opening motion of G-CSFs binding site III. V48 appears to be pivotal to this opening motion, which also seemingly distorts the loop region between helices A and B. This observation is in agreement with previous findings that the conformation of this loop region becomes altered in an aggregation-prone state of G-CSF. Hence, we present here an approach to profile the characteristics of residues in order to highlight their potential as rational mutagenesis targets and their roles in important conformational changes. These findings present not only an opportunity to effectively make biobetters, but also open up the possibility to further understand epistasis and machine learn residue behaviours.

Keywords: protein engineering, rational mutagenesis, NMR, molecular dynamics

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Authors: Silmy A. Humaira'

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There is an issue about Asian students including Indonesian students that tend to behave passively in the classroom and depend on the teachers’ instruction. Regarding this statement, this study attempts to address the Indonesian high school students’ attitudes on whether they have initiative and be responsible for their learning out of the classroom and if so, why. Therefore, 30 high school students were asked to fill out the questionnaires and interviewed in order to figure out their attitudes towards self-directed learning. The descriptive qualitative research analysis adapted Knowles’s theory (1975) about Self-directed learning (SDL) to analyze the data. The findings show that the students have a potential to possess self-directed learning through ICT, but they have difficulties in choosing appropriate learning strategy, doing self-assessment and conducting self-reflection. Therefore, this study supports the teacher to promote self-directed learning instruction for successful learning by assisting students in dealing with those aforementioned problems. Furthermore, it is expected to be a beneficial reference which gives new insights on the self-directed learning practice in specific context.

Keywords: ICT, learning autonomy, students’ attitudes, self-directed learning

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Authors: Timna Mayer

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This paper explores the vast potential of self-directed learning in violin pedagogy. Based in practice and drawing on concepts from neuropsychology, the author, a violinist and teacher, outlines five learning principles. Self-directed learning is defined as an ongoing process based on problem detection, definition, and resolution. The traditional roles of teacher and student are reimagined within this context. A step-by-step guide to applied self-directed learning suggests a model for both teachers and students that realizes student independence in the classroom, leading to higher-level understanding and more robust performance. While the value of self-directed learning is well-known in general pedagogy, this paper is novel in applying the approach to the study of musical performance, a field which is currently dominated by habit and folklore, rather than informed by science.

Keywords: neuropsychology and musical performance, self-directed learning, strategic problem solving, violin pedagogy

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Authors: G. Ultanbekova, Zh. Suleimenova, Zh. Rakhmetova, G. Mombekova, S. Mantieva

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Plant Growth Promoting Rhizobacteria (PGPR) are a group of free-living bacteria that colonize the rhizosphere, enhance plant growth of many cereals and other important agricultural crops and protect plants from disease and abiotic stresses through a wide variety of mechanisms. The use of PGPR has been proven to be an environmentally sound way of increasing crop yields by facilitating plant growth. In the present study, strain improvement of PGPR isolates were carried out by chemical mutagenesis for the improvement of growth and yield of lima bean. Induced mutagenesis is widely used for the selection of microorganisms producing biologically active substances and further improving their activities. Strain improvement is usually done by classical mutagenesis which involves exposing the microbes to chemical or physical mutagens. The strains of Pseudomonas putida 4/1, Azotobacter chroococcum Р-29 and Bacillus subtilis were subjected to mutation process for strain improvement by treatment with a chemical agent (sodium nitrite) to cause mutation and were observed for its consequent action on the seeds germination and plant growth of lima bean (Phaseolus lunatus). Bacterial mutant strains of Pseudomonas putida M-1, Azotobacter chroococcum M-1 and Bacillus subtilis M-1, treated with sodium nitrite in the concentration of 5 mg/ml for 120 min, were found effective to enhance the germination of lima bean seeds compared to parent strains. Moreover, treatment of the lima bean seeds with a mutant strain of Bacillus subtilis M-1 had a significant stimulation effect on plant growth. The length of the stems and roots of lima bean treated with Bacillus subtilis M-1 increased significantly in comparison with parent strain in 1.6 and 1.3 times, respectively.

Keywords: chemical mutagenesis, germination, kidney bean, plant growth promoting rhizobacteria (PGPR)

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Authors: Rosaline Govender

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This research study is an exploration of the self-directed professional development of teachers who teach in public schools in an era of democracy and educational change in South Africa. Amidst an ever-changing educational system, the teachers in this study position themselves as self-directed teacher-learners where they adopt particular learning practices which enable change within the broader discourses of public schooling. Life-story interviews were used to enter into the private and public spaces of five teachers which offer glimpses of how particular systems shaped their identities, and how the meanings of self-directed teacher-learner shaped their learning practices. Through the Multidimensional framework of analysis and interpretation the teachers’ stories were analysed through three lenses: restorying the field texts - the self through story; the teacher-learner in relation to social contexts, and practices of self-directed learning.This study shows that as teacher-learners learn for change through self-directed learning practices, they develop their agency as transformative intellectuals, which is necessary for the reworking of South African public schools.

Keywords: professional development, professionality, professionalism, self-directed learning

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Authors: Hunmin Jung, Michael Hawkins, Seongmin Lee

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The exocyclic amines of nucleobases can undergo deamination by various DNA damaging agents such as reactive oxygen species, nitric oxide, and water. The deamination of guanine and adenine generates the promutagenic xanthine and hypoxanthine, respectively. The exocyclic amines of bases in DNA are hydrogen bond donors, while the carbonyl moiety generated by the base deamination acts as hydrogen bond acceptors, which can alter base pairing properties of the purines. Xanthine is known to base pair with both cytosine and thymine, while hypoxanthine predominantly pairs with cytosine to promote A to G mutations. Despite the known promutagenicity of the major deaminated purines, structures of DNA polymerase bypassing these lesions have not been reported. To gain insights into the deaminated-induced mutagenesis, we solved crystal structures of human DNA polymerase η (polη) catalyzing across xanthine and hypoxanthine. In the catalytic site of polη, the deaminated guanine (i.e., xanthine) forms three Watson-Crick-like hydrogen bonds with an incoming dCTP, indicating the O2-enol tautomer of xanthine involves in the base pairing. The formation of the enol tautomer appears to be promoted by the minor groove contact by Gln38 of polη. When hypoxanthine is at the templating position, the deaminated adenine uses its O6-keto tautomer to form two Watson-Crick hydrogen bonds with an incoming dCTP, providing the structural basis for the high promutagenicity of hypoxanthine.

Keywords: DNA damage, DNA polymerase, deamination, mutagenesis, tautomerization, translesion synthesis

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Authors: Yu-Tzu Chen, Luh-Maan Chang

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Construction is an inherently risky industry. The projects have been dominated by reactive actions owing to non-routine in nature. The on-site activities are especially crucial for successful project control. In order to alter actions from reactive to proactive, this paper presents an on-site data collection system utilizing advanced technology RFID and GPS in assisting on-site management with near real time progress monitoring.

Keywords: On-Site management, progress monitoring, RFID, GPS

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Authors: Jocelyn D. Perreault

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An exploratory study has been launched in 2013-2014 in the province of Quebec, the state of Vermont (USA) and the region of Zaer in Morocco. We have realized three first case studies in order to better understand the marketing strategies of starting up vineries, which are defined as having a maximum of five years of operations. The methodology used consisted of visiting the vineyards; conducting semi-directed interviews with owner-managers; visiting points-of-sale of the wines and analysing the web sites using an assessment grid. The results indicate many differences between the three firms in their use of their web sites. More precisely, we have noticed that: -The Quebec vineyard uses its web site in collaboration with the touristic actors of its region and the association of the wine makers of the province of Quebec.Positioning is as a touristic attraction. -In comparison,the Moroccan firm limits the content of the web site to itself and its activities and somehow to the wine industry.Positioning is as a wine specialist. -The american firm associated its web site more to farm markets actors and activities of the region.Positioning is as an agricultural actor. -The positionings of the three vineyards are very different from each others and will be discussed more thoroughly during the presentation to better understand the use of web sites, thus contributing to the «brand image». -Improvements to the three web sites have been identified and suggested by more than a hundred of persons using the same grid and comprising students of bachelor and MBA degrees from our university. In general, the web sites have been considered satisfying but requiring several improvements at different levels. Changes or updates have been observed for the Quebec winery web site but practically no changes have been made to the others in the last months. The assessment grid will be presented in more details as well as the global and the partial scores given by the respondents. In conclusion, we have noticed that only one winery is considered as a «heavy and strategic user» of its web site and of Facebook and Twitter.

Keywords: web site, wineries, marketing, positioning, starting up strategies

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Authors: Mekwa Eme, Anya chukwuma

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The act of evaluating a particular site physically and socially in order to create a good design solution that will address the physical and interior environment of the location is known as architectural site analysis. This essay will describe site analysis as a useful design component. According to the introduction and supporting research, site evaluation and analysis are crucial to good design in terms of topography, orientation, site size, accessibility, rainfall, wind direction, and times of sunrise and sunset. Methodology: Both quantitative and qualitative analyses are used in this paper. The primary and secondary types of data collection are as follows. This information was gathered via the case study approach, already published literature, journals, the internet, a local poll, oral interviews, inquiries, and in-person interviews. The purpose of this is to clarify the benefits of site analysis for the design process and its implications for the working or building stage. Results: Each site's criteria are unique in terms of things like soil, plants, trees, accessibility, topography, and security. This will make it easier for the architect and environmentalist to decide on the idea, shape, and supporting structures of the design. It is crucial because before any design work is done, the nature of the target location will be determined through site visits and research. The location, contours, site features, and accessibility are just a few of the topics included in this site study. In order for students and working architects to understand the nature of the site they will be working on, site analysis is a key component of architectural education. The building's orientation, the site's circulation, and the sustainability of the site may all be determined with thorough research of the site's features.

Keywords: analysis, climate, statistics, design

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Authors: Heri Kuswanto, Philipp Sibbertsen, Irhamah

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A Wald type test to distinguish between long memory and ESTAR nonlinearity has been developed. The test uses a directed-Wald statistic to overcome the problem of restricted parameters under the alternative. The test is derived from a model specification i.e. allows the transition parameter to appear as a nuisance parameter in the transition function. A simulation study has been conducted and it indicates that the approach leads a test with good size and power properties to distinguish between stationary long memory and ESTAR.

Keywords: directed-Wald test, ESTAR, long memory, distinguish

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Authors: Faisal Alazzaz, Andrew Whyte

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Off-site construction methods have played an important role in the construction sector in the past few decades. It is increasingly becoming a major alternative technique and strategic direction compared to traditional in-situ method. It produces a significant amount of value for the construction industry and the economy more generally. To date, an impressive number of studies have been lunched on the perceived perception of off-site construction. However, it seems that a quantifying benefit on the offsite construction area is lacking. Therefore, this paper examines the recent research literature on the benefits of off- site construction and provides future direction. In the beginning, this paper provides a brief history and current value of the off-site construction followed by a detailed discussion on the benefit of off-site construction. These benefits include but not limited to time saving, quality improvement, relieving skills shortages, cost reduction and productivity improvement. Toward this end, off-site construction should learn from other productive industry similar to services or manufacturing industry by applying operational management tools and techniques with extensive focus on employee empowerment will shed the light on future uptake of Off-site construction. This study is of value in providing scholars have a clear picture of perceived benefit of off-site construction research and give an opportunities for future uptake of off-site method.

Keywords: building projects, employer empowerment, off-site construction benefits, productivity

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Authors: Yassir AbdelRazig

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Evolutionary optimization methods such as genetic algorithms have been used extensively for the construction site layout problem. More recently, ant colony optimization algorithms, which are evolutionary methods based on the foraging behavior of ants, have been successfully applied to benchmark combinatorial optimization problems. This paper proposes a formulation of the site layout problem in terms of a sequencing problem that is suitable for solution using an ant colony optimization algorithm. In the construction industry, site layout is a very important planning problem. The objective of site layout is to position temporary facilities both geographically and at the correct time such that the construction work can be performed satisfactorily with minimal costs and improved safety and working environment. During the last decade, evolutionary methods such as genetic algorithms have been used extensively for the construction site layout problem. This paper proposes an ant colony optimization model for construction site layout. A simple case study for a highway project is utilized to illustrate the application of the model.

Keywords: ant colony, construction site layout, optimization, genetic algorithms

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Authors: Yaxiao Zhang, Yangzhou Chen

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This study investigate the formation problem for high-order continuous-time multi-robot with bounded symmetric time-varying delay protocol under switched directed communication topology. By using a linear transformation, the formation problem is transformed to stability analysis of a switched delay system. Under the assumption that each communication topology has a directed spanning tree, sufficient conditions are presented in terms of linear matrix inequalities (LMIs) that the multi-robot system can achieve a desired formation by the trade-off among the pre-exist topologies with the help of the scheme of average dwell time. A numeral example is presented to illustrate the effectiveness of the obtained results.

Keywords: multi-robot systems, formation, switched directed topology, symmetric time-varying delay, average dwell time, linear matrix inequalities (lmis)

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Authors: Ruichong Zhang

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

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

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Authors: Son Nguyen, Thanh Van, Ly Le

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Ca2+ - dependent calmodulin kinase IIa (CaMKIIa) has recently been found to associate with protein tau missorting and polymerization in Alzheimer’s Disease (AD). However, there has yet inhibitors targeting CaMKIIa to investigate the correlation between CaMKIIa activity and protein tau polymer formation. Combining virtual screening and our statistics in binding contribution scoring function (BCSF), we rationally identified potential compounds that bind to specific CaMKIIa active site and specificity-affinity distribution of the ligand within the active site. Using molecular dynamics simulation, we identified structural stability of CaMKIIa and potent inhibitors, and site-directed bonding, separating non-specific and specific molecular interaction features. Despite of variation in confirmation of simulation time, interactions of the potent inhibitors were found to be strongly associated with the unique chemical features extracted from molecular binding poses. In addition, competitive inhibitors within CaMKIIa showed an important molecular recognition pattern toward specific ligand features. Our approach combining virtual screening with BCSF may provide an universally applicable method for precise identification in the discovery of compounds.

Keywords: Alzheimer’s disease, Ca 2+ -dependent calmodulin kinase IIa, protein tau, molecular docking

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Authors: Rameez Ahmad, Aqib Mehmood, Imran Khan

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Self-directed car (SDC) that can drive itself from one fact to another without support from a driver. Certain trust that self-directed car obligate the probable to transform the transportation manufacturing while essentially removing coincidences, and cleaning up the environment. This study realizes the effects that SDC (also called a self-driving, driver or robotic) vehicle travel demands and ride scheme is likely to have. Without the typical obstacles that allows detection of a audio vision based hardware and software construction (It (SDC) and cost benefits, the vehicle technologies, Gold (Generic Obstacle and Lane Detection) to a knowledge-based system to predict their potential and consider the shape, color, or balance) and an organized environment with colored lane patterns, lane position ban. Discovery the problematic consequence of (SDC) on GT (grand trunk road) road and brand the car further effectual.

Keywords: SDC, gold, GT, knowledge-based system

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Authors: Zs. Csanádi, A. Szabó Nagy, J. Szabó, J. Erdős

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The main objective of this study was to assess the annual concentration and seasonal variation of benzo(a)pyrene (BaP) associated with PM10 in an urban site of Győr and in a rural site of Sarród in the sampling period of 2008–2012. A total of 280 PM10 aerosol samples were collected in each sampling site and analyzed for BaP by gas chromatography method. The BaP concentrations ranged from undetected to 8 ng/m3 with the mean value of 1.01 ng/m3 in the sampling site of Győr, and from undetected to 4.07 ng/m3 with the mean value of 0.52 ng/m3 in the sampling site of Sarród, respectively. Relatively higher concentrations of BaP were detected in samples collected in both sampling sites in the heating seasons compared with non-heating periods. The annual mean BaP concentrations were comparable with the published data of different other Hungarian sites.

Keywords: air quality, benzo(a)pyrene, PAHs, polycyclic aromatic hydrocarbons

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Authors: Sinem Bulkan

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This study introduces the construct of psychopathic manager behaviour and aims for the development of psychopathic manager behaviour (Psycho-Man B) measure. The study also aims for the understanding of the relationship between psychopathic manager behaviour and workplace deviance while investigating the mediating role of a revenge motive and the moderating roles of the core self-evaluations and the attitude importance. Data were collected from 519 employees from a wide variety of jobs and industries who currently work for or previously worked for a manager in a collectivist culture, Turkey. Psycho-Man B Measure was developed resulting in five dimensions as opposed to the proposed ten dimensions. Simple linear and hierarchical regression analyses were conducted to test the hypotheses. The results of simple linear regression analyses showed that psychopathic manager behaviour was positively significantly related to supervisor-directed and organisation-directed deviance. Revenge motive towards the manager partially mediated the relationship between psychopathic manager behaviour and supervisor-directed deviance. Similarly, revenge motive towards the organisation partially mediated the relationship between psychopathic manager behaviour and organisation-directed deviance. Furthermore, no support was found for the expected moderating role of core self-evaluations in the revenge motive towards the manager-supervisor-directed deviance and revenge motive towards the organisation-organisation-directed deviance relationships. Attitude importance moderated the relationship between revenge motive towards the manager and supervisor-directed deviance; revenge motive towards the organisation and organisation-directed deviance. Moderated-mediation hypotheses were not supported for core self-evaluations but were supported for the attitude importance. Additional analyses for sub-dimensions were conducted to further examine the hypotheses. Demographic variables were examined through independent samples t-tests, and one way ANOVA. Finally, findings are discussed; limitations, suggestions and implications are presented. The major contribution of this study is that ‘psychopathic manager behaviour’ construct was introduced to the literature and a scale for the reliable identification of psychopathic manager behaviour was developed in to evaluate managers’ level of sub-clinical psychopathy in the workforce. The study introduced that employees engage in different forms of supervisor-directed deviance and organisation-directed deviance depending on the level of the emotions and personal goals. Supervisor-directed deviant behaviours and organisation-directed deviant behaviours became distinct in a way as impulsive and premeditated, active or passive, direct and indirect actions. Accordingly, it is important for organisations to notice that employees’ level of affective state and attitude importance for psychopathic manager behaviours predetermine the certain type of employee deviant behaviours.

Keywords: attitude importance, core self evaluations, psychopathic manager behaviour, revenge motive, workplace deviance

Procedia PDF Downloads 232

Authors: Wah Wah Min

Abstract:

The relative abundance and diversities of gastropods and crabs were assessed for mangrove areas of Ko Libong, Kantang district, Trang, Thailand in June 2022. Two sample sites (I and II) were studied. The site I was replanted under ecotourism, whereas site II represented the protected natural restored mangroves. This study is aimed to assess faunal diversity and how it could become re-established and resemble to natural restored mangroves. There was one sample plot at each study site with the dimension (10m x 25m) in study site I and (20m x 30m) in site II. The sample was randomly taken from each plot by using a quadrate measuring at (1 m2) in site I and (3m2) in site II; there were four quadrates in total of each site. The species richness (S), Shannon Index (H’) and Evenness Index (J’), vegetative measurements and physico-chemical parameters were calculated for each site. Seventeen gastropod species belonged to 11 families and six crab species under two families, which were collected in both study sites. Overall, in gastropod species, the highest relative abundance of Nerita planospira exhibited (53.45%, category C) with lower population density (1.61 individuals/m2), whichwas observed in study site II and for crab species, Parasesarma plicatum (83.33%, category C) with lower population density (0.33 individuals/m2). The diversity indices of gastropod species at the study site I was calculated higher indicating by (S= 12, H’= 2.27, J’ and SDI=0.91) compared to study site II (S= 7, H’= 1.22, J’ and SDI=0.63, 0.62). For the crabs, (S= 4, H’=1.33, J’ and SDI=0.96, 0.9) in study site I and (S= 2, H’=0.64, J’ and SDI=0.92, 0.67) in site II. Overall, the higher species diversity indices of study site I can be categorized “very equally” with a very good category according to evenness criteria (>0.81). This can be gained by increasing restoration sites through an ecotourism replanting program for achieving the goals of sustainable development for mangrove conservation and long-term studies are required to confirm this hypothesis.

Keywords: biodiversity, ecotourism, restoration, population

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Authors: Babatunde Femi Bakare

Abstract:

Groundwater quality was evaluated at a site for three years after the site was used for entrenchment of Ventilated Improved Pit (VIP) latrine sludge. Analysis performed on the soil characteristics at the entrenchment site indicated that, the soils at the entrenchment site are predominantly sandy. Depth of the water table at the entrenchment site was found to be approximately five meters. Five monitoring boreholes were dug along the perimeter of the sludge trenches and water samples taken from these monitoring boreholes were analyzed for pH, conductivity, sodium ions, chloride ions, phosphate, nitrate, ammonia, and bacteriological analysis. The results obtained from the analysis conducted were compared with the South African Bureau of Standards for drinking water and it was found that the parameters analyzed falls below the specified range. The data obtained from this study indicate that, given the relatively high sludge loading rates, poor soil quality, and the duration of the groundwater quality monitoring, it is unlikely that contamination of groundwater at the entrenchment site will be a major concern. However, caution is advised in extrapolating these results to other locations.

Keywords: boreholes, contamination, entrenchment, groundwater quality, VIP latrines

Procedia PDF Downloads 374

Authors: F. Karim

Abstract:

In this paper, triangular lattice index-guiding photonic crystal fibers (PCFs) are synthesized to compensate the chromatic dispersion of a single mode fiber (SMF-28) for an 80 km optical link operating at 1.55 µm, by using the directed tabu search algorithm. Hole-to-hole distance, circular air-hole diameter, solid-core diameter, ring number and PCF length parameters are optimized for this purpose. Three Synthesized PCFs with different physical parameters are compared in terms of their objective functions values, residual dispersions and compensation ratios.

Keywords: triangular lattice index-guiding photonic crystal fiber, dispersion compensation, directed tabu search, synthesis

Procedia PDF Downloads 402

Authors: Osama Amer Mohyeldin Mohamed

Abstract:

Qubbet el-Hawa North (QHN) is an example of an archaeological site in West-Aswan and is not opened for the public yet and has been under excavation since its discovery in 2013 as a result of the illegal digging that happened in many sites in Egypt because of the unstable situation and the absence of security. The site has the potential to be one of the most attractive sites in Aswan. Moreover, it deserves to be introduced to the visitors in a good manner appropriate to its great significance. Both interpretation and presentation are crucial inseparable tools that are communicating the archaeological site significance to the public and raising their awareness. Moreover, it helps them to understand the past and appreciate the archaeological assets. People will never learn or see anything from ancient remains unless it was explained. They would only look to it as ancient and charming. They expect a story, and more than knowledge, authenticity, or even supporting preservation actions, they want to enjoy and be entertained. On the other hand, a lot of archaeologists believe that planning an archaeological site for entertaining the visitors deteriorates it and effects its authenticity. Thus, it represents a challenge to design a model for visitors’ experience that meets their expectations and needs with safeguarding the site’s integrity. The article is presenting a proposal for interpretation plan to the site of Qubbet el-Hawa North.

Keywords: heritage interpretation and presentation, archaeological site management, qubbet el-hawa North, local community engagement, accessibility

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Authors: Mehri Bagherkazemi, Naser Hafezi Moghaddas

Abstract:

This study investigates the environmental impact of the disposal site located in Shahrood city, focusing on the geological characteristics of the region. Shahrood's disposal site primarily consists of limestone bedrock, overlaid by substantial alluvial deposits. The area's highly permeable soil is anticipated to have a significant influence on groundwater pollution. Spanning 52 hectares, the Shahrood disposal site is situated in the eastern sector of the city. Historically, waste disposal took place on the surface, but recent practices involve on-site trenching. This research involved the collection of soil and water samples near the disposal site, with subsequent analysis of 11 soil samples and 3 water samples. The soil's particle size distribution was determined, and comprehensive analyses were conducted for 35 elements in the soil and 42 elements in water. The study combines the results of these tests with field investigations to evaluate the landfill's impact on the surrounding soil and groundwater contamination.

Keywords: environmental geology, environmental impact assessment, disposal site, heavy metals contamination

Procedia PDF Downloads 36