Search results for: multiplex polymerase chain reaction

Authors: O. Ubani, H. I. Atagana, M. S. Thantsha, R. Adeleke

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The oil sludge components (polycyclic aromatic hydrocarbons, PAHs) have been found to be cytotoxic, mutagenic and potentially carcinogenic and microorganisms such as bacteria and fungi can degrade the oil sludge to less toxic compounds such as carbon dioxide, water and salts. In the present study, we isolated different bacteria with PAH-degrading potentials from the co-composting of oil sludge and different animal manure. These bacteria were isolated on the mineral base medium and mineral salt agar plates as a growth control. A total of 31 morphologically distinct isolates were carefully selected from 5 different compost treatments for identification using polymerase chain reaction (PCR) of the 16S rDNA gene with specific primers (16S-P1 PCR and 16S-P2 PCR). The amplicons were sequenced and sequences were compared with the known nucleotides from the gene bank database. The phylogenetical analyses of the isolates showed that they belong to 3 different clades namely Firmicutes, Proteobacteria and Actinobacteria. These bacteria identified were closely related to genera Bacillus, Arthrobacter, Staphylococcus, Brevibacterium, Variovorax, Paenibacillus, Ralstonia and Geobacillus species. The results showed that Bacillus species were more dominant in all treated compost piles. Based on their characteristics these bacterial isolates have high potential to utilise PAHs of different molecular weights as carbon and energy sources. These identified bacteria are of special significance in their capacity to emulsify the PAHs and their ability to utilize them. Thus, they could be potentially useful for bioremediation of oil sludge and composting processes.

Keywords: bioaugmentation, biodegradation, bioremediation, composting, oil sludge, PAHs, animal manures

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Authors: Sang-Woo Han, Jong-Shik Shin

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w-Transaminase (w-TA) is broadly used for synthesizing chiral amines with a high enantiopurity. However, the reaction mechanism of w-TA has been not well studied, contrary to a-transaminase (a-TA) such as AspTA. Here, we propose in silico model on the reaction mechanism of w-TA. Based on the modeling results which showed large free energy gaps between external aldimine and quinonoid on deamination (or ketimine and quinonoid on amination), withdrawal of Ca-H seemed as a critical step which determines the reaction rate on both amination and deamination reactions, which is consistent with previous researches. Hyperconjugation was also observed in both external aldimine and ketimine which weakens Ca-H bond to elevate Ca-H abstraction.

Keywords: computational modeling, reaction intermediates, w-transaminase, in silico model

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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: Saqib Ali, Man-Qun Wang

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The Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae), is a major pest of pines and it is also the key vector of the exotic pinewood nematode in China. In the present study, we cloned, expressed, and purified a chemosensory protein (CSP) in M. alternatus. We surveyed its expression in various developmental stages of male and female adult tissues and determined its binding affinities for different pine volatiles using a competitive binding fluorescence assay. A CSP known as CSP5 in M. alternatus was obtained from an antennal cDNA library and expressed in Escherichia coli. Quantitative reverse transcription polymerase chain reaction results indicated that the CSP5 gene was mainly expressed in male and female antennae. Competitive binding assays were performed to test the binding affinity of recombinant CSP5 to 13 odour molecules of pine volatiles. The results showed that CSP5 showed very strong binding abilities to myrcene, (+)-β-pinene, and (−)-isolongifolene, whereas the volatiles 2-methoxy-4-vinylphenol, p-cymene, and (+)-limonene oxide have relatively weak binding affinity at pH 5.0. Three volatiles myrcene, (+)-β-pinene, and (−)-isolongifolene may play crucial roles in CSP5 binding with ligands, but this needs further study for confirmation. The sensitivity of insect to host plant volatiles can effectively be used to control and monitor the population through mass trapping as part of integrated pest management programs.

Keywords: olfactory-specific protein, volatiles, competitive binding assay, expression characteristics, qPCR

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Authors: Sung-Min Park, Chan Kwon Park, Chae-Bogk Kim

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The purpose of this study is to analyze the relationship between transaction fairness, social capital, supply chain integration and sustainability. Based on the previous studies, measurement items were determined by using SPSS 22 and exploratory factor analysis was performed, and again, using AMOS 21 for confirmatory factor analysis and path analysis was performed by using study items that satisfy reliability, validity, and appropriateness of measurement model. It has shown that transaction fairness has a (+) significant effect on social capital, social capital on supply chain integration, supply chain integration on economic sustainability and social sustainability, and has a (+), but not significant effect on environmental sustainability. It has shown that supply chain integration has been proven to play a role as a parameter between social capital and economic and social sustainability, but not as a parameter between environmental sustainability. Through this study, it is suggested that clearly examining the relationship between fairness of trade, social capital, supply chain integration and sustainability, maintaining fairness of the transaction make formation of social capital, and further integration of supply chain, and achieve sustainability of entire supply chain.

Keywords: transaction fairness, social capital, supply chain integration, sustainability

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Authors: D. A. Dakul, T. M. Akindigh, B. J. Dogonyaro, O. J. Abba, K. T. Tangtur, N. Sambo, J. A. E. Okopi, J. A. Yohanna, G. E. Imade, G. S. Mwansat, S. Oguche

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Malaria and urinary Schistosomaisis are both endemic in Nigeria and pose a serious health challenge in rural areas where co-infections are common. This descriptive cross sectional study was carried out to determine the prevalence of co-infection and the impact of concurrent infection on haemoglobin concentration, Eosinophil and CD4+ T-lymphocyte counts. Plasmodium falciparum and Schistosoma haematobium infection were determined by Malaria Rapid Diagnostic Test (MRDT) kits and the presence of visible haematuria respectively and confirmed by conventional Polymerase Chain Reaction (cPCR). P values < 0.05 were considered statistically significant. Of the 110 children examined, 13 (11.8%) had concurrent infection with Schistosoma haematobium falciparum, 46(41.8%) had Plasmodium falciparum infection while 16(14.5%) had Schistosoma haematobium infection. A strong association between co-infection and the ages of 10-15 years with a 36.4% prevalence of anaemia was observed. Malaria was significantly associated with anaemia than with concurrent infections or schistomiasis alone. Co-infection with both pathogens and a high prevalence of anaemia was observed in Jinduut community. Although the causes of anaemia are multi-factorial, further investigation into the extent to which malaria and urinary schistosomiasis contribute to anaemia is needed. Also, integrated control efforts must be strengthened to mitigate the impact of concurrent infection in this group of vulnerable members in the community. The results can be applied to other communities during control.

Keywords: co-Infection, plasmodium falciparum and scistosoma haematobium, Jinduut, Nigeria

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Authors: Amir Azaron

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In this research, a stochastic programming approach is developed for designing supply chains with uncertain parameters. Demands and selling prices of products at markets are considered as the uncertain parameters. The proposed mathematical model will be multi-period two-stage stochastic programming, which takes into account the selection of retailer sites, suppliers, production levels, inventory levels, transportation modes to be used for shipping goods, and shipping quantities among the entities of the supply chain network. The objective function is to maximize the chain’s net present value. In order to maximize the chain’s NPV, the sum of first-stage investment costs on retailers, and the expected second-stage processing, inventory-holding and transportation costs should be kept as low as possible over multiple periods. The effects of supply uncertainty where suppliers are unreliable will also be investigated on the efficiency of the supply chain.

Keywords: supply chain management, stochastic programming, multiobjective programming, inventory control

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Authors: Ude N. Callistus, Amulu F. Ndidi, Onukwuli D. Okechukwu, Amulu E. Patrick

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Power law approximation was used in this study to evaluate the reaction orders of calcium oxide, CaO catalyzed transesterification of refined cottonseed oil and methanol. The kinetics study was carried out at temperatures of 45, 55 and 65 ^oC. The kinetic parameters such as reaction order 2.02 and rate constant 2.8 hr^-1g^-1cat, obtained at the temperature of 65 ^oC best fitted the kinetic model. The activation energy, Ea obtained was 127.744 KJ/mol. The results indicate that the transesterification reaction of the refined cottonseed oil using calcium oxide catalyst is approximately second order reaction.

Keywords: refined cottonseed oil, transesterification, CaO, heterogeneous catalysts, kinetic model

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Authors: Neha Asthana, Vishal Krishna Prasad

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Reverse logistics and green supply chain form an interconnected and interwoven network of parameters that contribute to enhancement and incremental exchange in the triple bottom line in the consistently changing and fragmenting markets of the globalizing markets of today. Reverse logistics not only contributes to completing the supply chain in a comprehensive and synchronized manner but also contributes to a significant degree in optimizing green supply chains through procedures such as recycling, refurbishing etc. contributing to waste reduction. Carbon trading, owing to its limitations in the global context and being in a nascent stage seeks plethora of research to determine its full application in synergy with reverse logistics and green supply chain.

Keywords: reverse logistics, carbon trading, carbon emissions, green supply chain

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Authors: Nunthawan Nowwarote, Waleerat Sukarawan, Prasit Pavasant, Thanaphum Osathanon

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Interleukin 6 (IL-6) is a multifunctional cytokine, regulating various biological responses in several tissues. A Recent study shows that IL-6 plays a role in stemness maintenance in stem cells isolated from human exfoliated deciduous teeth (SHEDs). However, the role of IL-6 on cell differentiation in SHEDs remains unknown. The present study investigated the effect of IL-6 on SHEDs differentiation. Cells were isolated from dental pulp tissues of human deciduous teeth. Flow cytometry was used to determined mesenchymal stem cell marker expression, and the multipotential differentiation (osteogenic, adipogenic and neurogenic lineage ) was also determined. The mRNA was determined using real-time quantitative polymerase chain reaction, and the phenotypes were confirmed by chemical and immunofluorescence staining. Results demonstrated that SHEDs expressed CD44, CD73, CD90, CD105 but not CD45. Further, the up-regulation of osteogenic, adipogenic and neurogenic marker genes was observed upon maintaining cells in osteogenic, adipogenic and neurogenic induction medium, respectively. The addition of IL-6 induced osteogenic by up-regulated osteogenic marker gene also increased in vitro mineralization. Under neurogenic medium supplement with IL-6, up-regulated neurogenic marker. Whereas, an addition of IL-6 attenuated adipogenic differentiation by SHEDs. In conclusion, this evidence implies that IL-6 may participate in cells differentiation ability of SHEDs.

Keywords: SHEDs, IL-6, cell differentiations, dental pulp

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Authors: Fathilahbinti Ali, Jamarosliza Jamaluddin, Arun Kumar Upadhyay

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Poly(lactic acid) (PLA) is a biodegradable polymer which has good mechanical properties, however, its brittleness limits its usage especially in packaging materials. Therefore, in this work, PLA based polyurethane films were prepared by synthesizing with different types of isocyanates; methylene diisocyanate (MDI) and hexamethylene diisocyanates (HDI). For this purpose, PLA based polyurethane must have good strength and flexibility. Therefore, polycaprolactone which has better flexibility were prepared with PLA. An effective way to endow polylactic acid with toughness is through chain-extension reaction of the polylactic acid pre-polymer with polycaprolactone used as chain extender. Polyurethane prepared from MDI showed brittle behaviour, while, polyurethane prepared from HDI showed flexibility at same concentrations.

Keywords: biodegradable polymer, flexible, poly(lactic acid), polyurethane

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Authors: Anthony Ayodeji Adegoke, Olayinka Ayobami Aiyegoro, Thor Axel Stenstrom

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A study to investigate the species diversities, antibiogram and antibiotic resistance genes in Staphylococcus species from raw meat and dairy products collected from an abattoir and a farm shop of a research institute in Irene, South Africa over a six-month period was conducted. Polymerase Chain Reaction was used to speciate the bacteria and to detect the presence and otherwise of resistance genes. Antibiotic susceptibility testing was performed by disk diffusion method on Mueller-Hinton agar according to the Clinical Laboratory Standards Institute standards. A total of twenty-six (26) antibiotics were used to determine the antibiotic susceptibility. S. xylosus was the predominant isolate with 30% total occurrence, followed by S. epidermis, S. aureus, S. saprophyticus and S. haemolyticus with 25%, 15%, 15%, and 10% abundance respectively. The isolates were resistant to ceftezidime, gentamycin, nalidixic acid, nortrafuration, ampicillin, penicillin, oxytetracycline, tetracycline, doxycycline, clindamycin and lincomycin. mecA genes was detected among the methicillin resistant Staphylococcus species (MRSS) but no vancomycin resistance genes (van A and van B) were detected in these isolates. The presence of MRSS and multidrug resistant Staphylococcus species in meat affirms the need to avoid consumption of partially cooked meat currently rampant in South Africa, to avoid the spread of difficult to control pathogens in epidemiological proportion.

Keywords: Staphylococcus species, antibiotics, antibiotic resistance genes, food products, methicillin resistance, mecA gene

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Authors: Silvana Caglieri

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Theoretical study of acetylation of p-methylaniline catalyzed by Cu2+ ions from the analysis of intermediate of the reaction was carried out. The study of acetylation of amines is of great interest by the utility of its products of reaction and is one of the most frequently used transformations in organic synthesis as it provides an efficient and inexpensive means for protecting amino groups in a multistep synthetic process. Acetylation of amine is a nucleophilic substitution reaction. This reaction can be catalyzed by Lewis acid, metallic ion. In reaction mechanism, the metallic ion formed a complex with the oxygen of the acetic anhydride carbonyl, facilitating the polarization of the same and the successive addition of amine at the position to form a tetrahedral intermediate, determining step of the rate of the reaction. Experimental work agreed that this reaction takes place with the formation of a tetrahedral intermediate. In the present theoretical work were investigated the structure and energy of the tetrahedral intermediate of the reaction catalyzed by Cu2+ ions. Geometries of all species involved in the acetylation were made and identified. All of the geometry optimizations were performed by the method at the DFT/B3LYP level of theory and the method MP2. Were adopted the 6-31+G* basis sets. Energies were calculated using the Mechanics-UFF method. Following the same procedure it was identified the geometric parameters and energy of reaction intermediate. The calculations show 61.35 kcal/mol of energy for the tetrahedral intermediate and the energy of activation for the reaction was 15.55 kcal/mol.

Keywords: amides, amines, DFT, MP2

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Authors: Mojgan Zendehdel, Safura Molla Mohammad Zamani

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In this paper, Fe3O4/NaY zeolite nanocomposite with different molar ratio were successfully synthesized and characterized using FT-IR, XRD, TGA, SEM and VSM techniques. The SEM graphs showed that much of Fe3O4 was successfully coated by the NaY zeolite layer. Also, the results show that the magnetism of the products is stable with added zeolite. The catalytic effect of nanocomposite investigated for esterification reaction under solvent-free conditions. Hence, the effect of the catalyst amount, reaction time, reaction temperature and reusability of catalyst were considered and nanocomposite that created from zeolite and 16.6 percent of Fe3O4 showed the highest yield. The catalyst can be easily separated from reaction with the magnet and it can also be used for several times.

Keywords: zeolite, magnetic, nanocompsite, esterification

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Authors: Ergün Şakalar, Şeyma Özçirak Ergün

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Pistachio nuts, the fruits of the pistachio tree (Pistacia vera), are edible tree nuts highly valued for their organoleptic properties. Pistachio nuts used in snack foods, chocolates, baklava, meat products, ice-cream industries and other gourmet products as ingredients. Undeclared pistachios may be present in food products as a consequence of fraudulent substitution. Control of food samples is very important for safety and fraud. Mix of pistachio, peanut (Arachis hypogaea), pea (Pisum sativum L.) used instead of pistachio in food products, because pistachio is a considerably expensive nut. To solve this problem, a sensitive polymerase chain reaction PCR has been developed. A real-time PCR assay for the detection of pea, peanut and pistachio in baklava was designed by using EvaGreen fluorescence dye. Primers were selected from powerful regions for identification of pea, peanut and pistachio. DNA from reference samples and industrial products were successfully extracted with the GIDAGEN® Multi-Fast DNA Isolation Kit. Genomes were identified based on their specific melting peaks (Mp) which are 77°C, 85.5°C and 82.5°C for pea, peanut and pistachio, respectively. Homogenized mixtures of raw pistachio, pea and peanut were prepared with the ratio of 0.01%, 0.1%, 1%, 10%, 40% and 70% of pistachio. Quantitative detection limit of assay was 0.1% for pistachio. Also, real-time PCR technique used in this study allowed the qualitative detection of as little as 0.001% level of peanut DNA, 0,000001% level of pistachio DNA and 0.000001% level of pea DNA in the experimental admixtures. This assay represents a potentially valuable diagnostic method for detection of nut species adulterated with pistachio as well as for highly specific and relatively rapid detection of small amounts of pistachio in food samples.

Keywords: pea, peanut, pistachio, real-time PCR

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Authors: Adeleh Moshtaghi Zonouz, Issa Eskandari

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A simple, efficient, and green one-pot, four-component synthesis of highly substituted polyhydronaphthalenes in aqueous media is described. The method has such advantages as short reaction times, high yields, mild reaction conditions, operational simplicity and environmentally benign.

Keywords: polyhydronaphthalene, 2, 6-dicyanoanilines, multi-component reaction, aqueous media

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Authors: Zohreh Khojasteh-Ghamari, Takashi Irohara

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One of the efficient ways in supply chain risk management is avoiding the interruption in Supply Chain (SC) before it occurs. Although the majority of the organizations focus on their first-tier suppliers to avoid risk in the SC, studies show that in only 60 percent of the disruption cases the reason is first tier suppliers. In the 40 percent of the SC disruptions, the reason is downstream SC, which is the second tier and lower. Due to the increasing complexity and interrelation of modern supply chains, the SC elements have become difficult to trace. Moreover, studies show that there is a vital need to better understand the integration of risk and visibility, especially in the context of multiple objectives. In this study, we propose a multi-objective programming model to avoid disruption in SC. The objective of this study is evaluating the effect of downstream SCV on managing supply chain risk. We propose a multi-objective mathematical programming model with the objective functions of minimizing the total cost and maximizing the downstream supply chain visibility (SCV). The decision variable is supplier selection. We assume there are several manufacturers and several candidate suppliers. For each manufacturer, our model proposes the best suppliers with the lowest cost and maximum visibility in downstream supply chain. We examine the applicability of the model by numerical examples. We also define several scenarios for datasets and observe the tendency. The results show that minimum visibility in downstream SC is needed to have a safe SC network.

Keywords: downstream supply chain, optimization, supply chain risk, supply chain visibility

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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: Rym Ghariani, Ghada Soltane, Younes Boujelbene

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Industry 4.0, a set of emerging smart and digital technologies, has been the main focus of operations management researchers and practitioners in recent years. The objective of this research paper is to study the impact of Industry 4.0 on the integration of the supply chain (SCI) in Tunisian industrial companies. A conceptual model to study the relationship between Industry 4.0 technologies and supply chain integration was designed. This model contains three explained variables (Big data, Internet of Things, and Robotics) and one variable to be explained (supply chain integration). In order to answer our research questions and investigate the research hypotheses, principal component analysis and discriminant analysis were used using SPSS26 software. The results reveal that there is a statistically positive impact significant impact of Industry 4.0 (Big data, Internet of Things and Robotics) on the integration of the supply chain. Interestingly, big data has a greater positive impact on supply chain integration than the Internet of Things and robotics.

Keywords: industry 4.0 (I4.0), big data, internet of things, robotics, supply chain integration

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Authors: Sabriye Piskin, Sibel Kasap, Muge Sari Yilmaz

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Nanocrystalline TiO₂ particles were successfully synthesized via sol-gel and sonochemical combination using titanium tetraisopropoxide as a precursor at lower temperature for a short time. The effect of the reaction parameters (hydrolysis media, acid media, and reaction temperatures) on the synthesis of TiO₂ particles were investigated in the present study. Characterizations of synthesized samples were prepared by X-ray diffraction (XRD) analysis. It was shown that the reaction parameters played a significant role in the synthesis of TiO₂ particles.

Keywords: crystalline TiO2, sonochemical mechanism, sol-gel reaction, XRD

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Authors: Meenakshi Srivastava, A. K. Mishra

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This study characterizes the metabolic versatility of denitrifying bacterial communities residing in the paddy soil using the GC-MS based Phospholipid Fatty Acid (PLFA) analyses simultaneously with nosZ gene based PCR-DGGE (Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis) and real time Q-PCR analysis. We have analyzed the abundance of nitrous oxide reductase (nosZ) genes, which was subsequently related to soil PLFA profile and DGGE based denitrifier community structure. Soil denitrifying bacterial community comprised majority or dominance of Ochrobactrum sp. following Cupriavidus and uncultured bacteria strains in paddy soil of selected sites. Initially, we have analyzed the abundance of the nitrous oxide reductase gene (nosZ), which was found to be related with PLFA based lipid profile. Chandauli of Eastern UP, India represented greater amount of lipid content (C18-C20) and denitrifier’s diversity. This study suggests the positive co-relation between soil PLFA profiles, DGGE, and Q-PCR data. Thus, a close networking among metabolic abilities and taxonomic composition of soil microbial communities existed, and subsequently, such work at greater extent could be helpful in managing nutrient dynamics as well as microbial dynamics of paddy soil ecosystem.

Keywords: denaturing gradient gel electrophoresis, DGGE, nitrifying and denitrifying bacteria, PLFA, Q-PCR

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Authors: Hend K. Moosa, Eman A. Rashwan, Ezzat M. Hassan, Amany A. Ghazy, Amel G. Sheredy

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The stability of microRNA (miR) in the circulation can show a great progress toward the discovery of non-invasive diagnostic and prognostic biomarkers in many diseases. In the present study, circulatory miR-122 and miR-130a were analysed in chronic hepatitis C Egyptian patients in predicting the clinical outcome of interferon treatment. In addition, their expression levels were correlated to viral RNA levels, necro-inflammatory markers (AST, ALT) and to each other. This study was conducted on 51 subjects where 36 were chronic HCV patients in which they were divided into naive and interferon treated HCV patients (responders and non-responders) and 15 matched healthy controls. Serum quantification of miR-122 and miR-130a were performed by quantitative Real-time Polymerase Chain Reaction (qRT-PCR). The results showed a significant upregulation of miR-122 in non-responder patients (P=0.049). By receiver operating characteristic analysis curve, miR-122 revealed 65% sensitivity and 92.3% specificity in predicting non-responsiveness of patients to IFN treatment, while miR-130a showed a sensitivity of 100% and specificity of 53.85%. Remarkably, there was a significant positive correlation between miR-122 and miR-130a in naive HCV patients (r=0.714, p=0.003). However, there was no significant correlation between serum miR-122, miR-130a expression levels and necro-inflammatory markers (AST, ALT). To conclude, miR-122 and miR-130a have a significant association with viral RNA levels and accordingly, they may have a synergistic power in promoting viral replication. Interestingly, miR-122 and miR-130a have a predictive power in predicting clinical outcome of IFN treatment which can be further studied in currently used drugs in order to reduce the socio-economic burden of potentially non-responders.

Keywords: hepatitis C, microRNA, miR-122, miR-130a

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Authors: Samir Hmaimou, Marouane Ait Lahcen, Mohamed Adardour, Mohamed Maatallah, Abdesselam Baouid

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Due to (3 + 2) cycloaddition and condensation reaction, a wide range of synthetic routes can be used to obtain biologically active heterocyclic compounds. Condensation and (3+2) cycloaddition reactions in heterocyclic syntheses are versatile due to the wide variety of possible combinations of several atoms of the reactants. In this article, we first outline the synthesis of benzodiazepine 4 with two dipolarophilic centers (C=C and C=N) by condensation reaction. Then, we use it for cycloaddition reactions (3+2) with nitrile oxides to prepare oxadiazole-benzodiazepines and pyrazole-benzodiazepine compounds. ¹H and ¹³C NMR are used to establish all the structures of the synthesized products. These condensation and cycloaddition reactions were then analyzed using density functional theory (DFT) calculations at the B3LYP/6-311G(d,p) theoretical level. In this study, the mechanism of the one-step cycloaddition reaction was investigated. Molecular electrostatic potential (MEP) was used to identify the electrophilic and nucleophilic attack sites of the molecules studied. Additionally, Fukui investigations (electrophilic f- and nucleophilic f+) in the various reaction centers of the reactants demonstrate that, whether in the condensation reaction or cycloaddition, the reaction proceeds through the atomic centers with the most important Fukui functions, which is in full agreement with experimental observations. In the condensation reaction, thermodynamic control of regio, chemo, and stereoselectivity is observed, while those of cycloaddition are subject to kinetic control.

Keywords: cycloaddition reaction, regioselectivity, mechanism reaction, NMR analysis

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Authors: Yue Meng, Lingyun Wei

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In this paper, the coordination problem of a supply chain system composed of multiple retailers and manufacturers is studied under the background of the new retail supply chain. Taking a system composed of two retailers and one manufacturer as an example, this paper introduces an online store owned by the manufacturer to reflect the characteristics of the combination of online and offline new retail. Then, this paper gives the conditions that need to be satisfied to realize the coordination between retailers and manufacturers, such as the revenue sharing coefficient. The supply chain coordination model is compared with the newsboy model through a specific example. Finally, the conclusion is drawn that the profits of the coordinated supply chain and its members are better than the corresponding profits under the newsboy model; that is, the coordination of the supply chain is realized by using the revenue sharing contract and the transshipment fund mechanism.

Keywords: transshipment, coordination, multi-retailer, revenue-sharing contract

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Authors: M. R. Haji Hajikolaei, A. A. Nikvand, A. R. Ghadrdan, M. Ghorbanpoor, B. Mohammadian

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This study was carried out on water buffalo for detection of Leptospira interrogans in kidney and urine and its relationship with serological findings. Blood, urine and kidney samples were taken immediately after slaughter from 353 water buffalos at Ahvaz abattoir in Khouzestan province, Iran. Sera were initially screened at serum dilution of 1:100 against seven live antigens of Leptospira interrogans: pomona, hardjo, ballum, icterohemorrhagiae, tarasovi, australis and grippotyphosa using the microscopic agglutination test (MAT) and sera with positive results were titrated against reacting antigens in serial twofold dilution from 1:100 to 1:800. The samples of kidney were embedded in paraffin wax and 5µm thick sections were stained routinely with Haematoxylin and Eosin (H&E). Polymerase chain reaction (PCR) examination was done on urine and kidney by using LipL32 gene primers. Antibodies against one or more serovars at dilution >:100 were detected in sera. The most frequent reactor was hardjo (56.2%), followed by pomona (52.3%), australis (9.8%), tarassovi (5.9%), grippotyphosa (4.5%) and icterohaemorrhagiae (3.9%). The L. interrogans were detected in 43 (12.2%) of examined buffaloes, so that 26 (8.2%) of kidney tissues, 14 (4.8%) of urine samples separately and 3 (0.84%) of both kidney and urine samples were positive in PCR. From 153 (43.3%) buffaloes with positive MAT, 24 cases were positive by PCR of kidney and/or urine samples, synchronously. Renal lesions such as interstitial nephritis, acute tubular necrosis (ATN), pyelonephritis, glomerolonephritis, renal fibrosis and hydronephrosis were found in 128 (36.3%) cases. Statistical analysis indicated that there was no significant association between results of MAT, PCR and interstitial nephritis.

Keywords: leptospiral infection, PCR, MAT, histopathology, river buffalo

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Authors: Andreas Latz, Daniela Heinz, Fatima Hashemi, Melek Baygül

Abstract:

Introduction: Feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), and feline coronavirus (FCoV) are serious infectious diseases affecting cats worldwide. Transmission of these viruses occurs primarily through close contact with infected cats (via saliva, nasal secretions, faeces, etc.). FeLV, FIV, and FCoV infections can occur in combination and are expressed in similar clinical symptoms. Diagnosis can therefore be challenging: Symptoms are variable and often non-specific. Sick cats show very similar clinical symptoms: apathy, anorexia, fever, immunodeficiency syndrome, anemia, etc. Sample volume for small companion animals for diagnostic purposes can be challenging to collect. In addition, multiplex diagnosis of diseases can contribute to an easier, cheaper, and faster workflow in the lab as well as to the better differential diagnosis of diseases. For this reason, we wanted to develop a new diagnostic tool that utilizes less sample volume, reagents, and consumables than multiplesingleplex ELISA assays Methods: The Multiplier from Dynextechonogies (USA) has been used as platform to develop a Multiplex diagnostic tool for the detection of antibodies against FIV and FCoV/FIP and antigens for FeLV. Multiplex diagnostics. The Dynex®Multiplier®is a fully automated chemiluminescence immunoassay analyzer that significantly simplifies laboratory workflow. The Multiplier®ease-of-use reduces pre-analytical steps by combining the power of efficiently multiplexing multiple assays with the simplicity of automated microplate processing. Plastic beads have been coated with antigens for FIV and FCoV/FIP, as well as antibodies for FeLV. Feline blood samples are incubated with the beads. Read out of results is performed via chemiluminescence Results: Bead coating was optimized for each individual antigen or capture antibody and then combined in the multiplex diagnostic tool. HRP: Antibody conjugates for FIV and FCoV antibodies, as well as detection antibodies for FeLV antigen, have been adjusted and mixed. 3 individual prototyple batches of the assay have been produced. We analyzed for each disease 50 well defined positive and negative samples. Results show an excellent diagnostic performance of the simultaneous detection of antibodies or antigens against these feline diseases in a fully automated system. A 100% concordance with singleplex methods like ELISA or IFA can be observed. Intra- and Inter-Assays showed a high precision of the test with CV values below 10% for each individual bead. Accelerated stability testing indicate a shelf life of at least 1 year. Conclusion: The new tool can be used for multiplex diagnostics of the most important feline infectious diseases. Only a very small sample volume is required. Fully automation results in a very convenient and fast method for diagnosing animal diseases.With its large specimen capacity to process over 576 samples per 8-hours shift and provide up to 3,456 results, very high laboratory productivity and reagent savings can be achieved.

Keywords: Multiplex, FIV, FeLV, FCoV, FIP

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Authors: Ramon Erasmo Verdin Rodriguez

Abstract:

Many has been the ways that historically the managers and gurus has been trying to get closer to the perfect supply chain, but since this topic is so vast and very complex the bigger the companies are, the duty has not been certainly easy. On this research, you are going to see thru the entrails of the logistics that happens at a glass manufacturing company with the number one raw material of the process that is the silica sand. After a very quick passage thru the supply chain, this document is going to focus on the way that raw materials flow thru the system, so after that, an analysis and research can take place to improve the logistics. Thru Operations Research techniques, it will be analyzed the current scheme of distribution and inventories of raw materials at a glass company’s plants, so after a mathematical conceptualization process, the supply chain could be optimized with the purpose of reducing the uncertainty of supply and obtaining an economic benefit at the very end of this research.

Keywords: inventory management, operations research, optimization, supply chain

Procedia PDF Downloads 326

Authors: Iddrisu Ibrahim, Joseph Atia Ayariga, Junhuan Xu, Daniel Abugri, Boakai Robertson, Olufemi S. Ajayi

Abstract:

The emergence of multidrug resistance poses a huge risk to public health globally. Yet these recalcitrant pathogens continue to rise in incidence rate, with resistance rates significantly outpacing the speed of antibiotic development. This, therefore, presents an aura of related health issues such as untreatable nosocomial infections arising from organ transplants and surgeries, as well as community-acquired infections that are related to people with compromised immunity, e.g., diabetic and HIV patients, etc. There is a global effort to fight multidrug-resistant pathogens spearheaded by the World Health Organization, thus calling for research into novel antimicrobial agents to fight multiple drug resistance. Previously, our laboratory demonstrated that Cannabidiol (CBD) was an effective antimicrobial against Salmonella typhimurium (S. typhimurium). However, we observed resistance development over time. To understand the mechanisms S. typhimurium uses to develop resistance to Cannabidiol (CBD), we studied the abundance of bacteria lipopolysaccharide (LPS) and membrane sterols of both susceptible and resistant S. typhimurium. Using real-time quantitative polymerase chain reaction (RT-qPCR), we also analyzed the expression of selected genes known for aiding resistance development in S. typhimurium. We discovered that there was a significantly higher expression of blaTEM, fimA, fimZ, and integrons in the CBD-resistant bacteria, and these were also accompanied by a shift in abundance in cell surface molecules such as lipopolysaccharide (LPS) and sterols.

Keywords: antimicrobials, resistance, cannabidiol, gram-negative bacteria, integrons, blaTEM, Fim, LPS, ergosterols

Procedia PDF Downloads 101

Authors: Jung-Hsuan Hsu

Abstract:

Due to intense competition within the industry, companies have increasingly recognized partnerships with other companies. In addition, with outsourcing and globalization of the supply chain, it leads to companies' increasing reliance on external resources. Consequently, supply chain network becomes complex, so that it reduces the visibility of the manufacturing process. Therefore, this study is going to focus on the impact of supply chain relationship quality (SCRQ) on cooperative strategy and visibility. Questionnaire survey is going to be conducted as research method, using the organic food industry as the research subject, and the sampling method is random sampling. Finally, the data analysis will use SPSS statistical software and AMOS software to analyze and verify the hypothesis. The expected results in this study is to evaluate the supply chain relationship quality between Taiwan's food manufacturing and their suppliers regarding whether it has a positive impact for the persistence, frequency and diversity of cooperative strategy, as well as the dimensions of supply chain relationship quality on visibility regarding whether it has a positive effect.

Keywords: supply chain relationship quality (SCRQ), cooperative strategy, visibility, competition

Procedia PDF Downloads 451

Authors: Hung Nguyen

Abstract:

With the rise of global supply chain network, the choice of supply chain orientation is critical. The alignment between cultural similarity and supply chain information exchange could help identify appropriate supply chain orientations, which would differentiate the stronger competitors and performers from the weaker ones. Through developing a taxonomy, this study examined whether the choices of action programs and manufacturing performance differ depending on the levels of attainment cultural similarity and information exchange. This study employed statistical tests on a large-scale dataset consisting of 680 manufacturing plants from various cultures and industries. Firms need to align cultural practices with the level of information exchange in order to achieve good overall business performance. There appeared to be consistent three major orientations: the Proactive, the Initiative and the Reactive. Firms are experiencing higher payoffs from various improvements are the ones successful alignment in both information exchange and cultural similarity The findings provide step-by-step decision making for supply chain information exchange and offer guidance especially for global supply chain managers. In including both cultural similarity and information exchange, this paper adds greater comprehensiveness and richness to the supply chain literature.

Keywords: culture, information exchange, supply chain orientation, similarity

Procedia PDF Downloads 359