Search results for: quantitative polymerase chain reaction
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 7172

Search results for: quantitative polymerase chain reaction

6992 Study of the Genes Involved in the Resistance of Nosocomial Pseudomonas aeruginosa to Fluoroquinolone

Authors: Rosetta Moshirian Farahi, Ahya Abdi Ali, Sara Gharavi

Abstract:

The major mechanism of Pseudomonas aeruginosa resistance to fluoroquinolones is the alteration of target enzymes, type II and IV topoisomerases due to mutations in the quinolone resistance-determining regions (QRDR) of the gyrA and parC genes coding A subunits of these enzymes. 37 isolates from patients with burn wounds and 20 isolates from blood, urine and sputum specimen were selected to evaluate mutations involved in antibiotic resistance and were subsequently verified for their resistance to ciprofloxacin. QRDRs regions of gyrA and parC were amplified by polymerase chain reaction (PCR) and were subsequently sequenced. 90% of isolates with MIC≥8 µg/ml to ciprofloxacin had a mutation in gyrA gene in which threonine at position 83 changed to isoleucine. 87.5% of isolates had mutation in parC, Serine 87 changed. 75% had Ser87Leu and 12.5% possessed Serin87Trp. Various silent mutations were also detected such as Val103Val, Ala118Ala, Ala136Ala, His132His in gyrA and Ala115Ala in parC. The data indicates that the common mutation in gyrA is Thr83Ile and in parC is Ser87Leu/Trp. No individual parC mutation was observed while mutations in gyrA and parC occurred simultaneously and appears to be the main reason of high-level resistance to fluoroquinolones in patients with burn wounds and urine infection. The vast majority of P.aeruginosa isolates had mutation in parC which can play a crucial role in increased resistance of these isolates. This is a report of parC mutations from resistant P. aeruginosa isolates from Iran, Tehran.

Keywords: P. aeruginosa, fluoroquinolones, gyrA, parC, antibiotic resistance

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6991 Molecular Detection of Viruses Causing Hemorrhagic Fevers in Rodents in the South-West of Korea

Authors: Sehrish Jalal, Choon-Mee Kim, Dong-Min Kim

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Background: Many pathogens causing hemorrhagic fevers of medical and veterinary importance have been identified and isolated from rodents in the Republic of Korea (ROK). Objective: We investigated the prevalence of emerging viruses causing hemorrhagic fevers, such as hemorrhagic fever with renal syndrome (HFRS), severe fever with thrombocytopenia syndrome (SFTS) and flaviviruses, from wild rodents. Methods: Striped field mice, Apodemus agrarius, (n=39) were captured during 2014-2015 in the south-west of ROK. Using molecular methods, lung samples were evaluated for SFTS virus, HFRS virus and flavivirus, and seropositivity was evaluated in the blood. Results: A high positive rate of Hantavirus (46.2%) was detected in A.agrarius lungs by reverse transcription-nested polymerase chain reaction (RT-N-PCR). The monthly prevalence of HFRS virus was 16.7% in October, 86.7% in November and 25% in August of the following year (p < 0.001). Moreover, 17.9% of blood samples were serologically positive for Hantavirus antibodies. The most prevalent strain in A. agrarius was Hantaan virus. All samples were positive for neither SFTS nor flavivirus. Conclusion: Hantan virus was detected in 86.7% of A. agrarius in November (autumn), and thus, virus shedding from A. agrarius can increase the risk of humans contracting HFRS. These findings may help to predict and prevent disease outbreaks in ROK.

Keywords: hemorrhagic fever virus, molecular diagnostic technique, rodents, Korea

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6990 Pathological and Molecular Diagnosis of Caseous Lymphadenitis in Chinkara Deer (Gazella Bennettii), in Pakistan

Authors: Mudassar Iqbal, Riaz Hussain, Khalid Mehmood, Farah Ali, Fazal Mahmood, Abdul Ghaffar

Abstract:

Corynebacterium pseudotuberculosis is an important cause of caseous lymphadenitis (CL), a complex, chronic devastating and destructive disease of small ruminants. In present study, postmortem examination of Chinkara deer (n=25) was conducted in year 2014. Pus samples suggestive of CL were collected from the superficial lymph nodes, liver, spleen and lungs during necropsy and subjected to standard microbiological procedures for isolation and molecular analysis of bacterial pathogens. Pus samples collected from carcasses (25) presenting clinical lesions of C. pseudotuberculosis infection was identified in 19 (76%) carcasses on the basis of culture characteristics. The frequency of C. pseudotuberculosis bacterium was higher in older animals as compared to young animals. Grossly, multiple tubercles of variable size having caseous material were observed in liver, lungs, spleen and lymph nodes. Histopathologically, tissue sections from all the visceral organs were extensively plugged with abscess. In present study specific prolineiminopeptidase (PIP) gene of the C. pseudotuberculosis was amplified by the Polymerase chain reaction technique (PCR) in 17(25) cases. The efficient and reliable molecular analysis along with necropsy findings in present study can be used as valuable approach for diagnosis of caseous lymphadenitis in small ruminants.

Keywords: Chinkara deer, Corynebacterium pseudotuberculosis, Caseous lymphadenitis, PCR

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6989 Selection of Suitable Reference Genes for Assessing Endurance Related Traits in a Native Pony Breed of Zanskar at High Altitude

Authors: Prince Vivek, Vijay K. Bharti, Manishi Mukesh, Ankita Sharma, Om Prakash Chaurasia, Bhuvnesh Kumar

Abstract:

High performance of endurance in equid requires adaptive changes involving physio-biochemical, and molecular responses in an attempt to regain homeostasis. We hypothesized that the identification of the suitable reference genes might be considered for assessing of endurance related traits in pony at high altitude and may ensure for individuals struggling to potent endurance trait in ponies at high altitude. A total of 12 mares of ponies, Zanskar breed, were divided into three groups, group-A (without load), group-B, (60 Kg) and group-C (80 Kg) on backpack loads were subjected to a load carry protocol, on a steep climb of 4 km uphill, and of gravel, uneven rocky surface track at an altitude of 3292 m to 3500 m (endpoint). Blood was collected before and immediately after the load carry on sodium heparin anticoagulant, and the peripheral blood mononuclear cell was separated for total RNA isolation and thereafter cDNA synthesis. Real time-PCR reactions were carried out to evaluate the mRNAs expression profile of a panel of putative internal control genes (ICGs), related to different functional classes, namely glyceraldehyde 3-phosphate dehydrogenase (GAPDH), β₂ microglobulin (β₂M), β-actin (ACTB), ribosomal protein 18 (RS18), hypoxanthine-guanine phosophoribosyltransferase (HPRT), ubiquitin B (UBB), ribosomal protein L32 (RPL32), transferrin receptor protein (TFRC), succinate dehydrogenase complex subunit A (SDHA) for normalizing the real-time quantitative polymerase chain reaction (qPCR) data of native pony’s. Three different algorithms, geNorm, NormFinder, and BestKeeper software, were used to evaluate the stability of reference genes. The result showed that GAPDH was best stable gene and stability value for the best combination of two genes was observed TFRC and β₂M. In conclusion, the geometric mean of GAPDH, TFRC and β₂M might be used for accurate normalization of transcriptional data for assessing endurance related traits in Zanskar ponies during load carrying.

Keywords: endurance exercise, ubiquitin B (UBB), β₂ microglobulin (β₂M), high altitude, Zanskar ponies, reference gene

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6988 Prevalence of High Risk Human Papillomavirus in Cervical Dysplasia and Cancer Samples from Twin Cities in Pakistan

Authors: Sana Gul, Sheeba Murad, Aneela Javed

Abstract:

Introduction: Human Papilloma Virus (HPV) is small DNA virus mostly infecting mucosa and cutaneous keratinocytes. So far, more than 200 Human papillomaviruses are known. HPV have been divided into high- and low-risk on the basis of their oncogenic potential. High risk HPV is considered to be the main etiological cause for cervical cancer. Objective: Current study was designed to screen the local cervical cancer patients from the twin cities of Pakistan for the occurance of high risk HPV. Methodology: A total of 67 formalin fixed paraffin-embedded samples of cervical cancer biopsies were obtained from the government hospitals in Islamabad and Rawalpindi. Cervical cancer biopsies were examined for the presence of HPV DNA. Polymerase chain reaction (PCR) was used for the amplification of a region in the HPV-L1 gene for the general detection of the Papilloma virus and for the genotype specific detection of high risk HPV 16 and 18 using the GP5/GP6 primers and genotype specific primers respectively. Results: HPV DNA was detected in 59 out of 67 samples analyzed. 30 samples showed the presence of HPV16 while 22 samples were positive for HPV 18 . HPV subtype could not be determined in 7 samples. Conclusion: Our results show a strong association between HPV infection and cervical cancer among women in twin cities of Pakistan. One way to minimize the disease burden in relation to HPV infection in Pakistani population is the use of prophylactic vaccines and routine screening. An early diagnosis of HPV infection will allow better health management to reduce the risk of developing cervical cancer.

Keywords: cervical cancer, Pakistan, human papillomavirus, HPV 16

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6987 Investigation of the Composition and Structure of Tar by Lignite Pyrolysis Using Thermogravimetry, Gas Chromatography and Mass Spectrum Coupled Instrument System

Authors: Li Feng, Cheng Zhang, Chuanzhou Yuang

Abstract:

Understanding the macromolecular structure of low-rank coal is very important for its gasification and liquefaction. The pyrolysis is one of the methods of analyzing the macromolecular structure of coal. The gaseous products decomposed directly by the raw lignite at 500 °C and indirectly by tar products from raw lignite pyrolysis at 500 °C were investigated and compared by thermogravimetry, gas chromatography and mass spectrum coupled instrument system (TG/GC/MS) in this paper. The results show that 52 kinds of products were found from the raw lignite and 70 kinds of products from the tar. The pyrolysis products directly from the lignite appear more monocyclic aromatic hydrocarbons and less substituent groups or branch chain, compared with the products from the tar. There is less linear chain and double bonds structure in the tar, which can be speculated that linear chain and double bonds structure took part in the generation of condensed rings and other reactions. There are more kinds of phenol and furan in the tar, which indicate that these products may be generated from the secondary reaction. The formation process of phenol, phenol naphthalene, naphthene and furan are discussed.

Keywords: composition and structure, lignite, pyrolysis of coal, tar, TG/GC/MS

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6986 A C/T Polymorphism at the 5’ Untranslated Region of CD40 Gene in Patients Associated with Graves’ Disease in Kumaon Region

Authors: Sanjeev Kumar Shukla, Govind Singh, Prabhat Pant Shahzad Ahmad

Abstract:

Background: Graves’ disease is an autoimmune disorder with a genetic predisposition, and CD40 plays a pathogenic role in various autoimmune diseases. A single nucleotide polymorphism at position –1 of the Kozak sequence of the 5 untranslated regions of the CD40 gene of exon 1 has been reported to be associated with the development of Graves’ Disease. Objective: The aim of the present study was to investigate whether CD40 gene polymorphism confers susceptibility to Graves’ disease in the Kumaon region. CD40 gene polymorphisms were studied in Graves’ Disease patients (n=50) and healthy control subjects without anti-thyroid autoantibodies or a family history of autoimmune disorders (n=50). Material and Method: CD40 gene polymorphisms were studied in fifty Graves’ Disease patients and fifty healthy control subjects. All samples were collected from STG Hospital, Haldwani, Nainital. A C/T polymorphism at position –1 of the CD40 gene was measured using the polymerase chain reaction-restriction fragment length polymorphism. Results: There was no significant difference in allele or genotype frequency of the CD40 SNP between Graves’ Disease and control subjects. There was a significant decrease in the TT genotype frequency in the Graves’ Disease patients who developed Graves’ Disease after 40 years old than those under 40 years of age. These data suggest that the SNP of the CD40 gene is associated with susceptibility to the later onset of Graves’ Disease. Conclusion: The CD40 gene was a different susceptibility gene for Graves’ Disease within certain families because it was both linked and associated with Graves’ Disease.

Keywords: autoimmune diseases, pathogenesis, diagnosis, therapy

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6985 Supply Chain Optimisation through Geographical Network Modeling

Authors: Cyrillus Prabandana

Abstract:

Supply chain optimisation requires multiple factors as consideration or constraints. These factors are including but not limited to demand forecasting, raw material fulfilment, production capacity, inventory level, facilities locations, transportation means, and manpower availability. By knowing all manageable factors involved and assuming the uncertainty with pre-defined percentage factors, an integrated supply chain model could be developed to manage various business scenarios. This paper analyse the utilisation of geographical point of view to develop an integrated supply chain network model to optimise the distribution of finished product appropriately according to forecasted demand and available supply. The supply chain optimisation model shows that small change in one supply chain constraint is possible to largely impact other constraints, and the new information from the model should be able to support the decision making process. The model was focused on three areas, i.e. raw material fulfilment, production capacity and finished products transportation. To validate the model suitability, it was implemented in a project aimed to optimise the concrete supply chain in a mining location. The high level of operations complexity and involvement of multiple stakeholders in the concrete supply chain is believed to be sufficient to give the illustration of the larger scope. The implementation of this geographical supply chain network modeling resulted an optimised concrete supply chain from raw material fulfilment until finished products distribution to each customer, which indicated by lower percentage of missed concrete order fulfilment to customer.

Keywords: decision making, geographical supply chain modeling, supply chain optimisation, supply chain

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6984 RNA Antisense Coat Protein Showing Promising Effects against Cotton Leaf Curl Disease in Pakistani Cotton

Authors: Zunnu Raen Akhtar

Abstract:

Cotton Leaf Curl Disease (CLCuD) is from Gemini virus and is transmitted through whiteflies in cotton. Transgenic cotton containing Antisense Coat Protein (ACP) has been found to show better results against CLCuD in cotton. In current research, Antisense Coat Protein was inserted in cotton plants to observe resistance developed in the cotton plants against CLCuD. T1 generation of plants were observed for its expression in plants. Tests were carried out to observe the expression of Antisense Coat Protein using Polymerase Chain Reaction (PCR) technique and by southern blotting. Whiteflies showing positive Cotton Leaf Curl Virus (CLCV) were reared and released in bioassay on ACP expressing cotton plants under laboratory as well as confined semi-field conditions. Results confirmed the expression of AC protein in PCR and southern blotting. Further laboratory results showed that cotton plants expressing AC protein showed rare incidence of CLCuD infection as compared to control. In the confined semi-field, similar results were observed in AC protein expressing cotton as compared to control. These results explicitly show that ACP can help to tackle the CLCuD issue in the future and further studies on biochemical processes involved in these plants and effects of ACP induction on non-target organisms should also be studied for eco-system.

Keywords: cotton, white flies, antisense coat protein, CLCV

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6983 Impact of a Locally-Prepared Fermented Alcoholic Beverage from Jaggery on the Gut Bacterial Profile of the Tea-Tribal Populations of Assam, India

Authors: Rupamoni Thakur, Madhusmita Dehingia, Narayan C. Talukdar, Mojibur R. Khan

Abstract:

The human gut is an extremely active fermentation site and is inhabited by diverse bacterial species. Consumption of alcoholic beverages has been shown to substantially modulate the human gut bacterial profile (GBP) of an individual. Assam, a major north-eastern state of India, is home to a number of tribal populations of which the tea-tribes form a major community. These tea-tribal communities are known to prepare and consume a locally-prepared alcoholic beverage from fermented jaggery, whose chemical composition is unknown. In this study, we demonstrate the effect of daily intake of the locally-prepared alcoholic beverage on the GBP of the tea-tribal communities and correlate it with the changes in the biochemical biomarkers of the population. The fecal bacterial diversity of 40 drinkers and 35 non-drinking healthy individuals were analyzed by polymerase chain reaction (PCR)–denaturing gradient gel electrophoresis (DGGE). The results suggested that the GBP was significantly modulated in the fermented-beverage consuming subjects. Significant difference was also observed in the serum biochemical parameters such as triglyceride, total cholesterol and the liver marker enzymes (ASAT/ALAT and GGT). Further studies to identify the GBP of drinkers vs non-drinkers through Next-generation Sequencing (NGS) analysis and to correlate the changes with the biochemical biomarkers of the population is underway.

Keywords: alcoholic beverage, gut bacterial profile, PCR-DGGE analysis, tea-tribes of India

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6982 Bio-Genetic Activities Associated with Resistant in Peppers to Phytophthora capsici

Authors: Mehdi Nasr-Esfahani, Leila Mohammad Bagheri, Ava Nasr-Esfahani

Abstract:

Root and collar rot disease caused by Phytophthora capsici (Leonian) is one of the most serious diseases in pepper, Capsicum annuum L. In this study, a diverse collection of 37 commercial edible and ornamental pepper genotypes infected with P. capsici were investigated for biomass parameters and enzymatic activity of peroxidase or peroxide reductases (EC), superoxide dismutase (SOD), polyphenol oxidase (PPOs), catalase (CAT) and phenylalanine ammonia-lyase (PAL). Seven candidate DEG genes were also evaluated on resistant and susceptible pepper cultivars, through measuring product formation, using spectrophotometry and real-time polymerase chain reaction. All the five enzymes and seven defense-gene candidates were up-regulated in all inoculated pepper accessions to P. capsici. But, the enzymes and DEG genes were highly expressed in resistant cv. 19OrnP-PBI, 37ChillP-Paleo, and “23CherryP-Orsh". The expression level of enzymes were 1.5 to 5.6-fold higher in the resistant peppers, than the control non-inoculated genotypes. Also, the transcriptional levels of related candidate DEG genes were 3.16 to 5.90-fold higher in the resistant genotypes. There was a direct and high correlation coefficient between resistance, bio-mass parameters, enzymatic activity, and resistance gene expression. The related enzymes and candidate genes expressed herein will provide a basis for further gene cloning and functional verification studies, and also will aid in an understanding of the regulatory mechanism of pepper resistance to P. capsici.

Keywords: AP2/ERF, cDNA, enzymes, MIP gene, q-RTPCR, XLOC

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6981 Investigation of Rifampicin and Isoniazid Resistance Mutated Genes in Mycobacterium Tuberculosis Isolated From Patients

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

Abstract:

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

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

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6980 Modified Gold Screen Printed Electrode with Ruthenium Complex for Selective Detection of Porcine DNA

Authors: Siti Aishah Hasbullah

Abstract:

Studies on identification of pork content in food have grown rapidly to meet the Halal food standard in Malaysia. The used mitochondria DNA (mtDNA) approaches for the identification of pig species is thought to be the most precise marker due to the mtDNA genes are present in thousands of copies per cell, the large variability of mtDNA. The standard method commonly used for DNA detection is based on polymerase chain reaction (PCR) method combined with gel electrophoresis but has major drawback. Its major drawbacks are laborious, need longer time and toxic to handle. Therefore, the need for simplicity and fast assay of DNA is vital and has triggered us to develop DNA biosensors for porcine DNA detection. Therefore, the aim of this project is to develop electrochemical DNA biosensor based on ruthenium (II) complex, [Ru(bpy)2(p-PIP)]2+ as DNA hybridization label. The interaction of DNA and [Ru(bpy)2(p-HPIP)]2+ will be studied by electrochemical transduction using Gold Screen-Printed Electrode (GSPE) modified with gold nanoparticles (AuNPs) and succinimide acrylic microspheres. The electrochemical detection by redox active ruthenium (II) complex was measured by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The results indicate that the interaction of [Ru(bpy)2(PIP)]2+ with hybridization complementary DNA has higher response compared to single-stranded and mismatch complementary DNA. Under optimized condition, this porcine DNA biosensor incorporated modified GSPE shows good linear range towards porcine DNA.

Keywords: gold, screen printed electrode, ruthenium, porcine DNA

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6979 Bi-Objective Optimization for Sustainable Supply Chain Network Design in Omnichannel

Authors: Veerpaul Maan, Gaurav Mishra

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The evolution of omnichannel has revolutionized the supply chain of the organizations by enhancing customer shopping experience. For these organizations need to develop well-integrated multiple distribution channels to leverage the benefits of omnichannel. To adopt an omnichannel system in the supply chain has resulted in structuring and reconfiguring the practices of the traditional supply chain distribution network. In this paper a multiple distribution supply chain network (MDSCN) have been proposed which integrates online giants with a local retailers distribution network in uncertain environment followed by sustainability. To incorporate sustainability, an additional objective function is added to reduce the carbon content through minimizing the travel distance of the product. Through this proposed model, customers are free to access product and services as per their choice of channels which increases their convenience, reach and satisfaction. Further, a numerical illustration is being shown along with interpretation of results to validate the proposed model.

Keywords: sustainable supply chain network, omnichannel, multiple distribution supply chain network, integrate multiple distribution channels

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6978 Short-Term Exposing Effects of 4,4'-DDT on Mitochondrial Electron Transport Complexes in Eyes of Zebrafish

Authors: Eun Ko, Moonsung Choi, Sooim Shin

Abstract:

4,4’-Dichlorodiphenyltrichloroethane (4,4’-DDT) is colorless, odorless organochlorine and known as persistent toxic organic pollutant accumulated in organs. In this study, effects of 4,4’-DDT on activities of mitochondrial electron transport chain system was analyzed. 4,4’-DDT is directly treated to isolated mitochondria from eyes of zebrafish and then activities of mitochondrial complex I, II, III, IV were measured spectrophotometrically. The reaction was proceeded immediately after adding 4,4’-DDT to examine the short-term exposing effects of persistent organic pollutant. As a result, high concentration of 4,4’-DDT treated mitochondria exhibited slightly enhanced activity in all complexes than non-treated one except complex III in male. Particularly, 4,4’-DDT was more effective on enzymatic activity in mitochondria isolated from eyes of male zebrafish. These results represented that 4,4’-DDT might temporarily induce to open up ion channel on isolated mitochondria resulting in increasing the functional activity of mitochondrial electron transport chain system.

Keywords: electron transport chain, mitochondrial function, persistent organic pollutant, spectrophotometric assay, zebrafish

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6977 Internet of Things Applications on Supply Chain Management

Authors: Beatriz Cortés, Andrés Boza, David Pérez, Llanos Cuenca

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The Internet of Things (IoT) field is been applied in industries with different purposes. Sensing Enterprise (SE) is an attribute of an enterprise or a network that allows it to react to business stimuli originating on the internet. These fields have come into focus recently on the enterprises and there is some evidence of the use and implications in supply chain management while finding it as an interesting aspect to work on. This paper presents a revision and proposals of IoT applications in supply chain management.

Keywords: industrial, internet of things, production systems, sensing enterprises, sensor, supply chain management

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6976 Distribution of Spotted Fever Group in Ixodid Ticks, Domestic Cattle and Buffalos of Faisalabad District, Punjab, Pakistan

Authors: Muhammad Sohail Sajid, Qurat-ul-Ain, Zafar Iqbal, Muhammad Nisar Khan, Asma Kausar, Adil Ejaz

Abstract:

Rickettsiosis, caused by a Spotted Fever Group Rickettsiae (SFGR), is considered as an emerging infectious disease from public and veterinary perspective. The present study reports distribution of SFGR in the host (buffalo and cattle) and vector (ticks) population determined through gene specific amplification through PCR targeting outer membrane protein (ompA). Tick and blood samples were collected using standard protocols through convenient sampling from district Faisalabad. Ticks were dissected to extract salivary glands (SG). Blood and tick SG pools were subjected to DNA extraction and amplification of ompA using PCR. Overall prevalence of SFGR was reported as 21.5% and 33.6 % from blood and ticks, respectively. Hyalomma anatolicum was more prevalent tick associated with SFGR as compared to Rhipicephalus sp. Higher prevalence of SFGR was reported in cattle (25%) population as compared to that of buffalo (17.07%). On seasonal basis, high SFGR prevalence was recorded during spring season (48.1%, 26.32%, 17.76%) as compared to winter (27.9%, 21.43%, 15.38%) in vector and host (cattle and buffalo respectively) population. Sequencing analysis indicated that rickettsial endo-symbionts were associated with ticks of the study area. These results provided baseline information about the prevalence of SFGR in vector and host population.

Keywords: Rickettsia, livestock, polymerase chain reaction, sequencing, ticks, vectors

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6975 In silico Model of Transamination Reaction Mechanism

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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6974 Integrated Lateral Flow Electrochemical Strip for Leptospirosis Diagnosis

Authors: Wanwisa Deenin, Abdulhadee Yakoh, Chahya Kreangkaiwal, Orawon Chailapakul, Kanitha Patarakul, Sudkate Chaiyo

Abstract:

LipL32 is an outer membrane protein present only on pathogenic Leptospira species, which are the causative agent of leptospirosis. Leptospirosis symptoms are often misdiagnosed with other febrile illnesses as the clinical manifestations are non-specific. Therefore, an accurate diagnostic tool for leptospirosis is indeed critical for proper and prompt treatment. Typical diagnosis via serological assays is generally performed to assess the antibodies produced against Leptospira. However, their delayed antibody response and complicated procedure are undoubtedly limited the practical utilization especially in primary care setting. Here, we demonstrate for the first time an early-stage detection of LipL32 by an integrated lateral-flow immunoassay with electrochemical readout (eLFIA). A ferrocene trace tag was monitored via differential pulse voltammetry operated on a smartphone-based device, thus allowing for on-field testing. Superior performance in terms of the lowest detectable limit of detection (LOD) of 8.53 pg/mL and broad linear dynamic range (5 orders of magnitude) among other sensors available thus far was established. Additionally, the developed test strip provided a straightforward yet sensitive approach for diagnosis of leptospirosis using the collected human sera from patients, in which the results were comparable to the real-time polymerase chain reaction technique.

Keywords: leptospirosis, electrochemical detection, lateral flow immunosensor, point-of-care testing, early-stage detection

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6973 Reaction Kinetics of Biodiesel Production from Refined Cottonseed Oil Using Calcium Oxide

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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6972 Competition Between the Effects of Pesticides and Immune-activation on the Expression of Toll Pathway Genes

Authors: Dani Sukkar, Ali Kanso, Philippe Laval-Gilly, Jairo Falla-Angel

Abstract:

The honeybees' immune system is challenged by different risk factors that induce various responses. However, complex scenarios where bees are exposed to different pesticides simultaneously with immune activation are not well evaluated. The Toll pathway is one of the main signaling pathways studied in invertebrate immune responses, and it is a good indicator of the effect of such complex interactions in addition to key signaling elements of other pathways like Relish of the immune deficiency (IMD) pathway or Eater, the phagocytosis receptor or vitellogenin levels. Honeybee hemocytes extracted from 5th instar larvae were exposed to imidacloprid and/or amitraz with or without the presence of the zymosan a as an immune activator. The gene expression of multiple immune related genes were studied, including spaetzle, Toll, myD88, relish, eater and vitellogenin, by real-time polymerase chain reaction after RNA extraction. The results demonstrated that the Toll pathway is mainly affected by the pesticides; imidacloprid and amitraz, especially by their different combinations. Furthermore, immune activation by zymosan A, a fungal cell-wall component, acts to mitigate to some extent the effect of pesticides on the different levels of the Toll pathway. In addition, imidacloprid, amitraz, and zymosan A have complex and context-specific interactions depending on the levels of immune activation and the pathway evaluated affecting immune-gene expression differently.

Keywords: toll pathway, immune modulation, β-glucan, imidacloprid, amitraz, honeybees, immune genes

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6971 A Study on the Relationship between Transaction Fairness, Social Capital, Supply Chain Integration and Sustainability: Focusing on Manufacturing Companies of South Korea

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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6970 Identification and Characterisation of Oil Sludge Degrading Bacteria Isolated from Compost

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

Abstract:

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

Procedia PDF Downloads 222
6969 Multi-Period Supply Chain Design under Uncertainty

Authors: Amir Azaron

Abstract:

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

Procedia PDF Downloads 275
6968 Plasmodium falciparum and Scistosoma haematobium Co-infection in School Aged Children in Jinduut, Shendam Local Government Area of Plateau State, North Central Nigeria

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

Abstract:

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

Procedia PDF Downloads 303
6967 Theoretical Study of Acetylation of P-Methylaniline Catalyzed by Cu²⁺ Ions

Authors: Silvana Caglieri

Abstract:

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

Procedia PDF Downloads 253
6966 Synthesis and Characterization of Zeolite/Fe3O4 Nanocomposite Material and Investigation of Its Catalytic Reaction

Authors: Mojgan Zendehdel, Safura Molla Mohammad Zamani

Abstract:

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

Procedia PDF Downloads 429
6965 Reverse Logistics, Green Supply Chain, and Carbon Trading

Authors: Neha Asthana, Vishal Krishna Prasad

Abstract:

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

Procedia PDF Downloads 388
6964 Poly(Lactic Acid) Based Flexible Films

Authors: Fathilahbinti Ali, Jamarosliza Jamaluddin, Arun Kumar Upadhyay

Abstract:

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

Procedia PDF Downloads 317
6963 An Efficient and Green Procedure for the Synthesis of Highly Substituted Polyhydronaphthalene Derivatives via a One-Pot, Multi-Component Reaction in Aqueous Media

Authors: Adeleh Moshtaghi Zonouz, Issa Eskandari

Abstract:

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

Procedia PDF Downloads 347