Search results for: resistance of variety
5418 Influence of Titanium Addition on Wear Properties of AM60 Magnesium Alloy
Authors: H. Zengin, M. E. Turan, Y. Turen, H. Ahlatci, Y. Sun
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This study aimed for improving wear resistance of AM60 magnesium alloy by Ti addition (0, 0.2, 0.5, 1wt%Ti). An electric resistance furnace was used to produce alloys. Pure Mg together with Al, Al-Ti and Al-Mn were melted at 750 0C in a stainless steel crucible under controlled Ar gas atmosphere and then poured into a metal mould preheated at 250 0C. Microstructure characterizations were performed by light optical (LOM) and scanning electron microscope (SEM) after the wear test. Wear rates and friction coefficients were measured with a pin-on-disk type UTS-10 Tribometer test device under a load of 20N. The results showed that Ti addition altered the morphology and the amount of b-Mg17Al12 phase in the microstructure of AM60 alloy. b-Mg17Al12 phases on the grain boundaries were refined with increasing amount of Ti. An improvement in wear resistance of AM60 alloy was observed due to the alteration in the microstructure by Ti addition.Keywords: magnesium alloy, titanium, SEM, wear
Procedia PDF Downloads 3335417 Bacteriological Quality of Commercially Prepared Fermented Ogi (AKAMU) Sold in Some Parts of South Eastern Nigeria
Authors: Alloysius C. Ogodo, Ositadinma C. Ugbogu, Uzochukwu G. Ekeleme
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Food poisoning and infection by bacteria are of public health significance to both developing and developed countries. Samples of ogi (akamu) prepared from white and yellow variety of maize sold in Uturu and Okigwe were analyzed together with the laboratory prepared ogi for microbial quality using the standard microbiological methods. The analyses showed that both white and yellow variety had total bacterial counts (cfu/g) of 4.0 ×107 and 3.9 x 107 for the laboratory prepared ogi while the commercial ogi had 5.2 x 107 and 4.9 x107, 4.9 x107 and 4.5 x107, 5.4 x107 and 5.0 x107 for Eke-Okigwe, Up-gate and Nkwo-Achara market respectively. The Staphylococcal counts ranged from 2.0 x 102 to 5.0 x102 and 1.0 x 102 to 4.0 x102 for the white and yellow variety from the different markets while Staphylococcal growth was not recorded on the laboratory prepared ogi. The laboratory prepared ogi had no Coliform growth while the commercially prepared ogi had counts of 0.5 x103 to 1.6 x 103 for white variety and 0.3 x 103 to 1.1 x103 for yellow variety respectively. The Lactic acid bacterial count of 3.5x106 and 3.0x106 was recorded for the laboratory ogi while the commercially prepared ogi ranged from 3.2x106 to 4.2x106 (white variety) and 3.0 x106 to 3.9 x106 (yellow). The presence of bacteria isolates from the commercial and laboratory fermented ogi showed that Lactobacillus sp, Leuconostoc sp and Citrobacter sp were present in all the samples, Micrococcus sp and Klebsiella sp were isolated from Eke-Okigwe and ABSU-up-gate markets varieties respectively, E. coli and Staphylococcus sp were present in Eke-Okigwe and Nkwo-Achara markets while Salmonella sp were isolated from the three markets. Hence, there are chances of contracting food borne diseases from commercially prepared ogi. Therefore, there is the need for sanitary measures in the production of fermented cereals so as to minimize the rate of food borne pathogens during processing and storage.Keywords: ogi, fermentation, bacterial quality, lactic acid bacteria, maize
Procedia PDF Downloads 4065416 Phytochemistry and Biological Activity of Extracts of the Red Raspberry Rubus rosifolius
Authors: Theresa Campbell, Camille Bowen-Forbes, William Aalbersberg
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Differences in the sensory properties of two subtly distinct varieties of Rubus rosifolius lead to the examination of their anthocyanin, essential oil and polyphenol profiles. In both cases, notable differences were identified. Pelargonidin-3-rhutinoside (17.2 mg/100 g FW) and Cyanidin-3-glucoside (66.2 mg/100g FW) proved to be the dominant anthocyanins in the red and wine red varieties respectively. Linalool and terpineol were the major constituents of the essential oil from the red variety; however, those of the wine red variety are unidentified. In regard to phenolic compounds, caffeic acid and quercetin were in a higher concentration in the red variety (1.85 and 0.73 mg/100g FW respectively, compared to 1.22 and 0.34 mg/100g FW respectively in the wine red fruits); while ellagic acid and ferulic acid were of a higher concentration in the wine red variety (0.92 and 0.84mg/100g FW respectively, compared to 0.15 and 0.48 mg/100g FW respectively in the red variety). The methanol extract of both fruit varieties showed great antioxidant activity. Analysis of the antimicrobial activity of the fruit extracts against the growth of drug resistant pathogens revealed that they are active against methicillin resistant S. aureus (MRSA), rifampicin resistant S. aureus (RRSA), wild-type S. aureus (WTSA) and vancomycin-resistant Enterococcus faecium (VREF). Activity was also reported against several food-borne pathogens including two strains of E. coli, L. monocytogenes and Enterobacter aerogenes. The cytotoxicity of the various extracts was assessed and the essential oil extracts exhibited superior activity. The phenolic composition and biological activity of the fruits indicate that their consumption is beneficial to health and also that their incorporation into functional foods and nutraceuticals should be considered.Keywords: phytochemicals, antimicrobial, cytotoxic, Rubus rosifolius
Procedia PDF Downloads 3955415 The Effect of Durability and Pathogen Strains on the Wheat Induced Resistance against Zymoseptoria tritici as a Response to Paenibacillus sp. Strain B2
Authors: E. Samain, T. Aussenac, D. van Tuinen, S. Selim
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Plant growth promoting rhizobacteria are known as potential biofertilizers and plant resistance inducers. The present work aims to study the durability of the resistance induced as a response to wheat seeds inoculation with PB2 and its influence by Z. tritici strains. The internal and external roots colonization have been determined in vitro, seven days post inoculation, by measuring the colony forming unit (CFU). In planta experimentations were done under controlled conditions included four wheat cultivars with different levels of resistance against Septoria Leaf Blotch (SLB) and four Z. tritici strains with high aggressiveness and resistance levels to fungicides. Plantlets were inoculated with PB2 at sowing and infected with Z. tritici at 3 leaves or tillering growth stages. The infection level with SLB was evaluated at 17 days post inoculation using real-time quantitative polymerase chain reaction (PCR). Results showed that PB2 has a high potential of wheat root external colonization (> 10⁶ CFU/g of root). However, the internal colonization seems to be cultivar dependent. Indeed, PB2 has not been observed as endophytic for one cultivar but has a high level of internal colonization with more than 104 CFU/g of root concerning the three others. Two wheat cultivars (susceptible and moderated resistant) were used to investigate PB2-induced resistance (PB2-IR). After the first infection with Z. tritici, results showed that PB2-IR has conferred a high protection efficiency (40-90%) against SLB in the two tested cultivars. Whereas the PB2-IR was effective against all tested strains with the moderate resistant cultivar, it was higher with the susceptible cultivar (> 64%) but against three of the four tested strains. Concerning the durability of the PB2-IR, after the second infection timing, it has been observed a significant decrease (10-59%) depending strains in the moderate resistant cultivar. Contrarily, the susceptible cultivar showed a stable and high protection level (76-84%) but against three of the four tested strains and interestingly, the strain that overcame PB2-IR was not the same as that of the first infection timing. To conclude, PB2 induces a high and durable resistance against Z. tritici. The PB2-IR is pathogen strain, plant growth stage and genotype dependent. These results may explain the loss of the induced resistance effectiveness under field conditions.Keywords: induced resistance, Paenibacillus sp. strain B2, wheat genotypes, Zymoseptoria tritici
Procedia PDF Downloads 1495414 Maintaining Minority Languages; Evidence from Italy
Authors: Carmela Perta
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Following the example of both International and European legislation, on 15 December 1999 the national law 482/99 Regulations regarding the protection of historic language minorities was approved, providing a national framework for the preservation and renaissance of minority languages «The Italian Republic sustains the language and culture of people speaking Albanian, Catalan, German, Greek, Slovene, Croatian, French, Francoprovençal, Friulan, Ladin, Occitan and Sard». The legislation made it possible to use these languages in education, in public offices, in local government, in the judicial system, in mass media, and allowed for the reinstatement of place and personal names. However, several practical problems have emerged, particularly those concerning the variety that should be used in education, in official documents and in other formal domains, i.e. the local variety, the standard of reference (if there is any), or an over regional koinè. In minority settings, it might seem eminently sensible to use the ready made standard of reference, accepting the Ausbausprache, rather than the language as practice, that is the local variety. However, this process seems to be pointless, as is demonstrated by the results of a fieldwork that was carried out in a small town in the South of Italy where members speak Faetar, the local variety of Francoprovençal. Here the language is largely used by the community members in all domains, moreover a deep sense of loyalty towards the variety they use and a manifested minority identity can be observed analysing the speakers’ attitudes. However, these positive attitudes are towards the vehicle for their distinctive history and culture, and not for an “external” standard, a system which local authorities and planners are trying to introduce in the community. In other words, according to the speakers' reactions, there is little point in struggling to maintain a language, if what is conserved is not the group’s language but another.Keywords: maintenance, minority languages, endangered languages, francoprovençal
Procedia PDF Downloads 4335413 The Effects of Fungicide and Genetics on Fungal Diseases on Wheat in Nebraska With Emphasis on Stem Rust
Authors: Javed Sidiqi, Stephen Baezinger, Stephen Wegulo
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Wheat (Triticum aestivum L.) production continues to be challenged by foliar fungal diseases although significant improvement has been made to manage the diseases through developing resistant varieties and the fungicide use to ensure sufficient wheat is produced to meet the growing population’s need. Significant crop losses have been recorded in the history of grain production and yield losses due to fungal diseases, and the trend continues to threat food security in the world and particularly in the less developed countries. The impact of individual fungal diseases on grain yield has been studied extensively to determine crop losses. However, there is limited research available to find out the combined effects of fungal diseases on grain yield and the ways to effectively manage the diseases. Therefore, the objectives of this research were to study the effect of fungal pathogens on grain yield of pre-released winter wheat genotypes in fungicide treated and untreated plots, and to determine whether S7b gene was present in ‘Gage’ wheat as previously hypothesized. Sixty winter wheat genotypes in fungicide treated and untreated plots were studied across four environments. There was a significant effect of fungicide on grain yield consistently across four environments in three years. Fungicide treated wheat lines demonstrated (4,496 kg/ ha-1) grain yield compared to (3,147 kg/ ha-1) grain yield in untreated wheat lines indicating 43% increased grain yield due to severity of foliar fungal diseases. Furthermore, fungicide application also caused an increase in protein concentration from 153 (g kg-1) to 164 (g kg-1) in treated plots in along with test weight from 73 to 77 (kg hL-1) respectively. Gage wheat variety and ISr7b-Ra were crossed to determine presence of Sr7b in Gage. The F2 and F2:3 segregating families were screened and evaluated for stem rust resistance. The segregation of families fell within 15:1 ratio for two separate resistance genes suggesting that Sr7b segregates independently from an unknown resistance gene in Gage that needs to be characterized for its use in the future wheat breeding program to develop resistant wheat varieties.Keywords: funicide, genetics, foliar diseases, grain
Procedia PDF Downloads 1245412 Structural and Functional Comparison of Untagged and Tagged EmrE Protein
Authors: S. Junaid S. Qazi, Denice C. Bay, Raymond Chew, Raymond J. Turner
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EmrE, a member of the small multidrug resistance protein family in bacteria is considered to be the archetypical member of its family. It confers host resistance to a wide variety of quaternary cation compounds (QCCs) driven by proton motive force. Generally, purification yield is a challenge in all membrane proteins because of the difficulties in their expression, isolation and solubilization. EmrE is extremely hydrophobic which make the purification yield challenging. We have purified EmrE protein using two different approaches: organic solvent membrane extraction and hexahistidine (his6) tagged Ni-affinity chromatographic methods. We have characterized changes present between ligand affinity of untagged and his6-tagged EmrE proteins in similar membrane mimetic environments using biophysical experimental techniques. Purified proteins were solubilized in a buffer containing n-dodecyl-β-D-maltopyranoside (DDM) and the conformations in the proteins were explored in the presence of four QCCs, methyl viologen (MV), ethidium bromide (EB), cetylpyridinium chloride (CTP) and tetraphenyl phosphonium (TPP). SDS-Tricine PAGE and dynamic light scattering (DLS) analysis revealed that the addition of QCCs did not induce higher multimeric forms of either proteins at all QCC:EmrE molar ratios examined under the solubilization conditions applied. QCC binding curves obtained from the Trp fluorescence quenching spectra, gave the values of dissociation constant (Kd) and maximum specific one-site binding (Bmax). Lower Bmax values to QCCs for his6-tagged EmrE shows that the binding sites remained unoccupied. This lower saturation suggests that the his6-tagged versions provide a conformation that prevents saturated binding. Our data demonstrate that tagging an integral membrane protein can significantly influence the protein.Keywords: small multidrug resistance (SMR) protein, EmrE, integral membrane protein folding, quaternary ammonium compounds (QAC), quaternary cation compounds (QCC), nickel affinity chromatography, hexahistidine (His6) tag
Procedia PDF Downloads 3765411 Change Management as a Critical Success Factor In E-Government initiatives
Authors: Mohammed Alassim
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In 2014, a UN survey stated that: "The greatest challenge to the adoption of whole-of government, which fundamentally rests on increased collaboration, is resistance to change among government actors". Change management has experienced both theoretically and practically many transformation over the years. When organizations have to implement radical changes, they have to encounter a plethora of issues which leads to ineffective or inefficient implementation of change in most cases. 70% of change projects fail because of human issues. It has been cited that” most studies still show a 60-70% failure rate for organizational change projects — a statistic that has stayed constant from the 1970’s to the present.”. E-government involves not just technical change but cultural, policy, social and organizational evolution. Managing change and overcoming resistance to change is seen as crucial in the success of E-government projects. Resistance can be from different levels in the organization (top management, middle management or employees at operational levels). There can be many reasons for resistance including fear of change and insecurity, lack of knowledge and absence of commitment from management to implement the change. The purpose of this study is to conduct in-depth research to understand the process of change and to identify the critical factors that have led to resistance from employees at different levels (top management, Middle management and operational employees) during e-government initiatives in the public sector in Saudi Arabia. The study is based on qualitative and empirical research methods conducted in the public sector in the Kingdom of Saudi Arabia. This research will use triangulation in data method (interview, group discussion and document review). This research will contribute significantly to knowledge in this field and will identify the measures that can be taken to reduce resistance to change, Upon analysis recommendations or model will be offered which can enable decision makers in public sector in Saudi Arabia how to plan, implement and evaluate change in e-government initiatives via change management strategy.Keywords: change management, e-government, managing change, resistance to change
Procedia PDF Downloads 3155410 Comparison of Entropy Coefficient and Internal Resistance of Two (Used and Fresh) Cylindrical Commercial Lithium-Ion Battery (NCR18650) with Different Capacities
Authors: Sara Kamalisiahroudi, Zhang Jianbo, Bin Wu, Jun Huang, Laisuo Su
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The temperature rising within a battery cell depends on the level of heat generation, the thermal properties and the heat transfer around the cell. The rising of temperature is a serious problem of Lithium-Ion batteries and the internal resistance of battery is the main reason for this heating up, so the heat generation rate of the batteries is an important investigating factor in battery pack design. The delivered power of a battery is directly related to its capacity, decreases in the battery capacity means the growth of the Solid Electrolyte Interface (SEI) layer which is because of the deposits of lithium from the electrolyte to form SEI layer that increases the internal resistance of the battery. In this study two identical cylindrical Lithium-Ion (NCR18650)batteries from the same company with noticeable different in capacity (a fresh and a used battery) were compared for more focusing on their heat generation parameters (entropy coefficient and internal resistance) according to Brandi model, by utilizing potentiometric method for entropy coefficient and EIS method for internal resistance measurement. The results clarify the effect of capacity difference on cell electrical (R) and thermal (dU/dT) parameters. It can be very noticeable in battery pack design for its Safety.Keywords: heat generation, Solid Electrolyte Interface (SEI), potentiometric method, entropy coefficient
Procedia PDF Downloads 4685409 Development of a Large-Scale Cyclic Shear Testing Machine Under Constant Normal Stiffness
Authors: S. M. Mahdi Niktabara, K. Seshagiri Raob, Amit Kumar Shrivastavac, Jiří Ščučkaa
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The presence of the discontinuity in the form of joints is one of the most significant factors causing instability in the rock mass. On the other hand, dynamic loads, including earthquake and blasting induce cyclic shear loads along the joints in rock masses; therefore, failure of rock mass exacerbates along the joints due to changing shear resistance. Joints are under constant normal load (CNL) and constant normal stiffness (CNS) conditions. Normal stiffness increases on the joints with increasing depth, and it can affect shear resistance. For correct assessment of joint shear resistance under varying normal stiffness and number of cycles, advanced laboratory shear machine is essential for the shear test. Conventional direct shear equipment has limitations such as boundary conditions, working under monotonic movements only, or cyclic shear loads with constant frequency and amplitude of shear loads. Hence, a large-scale servo-controlled direct shear testing machine was designed and fabricated to perform shear test under the both CNL and CNS conditions with varying normal stiffness at different frequencies and amplitudes of shear loads. In this study, laboratory cyclic shear tests were conducted on non-planar joints under varying normal stiffness. In addition, the effects of different frequencies and amplitudes of shear loads were investigated. The test results indicate that shear resistance increases with increasing normal stiffness at the first cycle, but the influence of normal stiffness significantly decreases with an increase in the number of shear cycles. The frequency of shear load influences on shear resistance, i.e. shear resistance increases with increasing frequency. However, at low shear amplitude the number of cycles does not affect shear resistance on the joints, but it decreases with higher amplitude.Keywords: cyclic shear load, frequency of load, amplitude of displacement, normal stiffness
Procedia PDF Downloads 1495408 Detection of Transgenes in Cotton (Gossypium hirsutum L.) by using Biotechnology/Molecular Biological Techniques
Authors: Ahmad Ali Shahid, M Shakil Shaukat
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Agriculture is the backbone of economy of Pakistan and Cotton is the major agricultural export and supreme source of raw fiber for our textile industry. To combat against the developing resistance in the target insects and combating these challenges wholesomely, a novel combination of pyramided/stacked genes was conceptualized and later realized, through the means of biotechnology i.e., transformation of three genes namely, Cry1Ac, Cry2A, and EPSP synthase (glyphosate tolerant) genes in the locally cultivated cotton variety. The progenies of the transformed plants were successfully raised and screened under the tunnel conditions for two generations and the present study focused on the screening of plants which were confirmed for containing all of these three genes and their expressions. Initially, the screening was done through glyphosate spray assay and the plants which were healthy and showed no damage on leaves were selected after 07 days of spray. In the laboratory, the DNA of these plants were isolated and subjected to amplification of the three genes. Thus, seventeen out of twenty were confirmed positive for Cry1Ac gene and ten out of twenty were positive for Cry2A gene and all twenty were positive for presence of EPSP synthase gene. Then, the ten plant samples which were confirmed with presence of all three genes were subjected to expression analysis of these proteins through ELISA. The results showed that eight out of ten plants were actively expressing the three transgenes. Real-time PCR was also done to quantify the expression levels of the EPSP synthase gene. Finally, eight plants were confirmed for the presence and active expression of all three genes in T3 generation of the triple gene transformed cotton. These plants may be subjected to T4 generation to develop a new stable variety in due course of time.Keywords: agriculture, cotton, transformation, cry genes, ELISA, PCR
Procedia PDF Downloads 3945407 Real-time PCR to Determine Resistance Genes in ESBLEscherichia Coli Strains Stored in the Epidemic Diseases Laboratory of the National Institute of Hygiene (INH)
Authors: A. Qasmaoui, F. Ohmani, Z. Zaine, I. El Akrad, J. Hamamouchi, K. Halout, B. Belkadi, R. Charof
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The evolution of antibiotic resistance is a crucial aspect of the problem related to the intensive use of these substances in medicine for humans and animals. The production of ESBL extended spectrum β-lactamase enzymes is the main mechanism of resistance to β-lactam antibiotics in Escherichia coli. The objective of our work is to determine the resistance genes in E. coli strains.ESBL coli stored at the epidemic diseases laboratory of the National Institute of Hygiene. The strains were identified according to the classic bacteriological criteria. An antibiogram was performed on the strains isolated by the Mueller Hinton agar disc diffusion method. The production of ESBL in the strains was detected by the synergy assay technique and confirmed for the presence of the blaCTX-M1, blaCTX-M2, blaTEM, blaSHV, blaOXA-48 genes by gene amplification . Of the 27 observed strains of E.coli, 17 isolated strains present the phenotype of extended-spectrum Beta-lactamase with a percentage of 63%.. All 18 cefotaxime-resistant strains were analyzed for an ESBL phenotype. All strains were positive in the double-disc synergy assay. The fight against the emergence and spread of these multi-resistant antibiotic-resistant strains requires the reasonable use of antibiotics.Keywords: E coli, BLSE, CTX, TEM, SHV, OXA, résistance aux antibiotique
Procedia PDF Downloads 165406 Antimicrobial Resistance Patterns of Campylobacter from Pig and Cattle Carcasses in Poland
Authors: Renata Szewczyk, Beata Lachtara, Kinga Wieczorek, Jacek Osek
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Campylobacter is recognized as the main cause of bacterial gastrointestinal infections in Europe. A main source of the pathogen is poultry and poultry meat; however, other animals like pigs and cattle can also be reservoirs of the bacteria. Human Campylobacter infections are often self-limiting but in some cases, macrolide and fluoroquinolones have to be used. The aim of this study was to determine antimicrobial resistance patterns (AMR) of Campylobacter isolated from pig and cattle carcasses. Between July 2009 and December 2015, 735 swabs from pig (n = 457) and cattle (n = 278) carcasses were collected at Polish slaughterhouses. All samples were tested for the presence of Campylobacter by ISO 10272-1 and confirmed to species level using PCR. The antimicrobial susceptibility of Campylobacter isolates was determined by a microbroth dilution method with six antimicrobials: gentamicin (GEN), streptomycin (STR), erythromycin (ERY), nalidixic acid (NAL), ciprofloxacin (CIP) and tetracycline (TET). It was found that 167 of 735 samples (22.7%) were contaminated with Campylobacter. The vast majority of them were of pig origin (134; 80.2%), whereas for cattle carcasses Campylobacter was less prevalent (33; 19.8%). Among positive samples C. coli was predominant species (123; 73.7%) and it was isolated mainly from pig carcasses. The remaining isolates were identified as C. jejuni (44; 26.3%). Antimicrobial susceptibility indicated that 22 out of 167 Campylobacter (13.2%) were sensitive to all antimicrobials used. Fourteen of them were C. jejuni (63.6%; pig, n = 6; cattle, n = 8) and 8 was C. coli (36.4%; pig, n = 4; cattle, n = 4). Most of the Campylobacter isolates (145; 86.8%) were resistant to one or more antimicrobials (C. coli, n = 115; C. jejuni, n = 30). Comparing the AMR for Campylobacter species it was found that the most common pattern for C. jejuni was CIP-NAL-TET (9; 30.0%), whereas CIP-NAL-STR-TET was predominant among C. coli (47; 40.9%). Multiresistance, defined as resistance to three or more classes of antimicrobials, was found in 57 C. coli strains, mostly obtained from pig (52 isolates). On the other hand, only one C. jejuni strain, isolated from cattle, showed multiresistance with pattern CIP-NAL-STR-TET. Moreover, CIP-NAL-STR-TET was characteristic for most of multiresistant C. coli isolates (47; 82.5%). For the remaining C. coli the resistance patterns were CIP-ERY-NAL-TET (7 strains; 12.3%) and for one strain of each patterns: ERY-STR-TET, CIP-STR-TET, CIP-NAL-GEN-STR-TET. According to the present findings resistance to erythromycin was observed only in 11 C. coli (pig, n = 10; cattle, n = 1). In conclusion, the results of this study showed that pig carcasses may be a serious public health concern because of contamination with C. coli that might features multiresistance to antimicrobials.Keywords: antimicrobial resistance, Campylobacter, carcasses, multi resistance
Procedia PDF Downloads 3305405 Experimental Investigation of Hull Form for Electric Driven Ferry
Authors: Vasilij Djackov, Tomas Zapnickas, Evgenii Iamshchikov, Lukas Norkevicius, Rima Mickeviciene, Larisa Vasiljeva
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In this paper, the resistance and pitching values of the test of an electric ferry are presented. The research was carried out in the open flow channel of Klaipėda University with a multi-axis dynamometer. The received model resistance values were recalculated to the real vessel and the preliminary chosen propulsion unit power was compared. After analyzing the results of the pitching of the model, it was concluded that the shape of the hull needs to be further improved, taking into account the possible uneven weight distribution at the ends of the ferry. Further investigation of the hull of the electric ferry is recommended, including experiments with various water depths and activation of propulsion units.Keywords: electrical ferry, model tests, open flow channel, pitching, resistance
Procedia PDF Downloads 925404 Evaluation of Antimicrobial Efficacy of Nanofluid Containing Carbon Nanotubes Functionalized with Antibiotic on Urinary Tract Infection
Authors: Erfan Rahimi, Hadi Bahari Far, Mojgan Shikhpour
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Background: Urinary tract infection is one of the most common nosocomial infections, especially among women. E. coli is one of the main causes of urinary tract infections and one of the most common antibiotics to fight this bacterium is ampicillin. As conventional antibiotics led to bacterial antibiotic resistance, modification of the pure drugs can address this issue. The aim of this study was to prepare nanofluids containing carbon nanotubes conjugated with ampicillin to improve drug performance and reduce antibiotic resistance. Methods: Multi-walled carbon nanotubes (MWCNTs) were activated with thionyl chloride by reflux system and nanofluids containing antibiotics were prepared by ultrasonic method. The properties of the prepared nano-drug were investigated by general element analysis, infrared spectroscopy, Raman spectroscopy, scanning electron microscopy and transmission electron microscopy. After the treatment of the desired strain with nanofluid, microbial studies were performed to evaluate the antibacterial effects and molecular studies were carried out to measure the expression of the resistance gene AcrAB. Result: We have shown that the antimicrobial effect of ampicillin-functionalized MWCNTs at low concentrations performed better than that of the conventional drug in both resistant and ATCC strains. Also, a decrease in antibiotic resistance of bacteria treated with ampicillin-functionalized MWCNTs compared to the pure drug was observed. Also, ampicillin-functionalized MWCNTs downregulated the expression of AcrAB in treated bacteria. Conclusion: Because carbon nanotubes are capable of destroying the bacterial wall, which provides antibiotic resistance features in bacteria, their usage in the form of nanofluids can make lower dosages (about three times less) than that of the pure drug more effective. Additionally, the expression of the bacterial resistance gene AcrAB decreased, thereby reducing antibiotic resistance and improving drug performance against bacteria.Keywords: urinary tract infection, antibiotic resistance, carbon nanotube, nanofluid
Procedia PDF Downloads 1455403 Prevalence and Genetic Determinant of Drug Resistant Tuberculosis among Patients Completing Intensive Phase of Treatment in a Tertiary Referral Center in Nigeria
Authors: Aminu Bashir Mohammad, Agwu Ezera, Abdulrazaq G. Habib, Garba Iliyasu
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Background: Drug resistance tuberculosis (DR-TB) continues to be a challenge in developing countries with poor resources. Routine screening for primary DR-TB before commencing treatment is not done in public hospitals in Nigeria, even with the large body of evidence that shows a high prevalence of primary DR-TB. Data on drug resistance and its genetic determinant among follow up TB patients is lacking in Nigeria. Hence the aim of this study was to determine the prevalence and genetic determinant of drug resistance among follow up TB patients in a tertiary hospital in Nigeria. Methods: This was a cross-sectional laboratory-based study conducted on 384 sputum samples collected from consented follow-up tuberculosis patients. Standard microbiology methods (Zeil-Nielsen staining and microscopy) and PCR (Line Probe Assay)] were used to analyze the samples collected. Person’s Chi-square was used to analyze the data generated. Results: Out of three hundred and eighty-four (384) sputum samples analyzed for mycobacterium tuberculosis (MTB) and DR-TB twenty-five 25 (6.5%) were found to be AFB positive. These samples were subjected to PCR (Line Probe Assay) out of which 18(72%) tested positive for DR-TB. Mutations conferring resistance to rifampicin (rpo B) and isoniazid (katG, and or inhA) were detected in 12/18(66.7%) and 6/18(33.3%), respectively. Transmission dynamic of DR-TB was not significantly (p>0.05) dependent on demographic characteristics. Conclusion: There is a need to strengthened the laboratory capacity for diagnosis of TB and drug resistance testing and make these services available, affordable, and accessible to the patients who need them.Keywords: drug resistance tuberculosis, genetic determinant, intensive phase, Nigeria
Procedia PDF Downloads 2845402 Marker Assisted Selection of Rice Genotypes for Xa5 and Xa13 Bacterial Leaf Blight Resistance Genes
Authors: P. Sindhumole, K. Soumya, R. Renjimol
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Rice (Oryza sativa L.) is the major staple food crop over the world. It is prone to a number of biotic and abiotic stresses, out of which Bacterial Leaf Blight (BLB), caused by Xanthomonas oryzae pv. oryzae, is the most rampant. Management of this disease through chemicals or any other means is very difficult. The best way to control BLB is by the development of Host Plant Resistance. BLB resistance is not an activity of a single gene but it involves a cluster of more than thirty genes reported. Among these, Xa5 and Xa13 genes are two important ones, which can be diagnosed through marker assisted selection using closely linked molecular markers. During 2014, the first phase of field screening using forty traditional rice genotypes was carried out and twenty resistant symptomless genotypes were identified. Molecular characterisation of these genotypes using RM 122 SSR marker revealed the presence of Xa5 gene in thirteen genotypes. Forty-two traditional rice genotypes were used for the second phase of field screening for BLB resistance. Among these, sixteen resistant genotypes were identified. These genotypes, along with two susceptible check genotypes, were subjected to marker assisted selection for Xa13 gene, using the linked STS marker RG-136. During this process, presence of Xa13 gene could be detected in ten resistant genotypes. In future, these selected genotypes can be directly utilised as donors in Marker assisted breeding programmes for BLB resistance in rice.Keywords: oryza sativa, SSR, STS, marker, disease, breeding
Procedia PDF Downloads 3935401 Resistance Spot Welding of Boron Steel 22MnB5 with Complex Welding Programs
Authors: Szymon Kowieski, Zygmunt Mikno
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The study involved the optimization of process parameters during resistance spot welding of Al-coated martensitic boron steel 22MnB5, applied in hot stamping, performed using a programme with a multiple current impulse mode and a programme with variable pressure force. The aim of this research work was to determine the possibilities of a growth in welded joint strength and to identify the expansion of a welding lobe. The process parameters were adjusted on the basis of welding process simulation and confronted with experimental data. 22MnB5 steel is known for its tendency to obtain high hardness values in weld nuggets, often leading to interfacial failures (observed in the study-related tests). In addition, during resistance spot welding, many production-related factors can affect process stability, e.g. welding lobe narrowing, and lead to the deterioration of quality. Resistance spot welding performed using the above-named welding programme featuring 3 levels of force made it possible to achieve 82% of welding lobe extension. Joints made using the multiple current impulse program, where the total welding time was below 1.4s, revealed a change in a peeling mode (to full plug) and an increase in weld tensile shear strength of 10%.Keywords: 22MnB5, hot stamping, interfacial fracture, resistance spot welding, simulation, single lap joint, welding lobe
Procedia PDF Downloads 3855400 Novel Development on Orthopedic Prosthesis by Nanocrystalline Hydroxyapatite Nanocomposite Coated on 316 L Stainless Steel
Authors: Neriman Ozada, Ebrahim Karamian, Amirsalar Khandan, Sina Ghafoorpoor Yazdi
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Natural hydroxyapatite, NHA, coatings on the surface of 316 L stainless steel implants has been widely employed in order to achieve better osteoconductivity. For coating, the plasma spraying method is generally used because they ensure adhesion between the coating and the 316 L stainless steel (SS) surface. Some compounds such as zircon (ZrSiO4) is employed as an additive in an attempt to improve HA’s mechanical properties such as wear resistance and hardness. In this study wear resistance has been carried out in different chemical compositions of coating. Therefore, nanocomposites based on NHA containing of 0 wt.%, 5 wt.%, 10 wt.%, and 15 wt.% of zircon were used as a coating on the SS implants. The samples consisted of NHA, derived from calf heated at 850 °C for 3 h. The composite mixture was coated on SS by plasma spray method. The results were estimated using the scanning electron microscopy (SEM), X-ray diffraction (XRD) techniques were utilized to characterize the shape and size of NHA powder. Disc wear test and Vickers hardness were utilized to characterize the coated nanocomposite samples. The prepared NHA powder had nano-scale morphological structure with the mean crystallite size of 30-50 nm in diameter. The wear resistance are almost 320, 380, 415, and 395 m/g and hardness are approximately 376, 391, 420, 410 VHN in ceramic composite materials containing ZrSiO4. The results have been shown that the best wear resistance and hardness occurred in the sample coated by NHA/ZrSiO4 containing of 10 wt.% of zircon.Keywords: zircon, 316 L stainless steel, wear resistance, orthopedic applications, plasma spray
Procedia PDF Downloads 4325399 Analysis and Modeling of Graphene-Based Percolative Strain Sensor
Authors: Heming Yao
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Graphene-based percolative strain gauges could find applications in many places such as touch panels, artificial skins or human motion detection because of its advantages over conventional strain gauges such as flexibility and transparency. These strain gauges rely on a novel sensing mechanism that depends on strain-induced morphology changes. Once a compression or tension strain is applied to Graphene-based percolative strain gauges, the overlap area between neighboring flakes becomes smaller or larger, which is reflected by the considerable change of resistance. Tiny strain change on graphene-based percolative strain sensor can act as an important leverage to tremendously increase resistance of strain sensor, which equipped graphene-based percolative strain gauges with higher gauge factor. Despite ongoing research in the underlying sensing mechanism and the limits of sensitivity, neither suitable understanding has been obtained of what intrinsic factors play the key role in adjust gauge factor, nor explanation on how the strain gauge sensitivity can be enhanced, which is undoubtedly considerably meaningful and provides guideline to design novel and easy-produced strain sensor with high gauge factor. We here simulated the strain process by modeling graphene flakes and its percolative networks. We constructed the 3D resistance network by simulating overlapping process of graphene flakes and interconnecting tremendous number of resistance elements which were obtained by fractionizing each piece of graphene. With strain increasing, the overlapping graphenes was dislocated on new stretched simulation graphene flake simulation film and a new simulation resistance network was formed with smaller flake number density. By solving the resistance network, we can get the resistance of simulation film under different strain. Furthermore, by simulation on possible variable parameters, such as out-of-plane resistance, in-plane resistance, flake size, we obtained the changing tendency of gauge factor with all these variable parameters. Compared with the experimental data, we verified the feasibility of our model and analysis. The increase of out-of-plane resistance of graphene flake and the initial resistance of sensor, based on flake network, both improved gauge factor of sensor, while the smaller graphene flake size gave greater gauge factor. This work can not only serve as a guideline to improve the sensitivity and applicability of graphene-based strain sensors in the future, but also provides method to find the limitation of gauge factor for strain sensor based on graphene flake. Besides, our method can be easily transferred to predict gauge factor of strain sensor based on other nano-structured transparent optical conductors, such as nanowire and carbon nanotube, or of their hybrid with graphene flakes.Keywords: graphene, gauge factor, percolative transport, strain sensor
Procedia PDF Downloads 4165398 Regulation on Macrophage and Insulin Resistance after Aerobic Exercise in High-Fat Diet Mice
Authors: Qiaofeng Guo
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Aims: Obesity is often accompanied by insulin resistance (IR) and whole-body inflammation. Aerobic exercise is an effective treatment to improve insulin resistance and inflammation. However, the anti-inflammatory mechanisms of exercise on epididymal and subcutaneous adipose remain to be elucidated. Here, we compared the macrophage polarization between epididymal and subcutaneous adipose after aerobic exercise. Methods: Male C57BL/6 mice were fed a normal diet group or a high-fat diet group for 12 weeks and performed aerobic training on a treadmill at 55%~65% VO₂ max for eight weeks. Food intake, body weight, and fasting blood glucose levels were monitored weekly. The intraperitoneal glucose tolerance test was to evaluate the insulin resistance model. Fat mass, blood lipid profile, serum IL-1β, TNF-α levels, and CD31/CD206 rates were analysed after the intervention. Results: FBG (P<0.01), AUCIPGTT (P<0.01), and HOMA-IR (P<0.01) increased significantly for a high-fat diet and decreased significantly after the exercise. Eight weeks of aerobic exercise attenuated HFD-induced weight gain and glucose intolerance and improved insulin sensitivity. Serum IL-1β, TNF-α, CD11C/CD206 expression in subcutaneous adipose tissue were not changed before and after exercise, but not in epididymal adipose tissue (P<0.01). Conclusion: Insulin resistance is not accompanied by chronic inflammation and M1 polarization of subcutaneous adipose tissue macrophages in high-fat diet mice. Aerobic exercise effectively improved lipid metabolism and insulin sensitivity, which may be closely associated with reduced M1 polarization of epididymal adipose macrophages.Keywords: aerobic exercise, insulin resistance, chronic inflammation, adipose, macrophage polarization
Procedia PDF Downloads 765397 Study of Temperature Difference and Current Distribution in Parallel-Connected Cells at Low Temperature
Authors: Sara Kamalisiahroudi, Jun Huang, Zhe Li, Jianbo Zhang
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Two types of commercial cylindrical lithium ion batteries (Panasonic 3.4 Ah NCR-18650B and Samsung 2.9 Ah INR-18650), were investigated experimentally. The capacities of these samples were individually measured using constant current-constant voltage (CC-CV) method at different ambient temperatures (-10 ℃, 0 ℃, 25 ℃). Their internal resistance was determined by electrochemical impedance spectroscopy (EIS) and pulse discharge methods. The cells with different configurations of parallel connection NCR-NCR, INR-INR and NCR-INR were charged/discharged at the aforementioned ambient temperatures. The results showed that the difference of internal resistance between cells much more evident at low temperatures. Furthermore, the parallel connection of NCR-NCR exhibits the most uniform temperature distribution in cells at -10 ℃, this feature is quite favorable for the safety of the battery pack.Keywords: batteries in parallel connection, internal resistance, low temperature, temperature difference, current distribution
Procedia PDF Downloads 4775396 Stenotrophomonas maltophilia: The Major Carbapenem Resistance Bacteria from Waste Water Treatment Plant of Pig Farm
Authors: Young-Ji Kim, Jin-Hyeong Park, Hong-Seok Kim, Jung-Whan Chon, Kwang-Yeop Kim, Dong-Hyeon Kim, Il-Byeong Kang, Da-Na Jeong, Jin-Hyeok Yim, Ho-Seok Jang, Kwang-Young Song, Kun-Ho Seo
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Stenotrophomonas maltophilia is one of the emerging opportunistic pathogens, and also known to have extensive drug resistance intrinsically including carbepenems which is last resort for most serious infections. One possible way for S. maltophilia to infect human is via wastewater treatment plant (WWTP). In the period between October 2016 and February 2017, effluent samples of WWTP from 3 different pig farms were collected once a month and screened for isolation of S. maltophilia. Total 16 strains of S. maltophilia were isolated and, the antibiotic susceptibility phenotypes were determined by Vitek 2 system for 16 antibiotics, ampicillin (AMP), amoxicillin/clavulanic acid (AMC), piperacillin/tazobactam (TZP), cefazolin (CZ), cefoxitin (FOX), cefotaxime (CTX), ceftazidime (CAZ), cefepime (FEP), aztreonam (AZT), ertapenem (ETP), imipenem (IMP), amikacin (AK), gentamicin (GN), ciprofloxacin (CIP), tigecycline (TGC) and trimethoprim/sulfamethoxazole (SXT). All isolates showed high resistance to AMP (100%), CZ (100%), FOX (100%), CTX (100%), CAZ (100%), FEP (94%), AZT (100%), ETP (100%), IMP (100%), AK (100%), GN (100%) whereas were susceptible to CIP (0%), TGC (0%), SXT (6%). All strains harbored at least one of the antibiotic resistance determinant such as spgM, rmlA, and rpfF. Some isolates had similar MLST (multilocus sequence typing) types with clinical isolates, suggesting WWTP could have potential role in the transmission of S. maltophilia to aquatic environment and, possibly, to humans.Keywords: Stenotrophomonas maltophilia, Carbapenem resistance, waste water treatment plant, pig farm
Procedia PDF Downloads 4615395 Wear Resistance of Graphene Oxide and Carbon Nanotubes Silanized Coatings
Authors: Henrique Gomes dos Santosa, Manoel Henrique Alvesa, Jane Zoppas Ferreiraa, Annelise Kopp Alvesa
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This work aimed to seek an environmentally sustainable surface coating alternative by researching the influence of the addition of graphene oxide (GO) and carbon nanotubes (CNT) on the silanization of coatings to increase the wear resistance in galvanized steel, using the pin-on-disk test. The results obtained were compared between different concentrations of additives and the number of coating layers, in addition to comparing with samples without coating and only with silane layers. Bis-1,2-(triethoxysilyl)ethane (BTSE) silane was used in silanizing the coatings with CNT or GO and applied to the samples through dip-coating to form one, four, or eight layers. The wear test results found that three samples stood out in relation to the objective, showing an increase in wear resistance compared to the galvanized sample only. The rolling effect and the lubricity character presented by carbon nanotubes were positive for the increase in wear resistance obtained. The reduction in wear compared to the galvanized-only sample reached 82%. Raman spectroscopy was also carried out to detect the presence of silane, GO, and CNT, in addition to roughness tests and SEM to assess the homogeneity of the coating. The carbonaceous additives, graphene oxide, and carbon nanotubes in certain amounts of layers and specific concentrations fulfilled their objective against the wear imposed on the substrate.Keywords: silane, coating, graphene oxide, carbon nanotubes, wear resistance
Procedia PDF Downloads 115394 Comparison Between Vegans and Omnivores on the Recovery of Delayed Onset Muscle Soreness in Young Females
Authors: Njeim Pressila, Hajj-Boutros Guy, Antony D. Karelis
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Background: Acute resistance exercise is associated with an elevated inflammation response, which could lead to delayed onset muscle soreness (DOMS). There is evidence that suggests that ingesting foods that have anti-inflammation properties may help reduce DOMS. A vegan diet has also been shown to be an anti-inflammatory diet which could, in turn, decrease DOMS. Objective and hypothesis: The purpose of the present study will be to compare markers of DOMS between vegans and omnivores after acute resistance exercise in young females. We hypothesize that vegans will have a better recovery of DOMS markers after a resistance exercise session compared to omnivores. Methods: Population: We will recruit30 vegans and 30 omnivores to participate in this study. Allvolunteers will follow either a vegan or an omnivore diet for at least 2 years. Anthropometric measurements, body composition, musclestrength (leg and chest press), markers of DOMS (swelling, pain, and stiffness), and dietary factors, as well as a wellness and anxiety questionnaire will be measured. All participants will also perform an acute resistance exercise session in order to induce DOMS. Pertinence: This project will give us a better understanding on the recovery process of vegans after a resistance training session and, as such, provide useful information to health professionals and athletes/coaches (kinesiologists and nutritionists)Keywords: vgeans, omnivores, delayed onset muscle soreness, pain, stifness
Procedia PDF Downloads 1265393 Tribocorrosion Behavior of Austempered Ductile Iron Microalloyed with Boron
Authors: S. Gvazava, N. Khidasheli, G. Gordeziani, A. DL. Batako
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The work presented in this paper studied the tribological characteristics (wear resistance, friction coefficient) of austempered ductile iron (ADI) with different combinations of structural composition (upper bainite, lower bainite, retained austenite) in dry sliding friction. A range of structural states of the metal matrix was obtained by changing the regimes of isothermal quenching of high-strength cast iron. The tribological tests were carried out using two sets of isothermal quenched cast irons. After austenitization at 900°С for 60 minutes, the specimens from the first group were isothermally quenched at the 300°С temperature and the specimens from the second set – at 400°С. The investigations showed that the isothermal quenching increases the friction coefficient of high-strength cast irons. The friction coefficient was found to be in the range from 0.4 to 0.55 for cast irons, depending on the structures of the metal matrix. The quenched cast irons having lower bainite demonstrate higher wear resistance in dry friction conditions. The dependence of wear resistance on the amount of retained austenite in isothermal quenched cast irons has a nonlinear characteristic and reaches its maximum value when the content of retained austenite is about 15-22%. The boron micro-additives allowed to reduce the friction coefficient of ADI and increase their wear resistance by 1.5-1.7 times.Keywords: wear resistance, dry sliding, austempering, ADI, friction coefficient, retained austenite, isothermal quenching
Procedia PDF Downloads 1795392 Using Tyre Ash as Ground Resistance Improvement Material-Health and Environmental Perspective
Authors: George Eduful, Dominic Yeboah, Kingsford Joseph A. Atanga
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The use of tyre ash as backfill material for ground electrode has been found to provide ultra-low and stable ground resistance value for grounding systems. However, health and environmental concerns have been expressed regarding its application. To address these concerns, the paper investigates chemical contents of the tyre ash and compares them to levels considered non-hazardous to health and the environment. It was found that the levels of the pollutant agents in the tyre ash were within the recommended safety margins. The rate of ground electrode corrosion in tyre ash material was also investigated. It was found that the effect of corrosion and the life of electrode can be extended if the tyre ash is mixed with cement. For best results, a ratio of 10 portions of tyre ash to 1 portion of cement is recommended.Keywords: tyre ash, scrapped tyre, ground resistance reducing agent, rate of corrosion
Procedia PDF Downloads 4025391 To Study the Effect of Optic Fibre Laser Cladding of Cast Iron with Silicon Carbide on Wear Rate
Authors: Kshitij Sawke, Pradnyavant Kamble, Shrikant Patil
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The study investigates the effect on wear rate of laser clad of cast iron with silicon carbide. Metal components fail their desired use because they wear, which causes them to lose their functionality. The laser has been used as a heating source to create a melt pool over the surface of cast iron, and then a layer of hard silicon carbide is deposited. Various combinations of power and feed rate of laser have experimented. A suitable range of laser processing parameters was identified. Wear resistance and wear rate properties were evaluated and the result showed that the wear resistance of the laser treated samples was exceptional to that of the untreated samples.Keywords: laser clad, processing parameters, wear rate, wear resistance
Procedia PDF Downloads 2565390 Glutamine Supplementation and Resistance Traning on Anthropometric Indices, Immunoglobulins, and Cortisol Levels
Authors: Alireza Barari, Saeed Shirali, Ahmad Abdi
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Introduction: Exercise has contradictory effects on the immune system. Glutamine supplementation may increase the resistance of the immune system in athletes. The Glutamine is one of the most recognized immune nutrients that as a fuel source, substrate in the synthesis of nucleotides and amino acids and is also known to be part of the antioxidant defense. Several studies have shown that improving glutamine levels in plasma and tissues can have beneficial effects on the function of immune cells such as lymphocytes and neutrophils. This study aimed to investigate the effects of resistance training and training combined with glutamine supplementation to improve the levels of cortisol and immunoglobulin in untrained young men. The research shows that physical training can increase the cytokines in the athlete’s body of course; glutamine can counteract the negative effects of resistance training on immune function and stability of the mast cell membrane. Materials and methods: This semi-experimental study was conducted on 30 male non-athletes. They were randomly divided into three groups: control (no exercise), resistance training, resistance training and glutamine supplementation, respectively. Resistance training for 4 weeks and glutamine supplementation in 0.3 gr/kg/day after practice was applied. The resistance-training program consisted of eight exercises (leg press, lat pull, chest press, squat, seatedrow, abdominal crunch, shoulder press, biceps curl and triceps press down) four times per week. Participants performed 3 sets of 10 repetitions at 60–75% 1-RM. Anthropometry indexes (weight, body mass index, and body fat percentage), oxygen uptake (VO2max) Maximal, cortisol levels of immunoglobulins (IgA, IgG, IgM) were evaluated Pre- and post-test. Results: Results showed four week resistance training with and without glutamine cause significant increase in body weight, BMI and significantly decreased (P < 0/001) in BF. Vo2max also increased in both groups of exercise (P < 0/05) and exercise with glutamine (P < 0/001), such as in both groups significant reduction in IgG (P < 0/05) was observed. But no significant difference observed in levels of cortisol, IgA, IgM in any of the groups. No significant change observed in either parameter in the control group. No significant difference observed between the groups. Discussion: The alterations in the hormonal and immunological parameters can be used in order to assess the effect overload on the body, whether acute or chronically. The plasmatic concentration of glutamine has been associated to the functionality of the immunological system in individuals sub-mitted to intense physical training. resistance training has destructive effects on the immune system and glutamine supplementation cannot neutralize the damaging effects of power exercise on the immune system.Keywords: glutamine, resistance traning, immuglobulins, cortisol
Procedia PDF Downloads 4785389 MicroRNA-1246 Expression Associated with Resistance to Oncogenic BRAF Inhibitors in Mutant BRAF Melanoma Cells
Authors: Jae-Hyeon Kim, Michael Lee
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Intrinsic and acquired resistance limits the therapeutic benefits of oncogenic BRAF inhibitors in melanoma. MicroRNAs (miRNA) regulate the expression of target mRNAs by repressing their translation. Thus, we investigated miRNA expression patterns in melanoma cell lines to identify candidate biomarkers for acquired resistance to BRAF inhibitor. Here, we used Affymetrix miRNA V3.0 microarray profiling platform to compare miRNA expression levels in three cell lines containing BRAF inhibitor-sensitive A375P BRAF V600E cells, their BRAF inhibitor-resistant counterparts (A375P/Mdr), and SK-MEL-2 BRAF-WT cells with intrinsic resistance to BRAF inhibitor. The miRNAs with at least a two-fold change in expression between BRAF inhibitor-sensitive and –resistant cell lines, were identified as differentially expressed. Averaged intensity measurements identified 138 and 217 miRNAs that were differentially expressed by 2 fold or more between: 1) A375P and A375P/Mdr; 2) A375P and SK-MEL-2, respectively. The hierarchical clustering revealed differences in miRNA expression profiles between BRAF inhibitor-sensitive and –resistant cell lines for miRNAs involved in intrinsic and acquired resistance to BRAF inhibitor. In particular, 43 miRNAs were identified whose expression was consistently altered in two BRAF inhibitor-resistant cell lines, regardless of intrinsic and acquired resistance. Twenty five miRNAs were consistently upregulated and 18 downregulated more than 2-fold. Although some discrepancies were detected when miRNA microarray data were compared with qPCR-measured expression levels, qRT-PCR for five miRNAs (miR-3617, miR-92a1, miR-1246, miR-1936-3p, and miR-17-3p) results showed excellent agreement with microarray experiments. To further investigate cellular functions of miRNAs, we examined effects on cell proliferation. Synthetic oligonucleotide miRNA mimics were transfected into three cell lines, and proliferation was quantified using a colorimetric assay. Of the 5 miRNAs tested, only miR-1246 altered cell proliferation of A375P/Mdr cells. The transfection of miR-1246 mimic strongly conferred PLX-4720 resistance to A375P/Mdr cells, implying that miR-1246 upregulation confers acquired resistance to BRAF inhibition. We also found that PLX-4720 caused much greater G2/M arrest in A375P/Mdr cells transfected with miR-1246mimic than that seen in scrambled RNA-transfected cells. Additionally, miR-1246 mimic partially caused a resistance to autophagy induction by PLX-4720. These results indicate that autophagy does play an essential death-promoting role inPLX-4720-induced cell death. Taken together, these results suggest that miRNA expression profiling in melanoma cells can provide valuable information for a network of BRAF inhibitor resistance-associated miRNAs.Keywords: microRNA, BRAF inhibitor, drug resistance, autophagy
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