Search results for: metallurgical industrial activity
Commenced in January 2007
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Edition: International
Paper Count: 9156

Search results for: metallurgical industrial activity

7266 The Features of Formation of Russian Agriculture’s Sectoral Structure

Authors: Natalya G. Filimonova, Mariya G. Ozerova, Irina N. Ermakova

Abstract:

The long-term strategy of the economic development of Russia up to 2030 is based on the concept of sustainable growth. The determining factor of such development is complex changes in the economic system which may be achieved by making progressive changes in its structure. The structural changes determine the character and the direction of economic development, as well as they include all elements of this system without exception, and their regulated character ensures the most rapid aim achievement. This article has discussed the industrial structure of the agriculture in Russia. With the use of the system of indexes, the article has determined the directions, intensity, and speed of structural shifts. The influence of structural changes on agricultural production development has been found out. It is noticed that the changes in the industrial structure are synchronized with the changes in the organisation and economic structure. Efficiency assessment of structural changes allowed to trace the efficiency of structural changes and elaborate the main directions for agricultural policy improvement.

Keywords: Russian agricultural sectors, sectoral structure, organizational and economic structure, structural changes

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7265 Sequential Pulsed Electric Field and Ultrasound Assisted Extraction of Bioactive Enriched Fractions from Button Mushroom Stalks

Authors: Bibha Kumari, Nigel P. Brunton, Dilip K. Rai, Brijesh K. Tiwari

Abstract:

Edible mushrooms possess numerous functional components like homo- and hetero- β-glucans [β(1→3), β(1→4) and β(1→6) glucosidic linkages], chitins, ergosterols, bioactive polysaccharides and peptides imparting health beneficial properties to mushrooms. Some of the proven biological activities of mushroom extracts are antioxidant, antimicrobial, immunomodulatory, cholesterol lowering activity by inhibiting a key cholesterol metabolism enzyme i.e. 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCR), angiotensin I-converting enzyme (ACE) inhibition. Application of novel extraction technologies like pulsed electric field (PEF) and high power ultrasound offers clean, green, faster and efficient extraction alternatives with enhanced and good quality extracts. Sequential PEF followed by ultrasound assisted extraction (UAE) were applied to recover bioactive enriched fractions from industrial white button mushroom (Agaricus bisporus) stalk waste using environmentally friendly and GRAS solvents i.e. water and water/ethanol combinations. The PEF treatment was carried out at 60% output voltage, 2 Hz frequency for 500 pulses of 20 microseconds pulse width, using KCl salt solution of 0.6 mS/cm conductivity by the placing 35g of chopped fresh mushroom stalks and 25g of salt solution in the 4x4x4cm3 treatment chamber. Sequential UAE was carried out on the PEF pre-treated samples using ultrasonic-water-bath (USB) of three frequencies (25 KHz, 35 KHz and 45 KHz) for various treatment times (15-120 min) at 80°C. Individual treatment using either PEF or UAE were also investigation to compare the effect of each treatment along with the combined effect on the recovery and bioactivity of the crude extracts. The freeze dried mushroom stalk powder was characterised for proximate compositional parameters (dry weight basis) showing 64.11% total carbohydrate, 19.12% total protein, 7.21% total fat, 31.2% total dietary fiber, 7.9% chitin (as glucosamine equivalent) and 1.02% β-glucan content. The total phenolic contents (TPC) were determined by the Folin-Ciocalteu procedure and expressed as gallic-acid-equivalents (GAE). The antioxidant properties were ascertained using DPPH and FRAP assays and expressed as trolox-equivalents (TE). HMGCR activity and molecular mass of β-glucans will be measured using the commercial HMG-CoA Reductase Assay kit (Sigma-Aldrich) and size exclusion chromatography (HPLC-SEC), respectively. Effects of PEF, UAE and their combination on the antioxidant capacity, HMGCR inhibition and β-glucans content will be presented.

Keywords: β-glucan, mushroom stalks, pulsed electric field (PEF), ultrasound assisted extraction (UAE)

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7264 Framework to Organize Community-Led Project-Based Learning at a Massive Scale of 900 Indian Villages

Authors: Ayesha Selwyn, Annapoorni Chandrashekar, Kumar Ashwarya, Nishant Baghel

Abstract:

Project-based learning (PBL) activities are typically implemented in technology-enabled schools by highly trained teachers. In rural India, students have limited access to technology and quality education. Implementing typical PBL activities is challenging. This study details how Pratham Education Foundation’s Hybrid Learning model was used to implement two PBL activities related to music in 900 remote Indian villages with 46,000 students aged 10-14. The activities were completed by 69% of groups that submitted a total of 15,000 videos (completed projects). Pratham’s H-Learning model reaches 100,000 students aged 3-14 in 900 Indian villages. The community-driven model engages students in 20,000 self-organized groups outside of school. The students are guided by 6,000 youth volunteers and 100 facilitators. The students partake in learning activities across subjects with the support of community stakeholders and offline digital content on shared Android tablets. A training and implementation toolkit for PBL activities is designed by subject experts. This toolkit is essential in ensuring efficient implementation of activities as facilitators aren’t highly skilled and have limited access to training resources. The toolkit details the activity at three levels of student engagement - enrollment, participation, and completion. The subject experts train project leaders and facilitators who train youth volunteers. Volunteers need to be trained on how to execute the activity and guide students. The training is focused on building the volunteers’ capacity to enable students to solve problems, rather than developing the volunteers’ subject-related knowledge. This structure ensures that continuous intervention of subject matter experts isn’t required, and the onus of judging creativity skills is put on community members. 46,000 students in the H-Learning program were engaged in two PBL activities related to Music from April-June 2019. For one activity, students had to conduct a “musical survey” in their village by designing a survey and shooting and editing a video. This activity aimed to develop students’ information retrieval, data gathering, teamwork, communication, project management, and creativity skills. It also aimed to identify talent and document local folk music. The second activity, “Pratham Idol”, was a singing competition. Students participated in performing, producing, and editing videos. This activity aimed to develop students’ teamwork and creative skills and give students a creative outlet. Students showcased their completed projects at village fairs wherein a panel of community members evaluated the videos. The shortlisted videos from all villages were further evaluated by experts who identified students and adults to participate in advanced music workshops. The H-Learning framework enables students in low resource settings to engage in PBL and develop relevant skills by leveraging community support and using video creation as a tool. In rural India, students do not have access to high-quality education or infrastructure. Therefore designing activities that can be implemented by community members after limited training is essential. The subject experts have minimal intervention once the activity is initiated, which significantly reduces the cost of implementation and allows the activity to be implemented at a massive scale.

Keywords: community supported learning, project-based learning, self-organized learning, education technology

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7263 Hepatotoxicity Induced by Arsenic Trioxide in Adult Mice and Their Progeny

Authors: Bouaziz H., Soudania N., Essafia M., Ben Amara I., Hakim A., Jamoussi K., Zeghal Km, Zeghal N.

Abstract:

In this investigation, we have evaluated the effects of arsenic trioxide on hepatic function in pregnant and lactating Swiss albino mice and their suckling pups. Experiments were carried out on female mice given 175 ppm As2O3 in their drinking water from the 14th day of pregnancy until day 14 after delivery. Our results showed a significant decrease in plasma levels of total protein and albumin, cholesterol and triglyceride in As2O3 treated mice and their pups. The hyperbilirubinemia and the increased plasma total alkaline phosphatase activity suggested the presence of cholestasis. Transaminase activities as well as lactate deshydrogenase activity in plasma, known as biomarkers of hepatocellular injury, were elevated indicating hepatic cells’damage after treatment with As2O3. Exposure to arsenic led to an increase of liver thiobarbituric acid reactive substances level along with a concomitant decrease in the activities of superoxide dismutase, catalase and glutathione peroxidase and in glutathione.

Keywords: antioxidant status, arsenic trioxide, hepatotoxicity, mice, oxidative stress

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7262 Analyzing the Relationship between the Spatial Characteristics of Cultural Structure, Activities, and the Tourism Demand

Authors: Deniz Karagöz

Abstract:

This study is attempt to comprehend the relationship between the spatial characteristics of cultural structure, activities and the tourism demand in Turkey. The analysis divided into four parts. The first part consisted of a cultural structure and cultural activity (CSCA) index provided by principal component analysis. The analysis determined four distinct dimensions, namely, cultural activity/structure, accessing culture, consumption, and cultural management. The exploratory spatial data analysis employed to determine the spatial models of cultural structure and cultural activities in 81 provinces in Turkey. Global Moran I indices is used to ascertain the cultural activities and the structural clusters. Finally, the relationship between the cultural activities/cultural structure and tourism demand was analyzed. The raw/original data of the study official databases. The data on the cultural structure and activities gathered from the Turkish Statistical Institute and the data related to the tourism demand was provided by the Republic of Turkey Ministry of Culture and Tourism.

Keywords: cultural activities, cultural structure, spatial characteristics, tourism demand, Turkey

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7261 Study of the Antimicrobial Activity of Aminoreductone against Pathogenic Bacteria in Comparison with Other Antibiotics

Authors: Vu Thu Trang, Lam Xuan Thanh, Samira Sarter, Tomoko Shimamura, Hiroaki Takeuchi  

Abstract:

Antimicrobial activities of aminoreductone (AR), a product formed in the initial stage of Maillard reaction, were screened against pathogenic bacteria. A significant growth inhibition of AR against all 7 isolates (Staphylococcus aureus ATCC® 25923™, Salmonella Typhimurium ATCC® 14028™, Bacillus cereus ATCC® 13061™, Bacillus subtilis ATCC® 11774™, Escherichia coli ATCC® 25922™, Enterococcus faecalis ATCC® 29212™, Listeria innocua ATCC® 33090™) were observed by the standard disc diffusion methods. The inhibition zone for each isolate by AR (2.5 mg) ranged from 15±0 mm to 28.3±0.4 mm in diameter. The minimum inhibitory concentration (MIC) of AR ranging from 20 mM to 26 mM was proven in the seven isolates tested. AR also showed the similar effect of growth inhibition in comparison with antibiotics frequently used for the treatment of infections bacteria, such as amikacin, ciprofloxacin, meropennem, and levofloxacin. The results indicated that foods containing AR are valuable sources of bioactive compounds towards pathogenic bacteria.

Keywords: pathogenic bacteria, aminoreductone, Maillard reaction, antimicrobial activity

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7260 Research on the Function Optimization of China-Hungary Economic and Trade Cooperation Zone

Authors: Wenjuan Lu

Abstract:

China and Hungary have risen from a friendly and comprehensive cooperative relationship to a comprehensive strategic partnership in recent years, and the economic and trade relations between the two countries have developed smoothly. As an important country along the ‘Belt and Road’, Hungary and China have strong economic complementarities and have unique advantages in carrying China's industrial transfer and economic transformation and development. The construction of the China-Hungary Economic and Trade Cooperation Zone, which was initiated by the ‘Sino-Hungarian Borsod Industrial Zone’ and the ‘Hungarian Central European Trade and Logistics Cooperation Park’ has promoted infrastructure construction, optimized production capacity, promoted industrial restructuring, and formed brand and agglomeration effects. Enhancing the influence of Chinese companies in the European market has also promoted economic development in Hungary and even in Central and Eastern Europe. However, as the China-Hungary Economic and Trade Cooperation Zone is still in its infancy, there are still shortcomings such as small scale, single function, and no prominent platform. In the future, based on the needs of China's cooperation with ‘17+1’ and China-Hungary cooperation, on the basis of appropriately expanding the scale of economic and trade cooperation zones and appropriately increasing the number of economic and trade cooperation zones, it is better to focus on optimizing and adjusting its functions and highlighting different economic and trade cooperation. The differentiated function of the trade zones strengthens the multi-faceted cooperation of economic and trade cooperation zones and highlights its role as a platform for cooperation in information, capital, and services.

Keywords: ‘One Belt, One Road’ Initiative, China-Hungary economic and trade cooperation zone, function optimization, Central and Eastern Europe

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7259 Pyrethroid Resistance and Its Mechanism in Field Populations of the Sand Termite, Psammotermes hypostoma Desneux

Authors: Mai. M. Toughan, Ahmed A. A. Sallam, Ashraf O. Abd El-Latif

Abstract:

Termites are eusocial insects that are found on all continents except Antarctica. Termites have serious destructive impact, damaging local huts and crops of poor subsistence. The annual cost of termite damage and its control is determined in the billions globally. In Egypt, most of these damages are due to the subterranean termite species especially the sand termite, P. hypostoma. Pyrethroids became the primary weapon for subterranean termite control, after the use of chlorpyrifos as a soil termiticide was banned. Despite the important role of pyrethroids in termite control, its extensive use in pest control led to the eventual rise of insecticide resistance which may make many of the pyrethroids ineffective. The ability to diagnose the precise mechanism of pyrethroid resistance in any insect species would be the key component of its management at specified location for a specific population. In the present study, detailed toxicological and biochemical studies was conducted on the mechanism of pyrethroid resistance in P. hypostoma. The susceptibility of field populations of P. hypostoma against deltamethrin, α-cypermethrin and ƛ-cyhalothrin was evaluated. The obtained results revealed that the workers of P. hypostoma have developed high resistance level against the tested pyrethroids. Studies carried out through estimation of detoxification enzyme activity indicated that enhanced esterase and cytochrome P450 activities were probably important mechanisms for pyrethroid resistance in field populations. Elevated esterase activity and also additional esterase isozyme were observed in the pyrethroid-resistant populations compared to the susceptible populations. Strong positive correlation between cytochrome P450 activity and pyrethroid resistance was also reported. |Deltamethrin could be recommended as a resistance-breaking pyrethroid that is active against resistant populations of P. hypostoma.

Keywords: Psammotermes hypostoma, pyrethroid resistance, esterase, cytochrome P450

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7258 Photocatalytic Activity of Polypyrrole/ZnO Composites for Degradation of Dye Reactive Red 45 in Wastewater

Authors: Ljerka Kratofil Krehula, Vanja Gilja, Andrea Husak, Sniježana Šuka, Zlata Hrnjak-Murgić

Abstract:

Zinc oxide (ZnO) can be used as photocatalysts for water purification. However, one particular interest is given on the integration of inorganic ZnO nanoclusters with conducting polymers because the resulting nanocomposites may possess unique properties and enhanced photocatalytic activity in comparison to pure ZnO, using UV and also visible light. It is needed to explore the appropriate structure of polypyrrole that can induce activation of ZnO photocatalyst since the synthesis of organic/inorganic hybrid materials can result in a synergistic and complementary feature, increasing ZnO photocatalytic efficiency. In this paper several different composites of polypyrrole/zinc oxide (ZnO) were studied. Composite samples were characterized by X-ray powder diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), cyclic voltammetry (CV) and scanning electron microscopy (SEM). The photocatalytic efficiency of prepared samples was studied as a decomposition of Reactive Red 45 (RR 45) dye, which was monitored by UV-Vis spectroscopy as a change in absorbance of characteristic wavelength at 542 nm. Results show good photocatalytic efficiency of all nanocomposite samples.

Keywords: photocatalysis, polypyrrole, wastewater, zinc oxide

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7257 Through 7S Model to Promote the Service Innovation Management

Authors: Cheng Fang Hsu

Abstract:

Call center is the core of building customer relationship management system. Under the strong competitive stress, it becomes a new profiting challenge for a successful enterprise. Call center is a department not only to provide customer service but also to bring business profit. This is the qualitative case study in Taiwan bank service industry which goes on deeper exploration, and analysis by business interviews and industrial analysis. This study starts from the establishment, development, and management after the reforming of the case call center. Through SWOT analysis, and industrial analysis, this study adopted 7S model to explain how the call center reforms from service oriented to profit oriented and from cost management to profit management. The results indicated how service innovation management promotes call center to be operated as a market profit competition center. The recommendations are indicated to support the call center on marketing profit by service innovation management.

Keywords: call center, 7S model, service innovation management, bioinformatics

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7256 Relevance in the Water-Energy-Food nexus: an Opportunity for Promoting Socio Economic Development in Algeria

Authors: Nadjib Drouiche

Abstract:

Water resources in Algeria are scarce, often low quality, fragile, and unevenly distributed in space and time. The pressure on water resources can be associated with industrial development, a steady population growth, and demanding land irrigation measures. These conditions createa tense competitionfor managing waterresourcesand sharing thembetween agricultural development, drinking water supply, industrial activities, etc. Moreover, the impact of climate change has placed in the forefront national policies focused on the water-energy-food nexus (WEF). In this context, desalination membrane technologies could play an increasing rolefor supporting segments of the Algerian economy that are heavily water-dependent. By implementing water reuse and desalination strategies together in the agricultural sector, there is an opportunity to expand the access to healthy food and clean water, thereby keeping the WEF nexus effects under control.

Keywords: desalination, mitigation, climate change, sustainable development goals

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7255 Phytochemical Investigation, Leaf Structure and Antimicrobial Screening of Pistacia lentiscus against Multi-Drug Resistant Bacteria

Authors: S. Mamoucha, N.Tsafantakis, T. Ioannidis, S. Chatzipanagiotou, C. Nikolaou, L. Skaltsounis, N. Fokialakis, N. Christodoulakis

Abstract:

Introduction: Pistacia lentiscus L. (well known as Mastic tree) is an evergreen sclerophyllous shrub that extensively thrives in the eastern Mediterranean area yet only the trees cultivated in the southern region of the Greek island Chios produces mastic resin. Different parts of P. lentiscus L. var. chia have been used in folk medicine for various purposes, such as tonic, aphrodisiac, antiseptic, antihypertensive and management of dental, gastrointestinal, liver, urinary, and respiratory tract disorders. Several studies have focused on the antibacterial activity of its resin (gum) and its essential oil. However, there is no study combining anatomy of the plant organs, phytochemical profile, and antibacterial screening of the plant. In our attempt to discover novel bioactive metabolites from the mastic tree, we screened its antibacterial activity not only against ATCC strains but also against clinical, resistant strains. Materials-methods: Leaves were investigated using Transmission (ΤΕΜ) and Scanning Εlectron Microscopy (SEM). Histochemical tests were performed on fresh and fixed tissue. Extracts prepared from dried, powdered leaves using 3 different solvents (DCM, MeOH and H2O) the waste water obtained after a hydrodistillation process for essential oil production were screened for their phytochemical content and antibacterial activity. Μetabolite profiling of polar and non-polar extracts was recorded by GC-MS and LC-HRMS techniques and analyzed using in-house and commercial libraries. The antibacterial screening was performed against Staphylococcus aureus ATCC25923, Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853 and against clinical, resistant strains Methicillin-resistant S. aureus (MRSA), Carbapenem-Resistant Metallo-β-Lactamase (carbapenemase) P. aeruginosa (VIM), Klebsiella pneumoniae carbapenemases (KPCs) and Acinetobacter baumanii resistant strains. The antibacterial activity was tested by the Kirby Bauer and the Agar Well Diffusion method. The zone of inhibition (ZI) of each extract was measured and compared with those of common antibiotics. Results: Leaf is compact with inosclereids and numerous idioblasts containing a globular, spiny crystal. The major nerves of the leaf contain a resin duct. Mesophyll cells showed accumulation of osmiophillic metabolites. Histochemical treatments defined secondary metabolites in subcellular localization. The phytochemical investigation revealed the presence of a large number of secondary metabolites, belonging to different chemical groups, such as terpenoids, phenolic compounds (mainly myricetin, kaempferol and quercetin glycosides), phenolic, and fatty acids. Among the extracts, the hydrostillation wastewater achieved the best results against most of the bacteria tested. MRSA, VIM and A. baumanii were inhibited. Conclusion: Extracts from plants have recently been of great interest with respect to their antimicrobial activity. Their use emerged from a growing tendency to replace synthetic antimicrobial agents with natural ones. Leaves of P. lentiscus L. var. chia showed a high antimicrobial activity even against drug - resistant bacteria. Future prospects concern the better understanding of mode of action of the antibacterial activity, the isolation of the most bioactive constituents and the clarification if the activity is related to a single compound or to the synergistic effect of several ones.

Keywords: antibacterial screening, leaf anatomy, phytochemical profile, Pistacia lentiscus var. chia

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7254 Calibration of 2D and 3D Optical Measuring Instruments in Industrial Environments at Submillimeter Range

Authors: Alberto Mínguez-Martínez, Jesús de Vicente y Oliva

Abstract:

Modern manufacturing processes have led to the miniaturization of systems and, as a result, parts at the micro-and nanoscale are produced. This trend seems to become increasingly important in the near future. Besides, as a requirement of Industry 4.0, the digitalization of the models of production and processes makes it very important to ensure that the dimensions of newly manufactured parts meet the specifications of the models. Therefore, it is possible to reduce the scrap and the cost of non-conformities, ensuring the stability of the production at the same time. To ensure the quality of manufactured parts, it becomes necessary to carry out traceable measurements at scales lower than one millimeter. Providing adequate traceability to the SI unit of length (the meter) to 2D and 3D measurements at this scale is a problem that does not have a unique solution in industrial environments. Researchers in the field of dimensional metrology all around the world are working on this issue. A solution for industrial environments, even if it is not complete, will enable working with some traceability. At this point, we believe that the study of the surfaces could provide us with a first approximation to a solution. Among the different options proposed in the literature, the areal topography methods may be the most relevant because they could be compared to those measurements performed using Coordinate Measuring Machines (CMM’s). These measuring methods give (x, y, z) coordinates for each point, expressing it in two different ways, either expressing the z coordinate as a function of x, denoting it as z(x), for each Y-axis coordinate, or as a function of the x and y coordinates, denoting it as z (x, y). Between others, optical measuring instruments, mainly microscopes, are extensively used to carry out measurements at scales lower than one millimeter because it is a non-destructive measuring method. In this paper, the authors propose a calibration procedure for the scales of optical measuring instruments, particularizing for a confocal microscope, using material standards easy to find and calibrate in metrology and quality laboratories in industrial environments. Confocal microscopes are measuring instruments capable of filtering the out-of-focus reflected light so that when it reaches the detector, it is possible to take pictures of the part of the surface that is focused. Varying and taking pictures at different Z levels of the focus, a specialized software interpolates between the different planes, and it could reconstruct the surface geometry into a 3D model. As it is easy to deduce, it is necessary to give traceability to each axis. As a complementary result, the roughness Ra parameter will be traced to the reference. Although the solution is designed for a confocal microscope, it may be used for the calibration of other optical measuring instruments by applying minor changes.

Keywords: industrial environment, confocal microscope, optical measuring instrument, traceability

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7253 The Role of Neuroserpin in Rheumatoid Arthritis Patients

Authors: Sevil Arabaci Tamer, Gonul Gurol, Ibrahim Tekeoglu, Halil Harman, Ihsan Hakki Ciftci

Abstract:

Neuroserpin (NSP) is a serine protease inhibitor and member of the serpin family. It is expressed in developing and adult nervous systems, and acts as an inhibitor of protease tissue plasminogen activator (tPA) and a regulator of neuronal growth and plasticity. Also NSP displays anti-inflammatory activity. But, its role in rheumatoid arthritis had never been studied before. So, the aim of the present study was to investigate the effect of neuroserpin in patients with RA. A total of 50 frozen (-20 ºC) serum samples 40 of them belonged to patients with RA, and 10 sample belonged to healthy subjects, were enrolled prospectively. We used DAS-28 to evaluate disease activity. The following clinical data gathered from the original patients' charts. Serum neuroserpin levels were measured by enzyme-linked immunosorbent assay. Our preliminary study results demonstrate, for the first time, that NSP levels are significantly different in RA patients relative to healthy subjects (P = 0.014). So, NSP contribute to pathological condition of RA. Thus, we believe that serum NSP levels can be as a marker in patients with RA. However other inflammatory diseases should be further investigated.

Keywords: neuroserpin, rheumatoid arthritis, tPA, tPA inhibitor

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7252 Comparative Study on the Effect of Substitution of Li and Mg Instead of Ca on Structural and Biological Behaviors of Silicate Bioactive Glass

Authors: Alireza Arab, Morteza Elsa, Amirhossein Moghanian

Abstract:

In this study, experiments were carried out to achieve a promising multifunctional and modified silicate based bioactive glass (BG). The main aim of the study was investigating the effect of lithium (Li) and magnesium (Mg) substitution, on in vitro bioactivity of substituted-58S BG. Moreover, it is noteworthy to state that modified BGs were synthesized in 60SiO2–(36-x)CaO–4P2O5–(x)Li2O and 60SiO2–(36-x)CaO–4P2O5–(x)MgO (where x = 0, 5, 10 mol.%) quaternary systems, by sol-gel method. Their performance was investigated through different aspects such as biocompatibility, antibacterial activity as well as their effect on alkaline phosphatase (ALP) activity, and proliferation of MC3T3 cells. The antibacterial efficiency was evaluated against methicillin-resistant Staphylococcus aureus bacteria. To do so, CaO was substituted with Li2O and MgO up to 10 mol % in 58S-BGs and then samples were immersed in simulated body fluid up to 14 days and then, characterized by X-ray diffraction, Fourier transform infrared spectroscopy, inductively coupled plasma atomic emission spectrometry, and scanning electron microscopy. Results indicated that this modification led to a retarding effect on in vitro hydroxyapatite (HA) formation due to the lower supersaturation degree for nucleation of HA compared with 58s-BG. Meanwhile, magnesium revealed further pronounced effect. The 3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) and ALP analysis illustrated that substitutions of both Li2O and MgO, up to 5 mol %, had increasing effect on biocompatibility and stimulating proliferation of the pre-osteoblast MC3T3 cells in comparison to the control specimen. Regarding to bactericidal efficiency, the substitution of either Li or Mg for Ca in the 58s BG composition led to statistically significant difference in antibacterial behaviors of substituted-BGs. Meanwhile, the sample containing 5 mol % CaO/Li2O substitution (BG-5L) was selected as a multifunctional biomaterial in bone repair/regeneration due to the improved biocompatibility, enhanced ALP activity and antibacterial efficiency among all of the synthesized L-BGs and M-BGs.

Keywords: alkaline, alkaline earth, bioactivity, biomedical applications, sol-gel processes

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7251 Characterization of Brewery Wastewater Composition

Authors: Abimbola M. Enitan, Josiah Adeyemo, Sheena Kumari, Feroz M. Swalaha, Faizal Bux

Abstract:

With the competing demand on water resources and water reuse, discharge of industrial effluents into the aquatic environment has become an important issue. Much attention has been placed on the impact of industrial wastewater on water bodies worldwide due to the accumulation of organic and inorganic matter in the receiving water bodies. The scope of the present work is to assess the physic-chemical composition of the wastewater produced from one of the brewery industry in South Africa. This is to estimate the environmental impact of its discharge into the receiving water bodies or the municipal treatment plant. The parameters monitored for the quantitative analysis of brewery wastewater include biological oxygen demand (BOD5), chemical oxygen demand (COD), total suspended solids, volatile suspended solids, ammonia, total oxidized nitrogen, nitrate, nitrite, phosphorus, and alkalinity content. In average, the COD concentration of the brewery effluent was 5340.97 mg/l with average pH values of 4.0 to 6.7. The BOD and the solids content of the wastewater from the brewery industry were high. This means that the effluent is very rich in organic content and its discharge into the water bodies or the municipal treatment plant could cause environmental pollution or damage the treatment plant. In addition, there were variations in the wastewater composition throughout the monitoring period. This might be as a result of different activities that take place during the production process, as well as the effects of the peak period of beer production on the water usage.

Keywords: Brewery wastewater, environmental pollution, industrial effluents, physic-chemical composition

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7250 Development of Composite Material for Thermal and Electrical Insulation

Authors: Elmo Thiago Lins Cöuras Ford, Valentina Alessandra Carvalho do Vale, Rubens Maribondo do Nascimento, José Ubiragi de Lima Mendes

Abstract:

Recycling has been greatly stimulated by the market. There are already several products that are produced with recycled materials and various wastes have been studied in various forms of applications. The vast majority of insulation applications in domestic, commercial and industrial systems in the range of low and medium temperatures (up to 180 ° C), using the aggressive nature materials such as glass wool, rock wool, polyurethane, polystyrene. Such materials, while retaining the effectiveness of the heat flux, are disposed as expensive and take years too absorbed by nature. Thus, trying to adapt to a global policy on the preservation of the environment, a study in order to develop an insulating compound of natural / industrial waste and biodegradable materials conducted. Thus, this research presents the development of a composite material based zest tire and latex for thermal and electrical insulation.

Keywords: composite, latex, scrapes tire, insulation, electrical

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7249 Mn3O4-NiFe Layered Double Hydroxides(LDH)/Carbon Composite Cathode for Rechargeable Zinc-Air Battery

Authors: L. K. Nivedha, V. Maruthapandian, R. Kothandaraman

Abstract:

Rechargeable zinc-air batteries (ZAB) are gaining significant research attention owing to their high energy density and copious zinc resources worldwide. However, the unsolved obstacles such as dendrites, passivation, depth of discharge and the lack of an efficient cathode catalyst restrict their practical application1. By and large, non-noble transition metal-based catalysts are well-reputed materials for catalysing oxygen reduction reaction (ORR) and oxygen evolution reaction (OER) with greater stability in alkaline medium2. Herein, we report the synthesis and application of Mn₃O4-NiFeLDH/Carbon composite as a cathode catalyst for rechargeable ZAB. The synergetic effects of the mixed transition metals (Mn/Ni/Fe) have aided in catalysing ORR and OER in alkaline electrolyte with a shallow potential gap of 0.7 V. The composite, by its distinctive physicochemical characteristics, shows an excellent OER activity with a current density of 1.5 mA cm⁻² at a potential of 1.6 V and a superior ORR activity with an onset potential of 0.8 V when compared with their counterparts. Nevertheless, the catalyst prefers a two-electron pathway for the electrochemical reduction of oxygen which results in a limiting current density of 2.5 mA cm⁻². The bifunctional activity of the Mn₃O₄-NiFeLDH/Carbon composite was utilized in developing rechargeable ZAB. The fully fabricated ZAB delivers an open circuit voltage of 1.4 V, a peak power density of 70 mW cm⁻², and a specific capacity of 800 mAh g⁻¹ at a current density of 20 mA cm⁻² with an average discharge voltage of 1 V and the cell is operable upto 50 mA cm-2. Rechargeable ZAB demonstrated over 110 h at 10 mA cm⁻². Further, the cause for the diminished charge-discharge performance experienced beyond the 100th cycle was investigated, and carbon corrosion was testified using Infrared spectroscopy.

Keywords: rechargeable zinc-air battery, oxygen evolution reaction, bifunctional catalyst, alkaline medium

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7248 Preformed Au Colloidal Nanoparticles Immobilised on NiO as Highly Efficient Heterogeneous Catalysts for Reduction of 4-Nitrophenol to 4-Aminophenol

Authors: Khaled Alshammari

Abstract:

A facile approach to synthesizing highly active and stable Au/NiO catalysts for the hydrogenation of nitro-aromatics is reported. Preformed gold nanoparticles have been immobilized onto NiO using a colloidal method. In this article, the reduction of 4-nitrophenol with NaBH4 has been used as a model reaction to investigate the catalytic activity of synthesized Au/NiO catalysts. In addition, we report a systematic study of the reduction kinetics and the influence of specific reaction parameters such as (i) temperature, (ii) stirring rate, (iii) sodium borohydride concentration and (iv) substrate/metal molar ratio. The reaction has been performed at a substrate/metal molar ratio of 7.4, a ratio significantly higher than previously reported. The reusability of the catalyst has been examined, with little to no decrease in activity observed over 5 catalytic cycles. Systematic variation of Au loading reveals the successful synthesis of low-cost and efficient Au/NiO catalysts at very low Au content and using high substrate/metal molar ratios.

Keywords: nonochemistry, catalyst, nanoparticles supported, characterization of materials, colloidal nanoparticles

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7247 Biochemical and Pomological Variability among 14 Moroccan and Foreign Cultivars of Prunus dulcis

Authors: H. Hanine, H. H'ssaini, M. Ibno Alaoui, A. Nablousi, H. Zahir, S. Ennahli, H. Latrache, H. Zine Abidine

Abstract:

Biochemical and pomological variability among 14 cultivars of Prunus dulcis planted in a germoplasm collection site in Morocco were evaluated. Almond samples from six local and eight foreign cultivars (France, Italy, Spain, and USA) were characterized. Biochemical and pomological data revealed significant genetic variability among the 14 cultivars; local cultivars exhibited higher total polyphenol content. Oil content ranged from 35 to 57% among cultivars; both Texas and Toundout genotypes recorded the highest oil content. Total protein concentration from select cultivars ranged from 50 mg/g in Ferraduel to 105 mg/g in Rizlane1 cultivars. Antioxidant activity of almond samples was examined by a DPPH (1,1-diphenyl-2-picrylhydrazyl) radical-scavenging assay; the antioxidant activity varied significantly within the cultivars, with IC50 (the half-maximal inhibitory concentration) values ranging from 2.25 to 20 mg/ml. Autochthonous cultivars originated from the Oujda region exhibited higher tegument total polyphenol and amino acid content compared to others. The genotype Rizlane2 recorded the highest flavonoid content. Pomological traits revealed a large variability within the almond germplasms. The hierarchical clustering analysis of all the data regarding pomological traits distinguished two groups with some particular genotypes as distinct cultivars, and groups of cultivars as polyclone varieties. These results strongly exhibit a potential use of Moroccan-originated almonds as potential clones for future selection due to their nutritional values and pomological traits compared to well-established cultivars.

Keywords: antioxidant activity, DDPH, Moroccan almonds, Prunus dulcis

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7246 The Effect of Calcining Temperature on Photocatalytic Activity of Porous ZnO Architecture

Authors: M. Masar, P. Janota, J. Sedlak, M. Machovsky, I. Kuritka

Abstract:

Zinc oxide (ZnO) nano crystals assembled porous architecture was prepared by thermal decomposition of zinc oxalate precursor at various temperatures ranging from 400-900°C. The effect of calcining temperature on structure and morphology was examined by scanning electron microscopy (SEM), X-ray diffractometry, thermogravimetry, and BET adsorption analysis. The porous nano crystalline ZnO morphology was developed due to the release of volatile precursor products, while the overall shape of ZnO micro crystals was retained as a legacy of the precursor. The average crystallite size increased with increasing temperature of calcination from approximately 21 nm to 79 nm, while the specific surface area decreased from 30 to 1.7 m2g-1. The photo catalytic performance of prepared ZnO powders was evaluated by degradation of methyl violet 2B, a model compound. The significantly highest photo catalytic activity was achieved with powder calcined at 500°C. This may be attributed to the sufficiently well-developed crystalline arrangement, while the specific surface area is still high enough.

Keywords: ZnO, porous structure, photodegradation, methyl violet

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7245 Screening of New Antimicrobial Agents from Heterocyclic Derivatives

Authors: W. Mazari, K. Boucherit, Z. Boucherit-Otmani, M. N. Rahmoun, M. Benabdallah

Abstract:

The hospital or any other establishment of care can be considered as an ecosystem where the patient comes into contact with a frightening microbial universe and a risk to contract infection that is referred to as nosocomial or health care-associated. In these last years, the incidence of these infections has risen sharply. Several microorganisms are the cause of these nosocomial infections and the emergence of resistance of the microbial strains against antibiotics creates a danger to public health. The search for new antimicrobial agents to overcome this problem has produced interesting compounds through chemical synthesis, which plays a very important role in the research and discovery of new drugs. It is in this framework that our study was conducted at our laboratory and it involves evaluating the antibacterial activity of thirteen 2-pyridone derivatives synthesized by two methods, the diffusion disc method and the dilution method against eight Gram negative bacterial strains. The results seem interesting especially for two products that have shown the best activities against Escherichia coli ATCC 25922 and Enterobacter cloacae ATCC 13047 with CMI of 512µg/ml.

Keywords: heterocyclic derivatives, chemical synthesis, antimicrobial activity, biotechnology

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7244 Protective Effect of Rosemary Extract against Toxicity Induced by Egyptian Naja haje Venom

Authors: Walaa H. Salama, Azza M. Abdel-Aty, Afaf S. Fahmy

Abstract:

Background: Egyptian Cobra; Naja haje (Elapidae) is one of most common snakes, widely distributed in Egypt and its envenomation causes multi-organ failure leading to rapid death. Thus, Different medicinal plants showed a protective effect against venom toxicity and may complement the conventional antivenom therapy. Aim: The present study was designed to assess both the antioxidant capacity of methanolic extract of rosemary leaves and evaluate the neutralizing ability of the extract against hepatotoxicity induced by Naja haje venom. Methods: The total phenolic and flavonoid contents and the antioxidant capacity of the methanolic rosemary extract were estimated by DPPH and ABTS Scavenging methods. In addition, the rosemary extract were assessed for anti-venom properties under in vitro and in vivo standard assays. Results: The rosemary extract had high total phenolic and flavonoid content as 12 ± 2 g of gallic acid equivalent per 100 gram of dry weight (g GAE/100g dw) and 5.5 ± 0.8 g of catechin equivalent per 100 grams of dry weight (g CE/100g dw), respectively. In addition, the rosemary extract showed high antioxidant capacity. Furthermore, The rosemary extract were inhibited in vitro the enzymatic activities of phospholipase A₂, L-amino acid oxidase, and hyaluronidase of the venom in a dose-dependent manner. Moreover, indirect hemolytic activity, hepatotoxicity induced by venom were completely neutralized as shown by histological studies. Conclusion: The phenolic compounds of rosemary extract with potential antioxidant activity may be considered as a promising candidate for future therapeutics in snakebite therapy.

Keywords: antioxidant activity, neutralization, phospholipase A₂ enzyme, snake venom

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7243 Effect of Water Addition on Catalytic Activity for CO2 Purification from Oxyfuel Combustion

Authors: Joudia Akil, Stephane Siffert, Laurence Pirault-Roy, Renaud Cousin, Christophe Poupin

Abstract:

Oxyfuel combustion is a promising method that enables to obtain a CO2 rich stream, with water vapor ( ̴10%), unburned components such as CO and NO, which must be cleaned before the use of CO2. Our objective is then the final treatment of CO and NO by catalysis. Three-way catalysts are well-developed material for simultaneous conversion of NO, CO and hydrocarbons. Pt and/or Rh ensure a quasi-complete removal of NOx, CO and HC and there is also a growing interest in partly replacing Pt with less-expensive Pd. The use of alumina and ceria as support ensures, respectively, the stabilization of such species in active state and discharging or storing oxygen to control the oxidation of CO and HC and the reduction of NOx. In this work, we will compare different metals (Pd, Rh and Pt) supported on Al2O3 and CeO2, for CO2 purification from oxyfuel combustion. The catalyst must reduce NO by CO in an oxidizing environment, in the presence of CO2 rich stream and resistant to water. In this study, Al2O3 and CeO2 were used as support materials of the catalysts. 1wt% M/Support where M = Pd, Rh or Pt catalysts were obtained by wet impregnation on supports with a precursor of palladium [Pd(acac)2], rhodium [Rh(NO3)3] and platinum [Pt(NO2)2(NO3)2]. Materials were characterized by BET surface area, H2 chemisorption, and TEM. Catalytic activity was evaluated in CO2 purification which is carried out in a fixed-bed flow reactor containing 150 mg of catalyst at atmospheric pressure. The flow of the reactant gases is composed of: 20% CO2, 10% O2, 0.5% CO, 0.02% NO and 8.2% H2O (He as eluent gas) with a total flow of 200 mL.min−1, with same GHSV (2.24x104 h-1). The catalytic performances of the samples were investigated with and without water. It shows that the total oxidation of CO occurred over the different materials. This study evidenced an important effect of the nature of the metals, supports and the presence or absence of H2O during the reduction of NO by CO in oxyfuel combustions conditions. Rh based catalysts show that the addition of water has a very positive influence especially on the Rh catalyst on CeO2. Pt based catalysts keep a good activity despite the addition of water on the both supports studied. For the NO reduction, addition of water act as a poison with Pd catalysts. The interesting results of Rh based catalysts with water can be explained by a production of hydrogen through the water gas shift reaction. The produced hydrogen acts as a more effective reductant than CO for NO removal. Furthermore, in TWCs, Rh is the main component responsible for NOx reduction due to its especially high activity for NO dissociation. Moreover, cerium oxide is a promotor for WGSR.

Keywords: carbon dioxide, environmental chemistry, heterogeneous catalysis

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7242 Sonochemically Prepared Non-Noble Metal Oxide Catalysts for Methane Catalytic Combustion

Authors: Przemyslaw J. Jodlowski, Roman J. Jedrzejczyk, Damian K. Chlebda, Anna Dziedzicka, Lukasz Kuterasinski, Anna Gancarczyk, Maciej Sitarz

Abstract:

The aim of this study was to obtain highly active catalysts based on non-noble metal oxides supported on zirconia prepared via a sonochemical method. In this study, the influence of the stabilizers addition during the preparation step was checked. The final catalysts were characterized by using such characterization methods as X-ray Diffraction (XRD), nitrogen adsorption, X-ray fluorescence (XRF), scanning electron microscopy (SEM) equipped with energy dispersive X-ray spectrometer (EDS), transmission electron microscopy (TEM) and µRaman. The proposed preparation method allowed to obtain uniformly dispersed metal-oxide nanoparticles at the support’s surface. The catalytic activity of prepared catalyst samples was measured in a methane combustion reaction. The activity of the catalysts prepared by the sonochemical method was considerably higher than their counterparts prepared by the incipient wetness method.

Keywords: methane catalytic combustion, nanoparticles, non-noble metals, sonochemistry

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7241 Photocatalytic Degradation of Methylene Blue Dye Using Cuprous Oxide/Graphene Nanocomposite

Authors: Bekan Bogale, Tsegaye Girma Asere, Tilahun Yai, Fekadu Melak

Abstract:

Aims: To study photocatalytic degradation of methylene blue dye on cuprous oxide/graphene nanocomposite. Background: Cuprous oxide (Cu2O) nanoparticles are among the metal oxides that demonstrated photocatalytic activity. However, the stability of Cu2O nanoparticles due to the fast recombination rate of electron/hole pairs remains a significant challenge in their photocatalytic applications. This, in turn, leads to mismatching of the effective bandgap separation, tending to reduce the photocatalytic activity of the desired organic waste (MB). To overcome these limitations, graphene has been combined with cuprous oxides, resulting in cuprous oxide/graphene nanocomposite as a promising photocatalyst. Objective: In this study, Cu2O/graphene nanocomposite was synthesized and evaluated for its photocatalytic performance of methylene blue (MB) dye degradation. Method: Cu2O/graphene nanocomposites were synthesized from graphite powder and copper nitrate using the facile sol-gel method. Batch experiments have been conducted to assess the applications of the nanocomposites for MB degradation. Parameters such as contact time, catalyst dosage, and pH of the solution were optimized for maximum MB degradation. The prepared nanocomposites were characterized by using UV-Vis, FTIR, XRD, and SEM. The photocatalytic performance of Cu2O/graphene nanocomposites was compared against Cu2O nanoparticles for cationic MB dye degradation. Results: Cu2O/graphene nanocomposite exhibits higher photocatalytic activity for MB degradation (with a degradation efficiency of 94%) than pure Cu2O nanoparticles (67%). This has been accomplished after 180 min of irradiation under visible light. The kinetics of MB degradation by Cu2O/graphene composites can be demonstrated by the second-order kinetic model. The synthesized nanocomposite can be used for more than three cycles of photocatalytic MB degradation. Conclusion: This work indicated new insights into Cu2O/graphene nanocomposite as high-performance in photocatalysis to degrade MB, playing a great role in environmental protection in relation to MB dye.

Keywords: methylene blue, photocatalysis, cuprous oxide, graphene nanocomposite

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7240 Inhibitory Mechanism of Ag and Fe Colloidal Nanoparticles on P. aeruginosa and E.coli Growth

Authors: Fatemeh Moradian, Razieh Ghorbani, Poria Biparva

Abstract:

Growing resistance of microorganisms to potent antibiotics has renewed a great interest towards investigating bactericidal properties of nanoparticles and their Nano composites as an alternative. The use of metal nanoparticles to combat bacterial infections is one of the most wide spread applications of nanotechnology in the field of antibacterial. Nanomaterials have unique properties compared to their bulk counterparts. In this report, we demonstrate the antimicrobial activity of zerovalent Iron(ZVI) and Ag(silver) nanoparticles against Gram-negative bacteria E.coli(DH5α) and Pseudomonas aeruginosa. At first ZVI and Ag nanoparticles were synthesized by chemical reduction method and using scanning electron microscopy (SEM) the nanoparticle size determined. Different concentrations of Ag and ZVI nanoparticles were added to bacteria on nutrient agar medium. Minimum inhibitory concentration (MIC) of Ag and Fe nanoparticles for P. aeruginosa were 5µM and 1µg as well as for E.coli were 6µM. and 10 µg, respectively. Among the two nanoparticles, ZVI showed that the greatest antimicrobial activity against E.coli and Ag nanoparticle on P.aeruginosa. Results suggested that the bactericidal effect of metal nanoparticles has been attributed to their small size as well as high surface to volume ratio and NPs could be used as an effective antibacterial material.

Keywords: bactericidal properties, MIC, nanoparticle, SEM

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7239 Involvement of BCRP/ABCG2 in Protective Mechanisms of Resveratrol against Methotrexate-Induced Renal Damage in Rats

Authors: Mohamed A. Morsy, Azza A. El-Sheikh, Abdulla Y. Al-Taher

Abstract:

Resveratrol (RES) is a well-known polyphenol antioxidant. We have previously shown that testicular protective effect of RES against the anticancer drug methotrexate (MTX)-induced toxicity involves transporter-mediated mechanisms. Here, we investigated the effect of RES on MTX-induced nephrotoxicity. Rats were administered RES (10 mg/kg/day) for 8 days, with or without a single MTX dose (20 mg/kg i.p.) at day 4 of the experiment. MTX induced nephrotoxicity evident by significantly increase in serum blood urea nitrogen and creatinine compared to control, as well as distortion of kidney microscopic structure. MTX also significantly increased renal nitric oxide level, with induction of inducible nitric oxide synthase expression. MTX also significantly up-regulated fas ligand and caspase 3. Administering RES prior to MTX significantly improved kidney function and microscopic picture, as well as significantly decreased nitrosative and apoptotic markers compared to MTX alone. RES, but not MTX, caused significant increase in expression of breast cancer resistance protein (BCRP), an apical efflux renal transporter that participates in urinary elimination of both MTX and RES. Interestingly, concomitant MTX and RES caused further up-regulation of renal Bcrp compared to RES alone. Using Human BCRP ATPase assay, both RES and MTX exhibited dose-dependent increase in ATPase activity, with Km values of 0.52 ± 0.03 and 30.9 ± 4.2 µM, respectively. Furthermore, combined RES and MTX caused ATPase activity which was significantly less than maximum ATPase activity attained by the positive control; sulfasalazine (12.5 µM). In conclusion, RES exerted nephro-protection against MTX-induced toxicity through anti-nitrosative and anti-apoptotic effects, as well as via up-regulation of renal Bcrp.

Keywords: methotrexate, resveratrol, nephrotoxicity, breast cancer resistance protein

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7238 Environmental Impacts on the British Era Structures of Faisalabad-a Detailed Study of the Clock Tower of Faisalabad

Authors: Bazla Manzoor, Aqsa Yasin

Abstract:

Pakistan is the country which is progressing by leaps and bounds through agricultural and industrial growth. The main area, which presents the largest income rate through industrial activities, is Faisalabad from the Province of Punjab. Faisalabad’s main occupations include agriculture and industry. As these sectors i.e. agriculture and industry is developing day by day, they are earning much income for the country and generating thousands of job vacancies. On one hand the city, i.e. Faisalabad is on the way of development through industrial growth, while on the other hand this industrial growth is producing a bad impact on the environment. In return, that damaged environment is affecting badly on the people and built environment. This research is chiefly based on one of the above-mentioned factors i.e. adverse environmental impacts on the built structures. Faisalabad is an old city, therefore; it is having many old structures especially from British Era. Many of those structures are still surviving and are functioning as the government, private and public buildings. However, these structures are getting in a poor condition with the passage of time due to bad maintenance and adverse environmental impacts. Bad maintenance is a factor, which can be controlled by financial assistance and management. The factor needs to be seriously considered is the other one i.e. adverse environmental impacts on British Era structures of the city because this factor requires controlled and refined human activities and actions. For this reason, a research was required to conserve the British Era structures of Faisalabad so that these structures can function well. The other reason to conserve them is that these structures are historically important and are the heritage of the city. For doing this research, literature has been reviewed which was present in the libraries of the city. Department of Environment, Town Municipal Administration, Faisalabad Development Authority and Lyallpur Heritage Foundation were visited to collect the existing data available. Various British Era structures were also visited to note down the environmental impacts on them. From all the structures “Clock Tower,” was deeply studied as it is one of the oldest and most important heritage structures of the city because the earlier settlements of the city were planned based on its location by The British Government. The architectural and environmental analyses were done for The Clock Tower. This research study found the deterioration factors of the tower according to which suggestions have been made.

Keywords: lyallpur, heritage, architecture, environment

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7237 Direct Assessment of Cellular Immune Responses to Ovalbumin with a Secreted Luciferase Transgenic Reporter Mouse Strain IFNγ-Lucia

Authors: Martyna Chotomska, Aleksandra Studzinska, Marta Lisowska, Justyna Szubert, Aleksandra Tabis, Jacek Bania, Arkadiusz Miazek

Abstract:

Objectives: Assessing antigen-specific T cell responses is of utmost importance for the pre-clinical testing of prototype vaccines against intracellular pathogens and tumor antigens. Mainly two types of in vitro assays are used for this purpose 1) enzyme-linked immunospot (ELISpot) and 2) intracellular cytokine staining (ICS). Both are time-consuming, relatively expensive, and require manual dexterity. Here, we assess if a straightforward detection of luciferase activity in blood samples of transgenic reporter mice expressing a secreted Lucia luciferase under the transcriptional control of IFN-γ promoter parallels the sensitivity of IFNγ ELISpot assay. Methods: IFN-γ-LUCIA mouse strain carrying multiple copies of Lucia luciferase transgene under the transcriptional control of IFNγ minimal promoter were generated by pronuclear injection of linear DNA. The specificity of transgene expression and mobilization was assessed in vitro using transgenic splenocytes exposed to various mitogens. The IFN-γ-LUCIA mice were immunized with 50mg of ovalbumin (OVA) emulsified in incomplete Freund’s adjuvant three times every two weeks by subcutaneous injections. Blood samples were collected before and five days after each immunization. Luciferase activity was assessed in blood serum. Peripheral blood mononuclear cells were separated and assessed for frequencies of OVA-specific IFNγ-secreting T cells. Results: We show that in vitro cultured splenocytes of IFN-γ-LUCIA mice respond by 2 and 3 fold increase in secreted luciferase activity to T cell mitogens concanavalin A and phorbol myristate acetate, respectively but fail to respond to B cell-stimulating E.coli lipopolysaccharide. Immunization of IFN-γ-LUCIA mice with OVA leads to over 4 fold increase in luciferase activity in blood serum five days post-immunization with a barely detectable increase in OVA-specific, IFNγ-secreting T cells by ELISpot. Second and third immunizations, further increase the luciferase activity and coincidently also increase the frequencies of OVA-specific T cells by ELISpot. Conclusions: We conclude that minimally invasive monitoring of luciferase secretions in blood serum of IFN-γ-LUCIA mice constitutes a sensitive method for evaluating primary and memory Th1 responses to protein antigens. As such, this method may complement existing methods for rapid immunogenicity assessment of prototype vaccines.

Keywords: ELISpot, immunogenicity, interferon-gamma, reporter mice, vaccines

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