Search results for: broth micro dilution assay
2567 Burrowing Invertebrates Induce Fragmentation of Mariculture Styrofoam Floats and Formation of Microplastics
Authors: Yifan Zheng, Jinmin Zhu, Jiji Li, Gulling Li, Huahong Shi
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Secondary microplastics originate from the fragmentation of large plastics, and weathering is supposed to be the main cause of fragmentation. In this study, we investigated burrows and burrowing invertebrates on Styrofoam floats from the mariculture areas of China’s coastal waters. Various burrows were found on the submerged surface of Styrofoam floats and could be divided into ‘I’, ‘S’, ‘J’, and ‘Y’ types based on the burrow entrance number and passage curvature. Different invertebrate species, including 5 isopods, 8 clamworms, and 12 crabs, were found inside the burrows. Micro-foams were found in the bodies of these burrowers, with an average abundance of 4.2 ± 0.3 (isopod), 6.9 ± 2.0 (clamworm), and 3.0 ± 0.5 (crab) micro-foams per individual. In the laboratory, we observed the boring process of crabs in abandoned floats. Field and laboratory evidence suggested that these invertebrates bored various burrows. The total volume of crab burrows on a 3-year-used float was estimated to be 2.6 × 10³ cm³, producing 4.1 × 10⁸ microplastics. This study highlights the critical role of bioerosion in destroying man-made substrates and prompting microplastic pollution.Keywords: burrowing invertebrate, mariculture area, styrofoam float, fragmentation, microplastics
Procedia PDF Downloads 952566 Development and Validation of High-Performance Liquid Chromatography Method for the Determination and Pharmacokinetic Study of Linagliptin in Rat Plasma
Authors: Hoda Mahgoub, Abeer Hanafy
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Linagliptin (LNG) belongs to dipeptidyl-peptidase-4 (DPP-4) inhibitor class. DPP-4 inhibitors represent a new therapeutic approach for the treatment of type 2 diabetes in adults. The aim of this work was to develop and validate an accurate and reproducible HPLC method for the determination of LNG with high sensitivity in rat plasma. The method involved separation of both LNG and pindolol (internal standard) at ambient temperature on a Zorbax Eclipse XDB C18 column and a mobile phase composed of 75% methanol: 25% formic acid 0.1% pH 4.1 at a flow rate of 1.0 mL.min-1. UV detection was performed at 254nm. The method was validated in compliance with ICH guidelines and found to be linear in the range of 5–1000ng.mL-1. The limit of quantification (LOQ) was found to be 5ng.mL-1 based on 100µL of plasma. The variations for intra- and inter-assay precision were less than 10%, and the accuracy values were ranged between 93.3% and 102.5%. The extraction recovery (R%) was more than 83%. The method involved a single extraction step of a very small plasma volume (100µL). The assay was successfully applied to an in-vivo pharmacokinetic study of LNG in rats that were administered a single oral dose of 10mg.kg-1 LNG. The maximum concentration (Cmax) was found to be 927.5 ± 23.9ng.mL-1. The area under the plasma concentration-time curve (AUC0-72) was 18285.02 ± 605.76h.ng.mL-1. In conclusion, the good accuracy and low LOQ of the bioanalytical HPLC method were suitable for monitoring the full pharmacokinetic profile of LNG in rats. The main advantages of the method were the sensitivity, small sample volume, single-step extraction procedure and the short time of analysis.Keywords: HPLC, linagliptin, pharmacokinetic study, rat plasma
Procedia PDF Downloads 2352565 Simulation and Characterization of Stretching and Folding in Microchannel Electrokinetic Flows
Authors: Justo Rodriguez, Daming Chen, Amador M. Guzman
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The detection, treatment, and control of rapidly propagating, deadly viruses such as COVID-19, require the development of inexpensive, fast, and accurate devices to address the urgent needs of the population. Microfluidics-based sensors are amongst the different methods and techniques for detection that are easy to use. A micro analyzer is defined as a microfluidics-based sensor, composed of a network of microchannels with varying functions. Given their size, portability, and accuracy, they are proving to be more effective and convenient than other solutions. A micro analyzer based on the concept of “Lab on a Chip” presents advantages concerning other non-micro devices due to its smaller size, and it is having a better ratio between useful area and volume. The integration of multiple processes in a single microdevice reduces both the number of necessary samples and the analysis time, leading the next generation of analyzers for the health-sciences. In some applications, the flow of solution within the microchannels is originated by a pressure gradient, which can produce adverse effects on biological samples. A more efficient and less dangerous way of controlling the flow in a microchannel-based analyzer is applying an electric field to induce the fluid motion and either enhance or suppress the mixing process. Electrokinetic flows are characterized by no less than two non-dimensional parameters: the electric Rayleigh number and its geometrical aspect ratio. In this research, stable and unstable flows have been studied numerically (and when possible, will be experimental) in a T-shaped microchannel. Additionally, unstable electrokinetic flows for Rayleigh numbers higher than critical have been characterized. The flow mixing enhancement was quantified in relation to the stretching and folding that fluid particles undergo when they are subjected to supercritical electrokinetic flows. Computational simulations were carried out using a finite element-based program while working with the flow mixing concepts developed by Gollub and collaborators. Hundreds of seeded massless particles were tracked along the microchannel from the entrance to exit for both stable and unstable flows. After post-processing, their trajectories, the folding and stretching values for the different flows were found. Numerical results show that for supercritical electrokinetic flows, the enhancement effects of the folding and stretching processes become more apparent. Consequently, there is an improvement in the mixing process, ultimately leading to a more homogenous mixture.Keywords: microchannel, stretching and folding, electro kinetic flow mixing, micro-analyzer
Procedia PDF Downloads 1172564 Rhizobium leguminosarum: Selecting Strain and Exploring Delivery Systems for White Clover
Authors: Laura Villamizar, David Wright, Claudia Baena, Marie Foxwell, Maureen O'Callaghan
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Leguminous crops can be self-sufficient for their nitrogen requirements when their roots are nodulated with an effective Rhizobium strain and for this reason seed or soil inoculation is practiced worldwide to ensure nodulation and nitrogen fixation in grain and forage legumes. The most widely used method of applying commercially available inoculants is using peat cultures which are coated onto seeds prior to sowing. In general, rhizobia survive well in peat, but some species die rapidly after inoculation onto seeds. The development of improved formulation methodology is essential to achieve extended persistence of rhizobia on seeds, and improved efficacy. Formulations could be solid or liquid. Most popular solid formulations or delivery systems are: wettable powders (WP), water dispersible granules (WG), and granules (DG). Liquid formulation generally are: suspension concentrates (SC) or emulsifiable concentrates (EC). In New Zealand, R. leguminosarum bv. trifolii strain TA1 has been used as a commercial inoculant for white clover over wide areas for many years. Seeds inoculation is carried out by mixing the seeds with inoculated peat, some adherents and lime, but rhizobial populations on stored seeds decline over several weeks due to a number of factors including desiccation and antibacterial compounds produced by the seeds. In order to develop a more stable and suitable delivery system to incorporate rhizobia in pastures, two strains of R. leguminosarum (TA1 and CC275e) and several formulations and processes were explored (peat granules, self-sticky peat for seed coating, emulsions and a powder containing spray dried microcapsules). Emulsions prepared with fresh broth of strain TA1 were very unstable under storage and after seed inoculation. Formulations where inoculated peat was used as the active ingredient were significantly more stable than those prepared with fresh broth. The strain CC275e was more tolerant to stress conditions generated during formulation and seed storage. Peat granules and peat inoculated seeds using strain CC275e maintained an acceptable loading of 108 CFU/g of granules or 105 CFU/g of seeds respectively, during six months of storage at room temperature. Strain CC275e inoculated on peat was also microencapsulated with a natural biopolymer by spray drying and after optimizing operational conditions, microparticles containing 107 CFU/g and a mean particle size between 10 and 30 micrometers were obtained. Survival of rhizobia during storage of the microcapsules is being assessed. The development of a stable product depends on selecting an active ingredient (microorganism), robust enough to tolerate some adverse conditions generated during formulation, storage, and commercialization and after its use in the field. However, the design and development of an adequate formulation, using compatible ingredients, optimization of the formulation process and selecting the appropriate delivery system, is possibly the best tool to overcome the poor survival of rhizobia and provide farmers with better quality inoculants to use.Keywords: formulation, Rhizobium leguminosarum, storage stability, white clover
Procedia PDF Downloads 1422563 Simulation of Reflection Loss for Carbon and Nickel-Carbon Thin Films
Authors: M. Emami, R. Tarighi, R. Goodarzi
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Maximal radar wave absorbing cannot be achieved by shaping alone. We have to focus on the parameters of absorbing materials such as permittivity, permeability, and thickness so that best absorbing according to our necessity can happen. The real and imaginary parts of the relative complex permittivity (εr' and εr") and permeability (µr' and µr") were obtained by simulation. The microwave absorbing property of carbon and Ni(C) is simulated in this study by MATLAB software; the simulation was in the frequency range between 2 to 12 GHz for carbon black (C), and carbon coated nickel (Ni(C)) with different thicknesses. In fact, we draw reflection loss (RL) for C and Ni-C via frequency. We have compared their absorption for 3-mm thickness and predicted for other thicknesses by using of electromagnetic wave transmission theory. The results showed that reflection loss position changes in low frequency with increasing of thickness. We found out that, in all cases, using nanocomposites as absorbance cannot get better results relative to pure nanoparticles. The frequency where absorption is maximum can determine the best choice between nanocomposites and pure nanoparticles. Also, we could find an optimal thickness for long wavelength absorbing in order to utilize them in protecting shields and covering.Keywords: absorbing, carbon, carbon nickel, frequency, thicknesses
Procedia PDF Downloads 1782562 Comparison of Acetylcholinesterase Reactivators Cytotoxicity with Their Structure
Authors: Lubica Muckova, Petr Jost, Jaroslav Pejchal, Daniel Jun
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The development of acetylcholinesterase reactivators, i.e. antidotes against organophosphorus poisoning, is an important goal of defence research. The aim of this study was to compare cytotoxicity and chemical structure of 5 currently available (pralidoxime, trimedoxime, obidoxime, methoxime, and asoxime) and 4 newly developed compounds (K027, K074, K075, and K203). In oximes, there could be at least four important structural factors affecting their toxicity, including the number of oxime groups in the molecule, the position of oxime group(s) on pyridinium ring, the length of carbon linker, and the substitution by oxygen or insertion of the double bond into the connection chain. The cytotoxicity of tested substances was measured using colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay (MTT assay) in SH-SY5Y cell line. Toxicity was expressed as toxicological index IC₅₀. The tested compounds showed different cytotoxicity ranging from 1.5 to 27 mM. K027 was the least, and methoxime was the most toxic reactivator. The lowest toxicity was found in a monopyridinium reactivator and bispyridinium reactivators with simple 3C carbon linker. Shortening of connection chain length to 1C, incorporation of oxygen moiety into 3C compounds, elongation of carbon linker to 4C and insertion of a double bond into 4C substances increase AChE reactivators' cytotoxicity. Acknowledgements: This work was supported by a long-term organization development plan Medical Aspects of Weapons of Mass Destruction of the Faculty of Military Health Sciences, University of Defence.Keywords: acetylcholinesterase, cytotoxicity, organophosphorus poisoning, reactivators of acetylcholinesterase
Procedia PDF Downloads 2992561 Studies on Microstructure and Mechanical Properties of Simulated Heat Affected Zone in a Micro Alloyed Steel
Authors: Sanjeev Kumar, S. K. Nath
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Proper selection of welding parameters for getting excellent weld is a challenge. HAZ simulation helps in identifying suitable welding parameters like heating rate, cooling rate, peak temperature, and energy input. In this study, the influence of weld thermal cycle of heat affected zone (HAZ) is simulated for Submerged Arc Welding (SAW) using Gleeble ® 3800 thermomechanical simulator. A (Micro-alloyed) MA steel plate of thickness 18 mm having yield strength 450MPa is used for making test specimens. Determination of the mechanical properties of weld simulated specimens including Charpy V-notch toughness and hardness is performed. Peak temperatures of 1300°C, 1150°C, 1000°C, 900°C, 800°C, heat energy input of 22KJ/cm and preheat temperatures of 30°C have been used with Rykalin-3D simulation model. It is found that the impact toughness (75J) is the best for the simulated HAZ specimen at the peak temperature 900ºC. For parent steel, impact toughness value is 26.8J at -50°C in transverse direction.Keywords: HAZ simulation, mechanical properties, peak temperature, ship hull steel, weldability
Procedia PDF Downloads 5572560 Sharia, Legal Pluralism and Muslim Personal Law in Contemporary India
Authors: K. C. Mujeebu Rahman
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Over the years, discussions in India regarding personal law in India have focused on its deficiencies, increasing involvement of the judiciary, and the pursuit of uniformity. However, little attention has been given to understanding how the law functions in a multicultural nation committed to political secularism. This paper addresses this gap by exploring the mahallu system in Malabar, shedding light on the decision-making process within Muslim personal law. It reveals that this process is deeply rooted in everyday micro-politics, sectarian dynamics, social pressure, and emotions. Through an in-depth examination of a triple talaq case, the paper demonstrates how love (or the lack of it), family expectations, and community authority intersect in resolving marital disputes. Instead of a straightforward legal interpretation, this process leads to a complex maze of micro-politics involving local religious factions and authorities. The paper underscores that the non-state quasi-legal institutions within the mahallu system represent a distinct form of legal pluralism characterized by intricate power dynamics at multiple levels. Moreover, it highlights the interplay between what is considered legally valid and what is deemed socially legitimate.Keywords: islamic law, sharia, fatwa, muslim personal law
Procedia PDF Downloads 582559 Experimental and Analytical Dose Assessment of Patient's Family Members Treated with I-131
Authors: Marzieh Ebrahimi, Vahid Changizi, Mohammad Reza Kardan, Seyed Mahdi Hosseini Pooya, Parham Geramifar
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Radiation exposure to the patient's family members is one of the major concerns during thyroid cancer radionuclide therapy. The aim of this study was to measure the total effective dose of the family members by means of thermoluminescence personal dosimeter, and compare with those calculated by analytical methods. Eighty-five adult family members of fifty-one patients volunteered to participate in this research study. Considering the minimum and maximum range of dose rate from 15 µsv/h to 120 µsv/h at patients' release time, the calculated mean and median dose values of family members were 0.45 mSv and 0.28 mSv, respectively. Moreover, almost all family members’ doses were measured to be less than the dose constraint of 5 mSv recommended by Basic Safety Standards. Considering the influence parameters such as patient dose rate and administrated activity, the total effective doses of family members were calculated by TEDE and NRC formulas and compared with those of experimental results. The results indicated that, it is fruitful to use the quantitative calculations for releasing patients treated with I-131 and correct estimation of patients' family doses.Keywords: effective dose, thermoluminescence, I-131, thyroid cancer
Procedia PDF Downloads 3882558 Antibacterial Activity of Methanol Extract of Punica Granatum Linn. (Punnicaceae) Fruit Peel Against Selected Bacterial Species
Authors: Afzan Mahmad, Santibuana Abd Rahman, Gouri Kumar Dash, Mohd. Syafiq Bin Abdullah
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Antibacterial activity of the methanol extract of fruit peel of Punica granatum Linn (Family: Punicaceae) was evaluated against two Gram positive and two Gram negative bacteria. The Gram positive bacteria included Staphylococcus aureus, Streptococcus pneumoniae and the Gram negative organisms included Escherichia coli and Pseudomonas aeruginosa respectively. The culture media used for antibacterial assay was Mueller Hinton agar for the growth of S. aureus, E. coli, and P. aeruginosa. The media used for the growth of S. pneumoniae was Mueller Hinton blood agar. The antibacterial assay was performed through Disc diffusion technique. The methanol extract was tested at three different concentrations (50, 100 and 200 mg/ml). Standard antibiotic discs containing vancomycin (30 μg) for S. pneumoniae, penicillin (10 units) for S. aureus, ceftriaxone (30 μg) for E. coli and ciprofloxacin (5 μg) for P. aeruginosa were used for the activity comparison. The results of the study revealed that the extract possesses antibacterial activity against S. aureus, S. pneumoniae and P. aeruginosa at all tested concentrations. The maximum zone of inhibition of 19 mm of the extract at 200 mg/ml was observed against S. pneumoniae. However, no zone of inhibition was observed against E. coli at the tested concentrations of the extract. Based on the results obtained in this study, it may be concluded that the fruit peel of P. granatum possess broad spectrum of antibacterial activity against a number bacteria.Keywords: Punica granatum Linn., methanol extract, antibacterial, zone of inhibition
Procedia PDF Downloads 3842557 Effects of Macro and Micro Nutrients on Growth and Yield Performances of Tomato (Lycopersicon esculentum MILL.)
Authors: K. M. S. Weerasinghe, A. H. K. Balasooriya, S. L. Ransingha, G. D. Krishantha, R. S. Brhakamanagae, L. C. Wijethilke
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Tomato (Lycopersicon esculentum Mill.) is a major horticultural crop with an estimated global production of over 120 million metric tons and ranks first as a processing crop. The average tomato productivity in Sri Lanka (11 metric tons/ha) is much lower than the world average (24 metric tons/ha).To meet the tomato demand for the increasing population the productivity has to be intensified through the agronomic-techniques. Nutrition is one of the main factors which govern the growth and yield of tomato and the main nutrient source soil affect the plant growth and quality of the produce. Continuous cropping, improper fertilizer usage etc., cause widespread nutrient deficiencies. Therefore synthetic fertilizers and organic manures were introduced to enhance plant growth and maximize the crop yields. In this study, effects of macro and micronutrient supplementations on improvement of growth and yield of tomato were investigated. Selected tomato variety is Maheshi and plants were grown in Regional Agricultural and Research Centre Makadura under the Department of Agriculture recommended (DOA) macro nutrients and various combination of Ontario recommended dosages of secondary and micro fertilizer supplementations. There were six treatments in this experiment and each treatment was replicated in three times and each replicate consisted of six plants. Other than the DOA recommendation, five combinations of Ontario recommended dosage of secondary and micronutrients for tomato were also used as treatments. The treatments were arranged in a Randomized Complete Block Design. All cultural practices were carried out according to the DOA recommendations. The mean data was subjected to the statistical analysis using SAS package and mean separation (Duncan’s Multiple Range test at 5% probability level) procedures. Secondary and micronutrients containing treatments significantly increased most of the growth parameters. Plant height, plant girth, number of leaves, leaf area index etc. Fruits harvested from pots amended with macro, secondary and micronutrients performed best in terms of total yield; yield quality; to pots amended with DOA recommended dosage of fertilizer for tomato. It could be due to the application of all essential macro and micro nutrients that rise in photosynthetic activity, efficient translocation and utilization of photosynthates causing rapid cell elongation and cell division in actively growing region of the plant leading to stimulation of growth and yield were caused. The experiment revealed and highlighted the requirements of essential macro, secondary and micro nutrient fertilizer supplementations for tomato farming. The study indicated that, macro and micro nutrient supplementation practices can influence growth and yield performances of tomato fruits and it is a promising approach to get potential tomato yields.Keywords: macro and micronutrients, tomato, SAS package, photosynthates
Procedia PDF Downloads 4552556 Evaluation of Capacity of Bed Planted with Macrophytes for Wastewater Treatment of Biskra City, Algeria
Authors: Mimeche Leila, Debabeche Mahmoud
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It is question to study and to value the possibility of settling the process of purification by plants (constructed wetland) to treat the domestic waste water of Biskra, city in a semi-arid environment with grave problems of. According to the bibliography, the process of treatment by plants is considered as more advantageous than the classic techniques. It is the use of beds with macrophytes where the purification is made by the combined action of plants and micro-organisms in a filtering bed. The micro-organisms which are aerobic bacteria and\or anaerobic have for main function to degrade the polluting materials. Plants in the macrophytes beds have for function to serve as support in the development of bacteria and to favour also their development. In this study, we present a preliminary experimental analysis of the potentialities of treatment of some macrpohytes plants, implanted in basins filled of gravel. Analyses physico chemical and bacteriological of the waste water indicate a good elimination of the polluting materials, and put in evidence the purifier power of these plants, in association with bacteria. The obtained results seem to be interesting and encourage deepening the study for other types of plants in other conditions.Keywords: constructed wetlands, macrophytes, sewage treatment, wastewater
Procedia PDF Downloads 3912555 Characterization and Evaluation of LD Slag and Fly Ash Mixture for Their Possible Utilization in Different Sectors
Authors: Jagdeep Nayak, Biswajit Paul, Anup Gupta
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Characterization of coal refuses to fly ash, and steel slag from steel industries have been performed to develop a mixture of both these materials to enhance strength properties of their utilization in other sectors like mine fill, construction work, etc. A large amount of Linz-Donawitz (LD) slag and fly ash waste are generated from steel and thermal power industries respectively. Management of these wastes is problematic, and their reutilization may provide a sustainable waste management option. LD slag and fly ash mixed in different proportions were tested to analyse the micro structural improvement and hardening rate of the matrix. Mixing of activators such as sodium hydroxide and potassium silicate with silica-alumina of LD slag-fly ash mixture, geopolymeric structure were found to be developed. The effect of geo-polymerization behaviour and subsequent structural rearrangement has been studied using compressibility; shear strength and permeability tests followed by micro-graphical analysis. Densification in the mixture was observed along with an improvement of geotechnical properties due to the addition of LD slag. Due to suitable strength characteristics of these two waste materials as mixture, it can be used in the various construction field or may be used as a filling material in mine voids.Keywords: LD slag, fly-ash, geopolymer, strength property, compressibility
Procedia PDF Downloads 3802554 Understanding the Benefits of Multiple-Use Water Systems (MUS) for Smallholder Farmers in the Rural Hills of Nepal
Authors: RAJ KUMAR G.C.
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There are tremendous opportunities to maximize smallholder farmers’ income from small-scale water resource development through micro irrigation and multiple-use water systems (MUS). MUS are an improved water management approach, developed and tested successfully by iDE that pipes water to a community both for domestic use and for agriculture using efficient micro irrigation. Different MUS models address different landscape constraints, water demand, and users’ preferences. MUS are complemented by micro irrigation kits, which were developed by iDE to enable farmers to grow high-value crops year-round and to use limited water resources efficiently. Over the last 15 years, iDE’s promotion of the MUS approach has encouraged government and other key stakeholders to invest in MUS for better planning of scarce water resources. Currently, about 60% of the cost of MUS construction is covered by the government and community. Based on iDE’s experience, a gravity-fed MUS costs approximately $125 USD per household to construct, and it can increase household income by $300 USD per year. A key element of the MUS approach is keeping farmers well linked to input supply systems and local produce collection centers, which helps to ensure that the farmers can produce a sufficient quantity of high-quality produce that earns a fair price. This process in turn creates an enabling environment for smallholders to invest in MUS and micro irrigation. Therefore, MUS should be seen as an integrated package of interventions –the end users, water sources, technologies, and the marketplace– that together enhance technical, financial, and institutional sustainability. Communities are trained to participate in sustainable water resource management as a part of the MUS planning and construction process. The MUS approach is cost-effective, improves community governance of scarce water resources, helps smallholder farmers to improve rural health and livelihoods, and promotes gender equity. MUS systems are simple to maintain and communities are trained to ensure that they can undertake minor maintenance procedures themselves. All in all, the iDE Nepal MUS offers multiple benefits and represents a practical and sustainable model of the MUS approach. Moreover, there is a growing national consensus that rural water supply systems should be designed for multiple uses, acknowledging that substantial work remains in developing national-level and local capacity and policies for scale-up.Keywords: multiple-use water systems , small scale water resources, rural livelihoods, practical and sustainable model
Procedia PDF Downloads 2802553 Effects of Alkalinity on the Treatment of Landfill Leachate through Algae Growth
Authors: Tahir Imran Qureshi
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This study was aimed at finding out effects of potential influence of alkalinity on the treatment of landfill leachate through the growth of algae at varying dilution rates and toxicity potential. pH control proved to be an effective factor influencing on algal growth. With the use of algae Scenedesmus sp. for the treatment of leachate, a sharp increase in the growth of algae was recorded until pH 9. However, at pH 9.3 and 25 °C temperature, the growing trend of algae population showed a weakening tendency with the increase of total alkalinity in the leachate solution. Highest growth of algae was recorded in the leachate samples with alkalinity ranged at 1500-2500 mg CaCO3/L under neutral condition at pH 7 after 48 hours of cultivation time. Under the similar conditions, total nitrogen and total phosphorous in the leachate also reduced to 80% and 85%, respectively, however, no significant removal of COD was observed during the course of experiment.Keywords: leachate treatment, microalgae, nutrient removal, ammonia toxicity
Procedia PDF Downloads 3252552 Cloning and Expression of the ansZ Gene from Bacillus sp. CH11 Isolated from Chilca salterns in Peru
Authors: Stephy Saavedra, Annsy C. Arredondo, Gisele Monteiro, Adalberto Pessoa Jr, Carol N. Flores-Fernandez, Amparo I. Zavaleta
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L-asparaginase from bacterial sources is used in leukemic treatment and food industry. This enzyme is classified based on its affinity towards L-asparagine and L-glutamine. Likewise, ansZ genes express L-asparaginase with higher affinity to L-asparagine. The aim of this work was to clone and express of ansZ gene from Bacillus sp. CH11 isolated from Chilca salterns in Peru. The gene encoding L-asparaginase was cloned into pET15b vector and transformed in Escherichia coli BL21 (DE3) pLysS. The expression was carried out in a batch culture using LB broth and 0.5 mM IPTG. The recombinant L-asparaginase showed a molecular weight of ~ 39 kDa by SDS PAGE and a specific activity of 3.19 IU/mg of protein. The cloning and expression of ansZ gene from this halotolerant Bacillus sp. CH11 allowed having a biological input to improve a future scaling-up.Keywords: ansZ gene, Bacillus sp, Chilca salterns, recombinant L-asparaginase
Procedia PDF Downloads 1692551 Antioxidant Capacity and Total Phenolic Content of Aqueous Acetone and Ethanol Extract of Edible Parts of Moringa oleifera and Sesbania grandiflora
Authors: Perumal Siddhuraju, Arumugam Abirami, Gunasekaran Nagarani, Marimuthu Sangeethapriya
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Aqueous ethanol and aqueous acetone extracts of Moringa oleifera (outer pericarp of immature fruit and flower) and Sesbania grandiflora white variety (flower and leaf) were examined for radical scavenging capacities and antioxidant activities. Ethanol extract of S. grandiflora (flower and leaf) and acetone extract of M. oleifera (outer pericarp of immature fruit and flower) contained relatively higher levels of total dietary phenolics than the other extracts. The antioxidant potential of the extracts were assessed by employing different in vitro assays such as reducing power assay, DPPH˙, ABTS˙+ and ˙OH radical scavenging capacities, antihemolytic assay by hydrogen peroxide induced method and metal chelating ability. Though all the extracts exhibited dose dependent reducing power activity, acetone extract of all the samples were found to have more hydrogen donating ability in DPPH˙ (2.3% - 65.03%) and hydroxyl radical scavenging systems (21.6% - 77.4%) than the ethanol extracts. The potential of multiple antioxidant activity was evident as it possessed antihemolytic activity (43.2 % to 68.0 %) and metal ion chelating potency (45.16 - 104.26 mg EDTA/g sample). The result indicate that acetone extract of M. oleifera (OPIF and flower) and S. grandiflora (flower and leaf) endowed with polyphenols, could be utilized as natural antioxidants/nutraceuticals.Keywords: antioxidant activity, Moringa oleifera, polyphenolics, Sesbania grandiflora, underutilized vegetables
Procedia PDF Downloads 3312550 ICAM1 Expression is Enhanced by TNFa through Histone Methylation in Human Brain Microvessel Cells
Authors: Ji-Young Choi, Jungjin Kim, Sang-Sun Yun, Sangmee Ahn Jo
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Intracellular adhesion molecule1 (ICAM1) is a mediator of inflammation and involved in adhesion and transmigration of leukocytes to endothelial cells, resulting in enhancement of brain inflammation. We hypothesized that increase of ICAM1 expression in endothelial cells is an early step in the pathogenesis of brain diseases such as Alzheimer’s disease. Here, we report that ICAM1 expression is regulated by pro-inflammatory cytokine TNFa in human microvascular endothelial cell (HBMVEC). TNFa significantly increased ICAM1 mRNA and protein levels at the concentrations showing no cell toxicity. This increase was also shown in micro vessels of mouse brain 24 hours after treatment with TNFa (8 mg/kg, i.v). We then investigated the epigenetic mechanism involved in the induction of ICAM1 expression. Chromatin immunoprecipitation assay revealed that TNFa reduced methylation of histone3K9 (H3K9-2me) and histone3K27 (H3K27-3me), well-known modification as gene suppression, with in the ICAM1 promoter region. However, acetylation of H3K9 and H3K14, well-known modification as gene activation, was not changed by TNFa. Treatment of BIX01294, a specific inhibitor of histone methyltransferase G9a responsible for H3K9-2me, dramatically increased in ICAM1 mRNA and protein levels and overexpression of G9a gene suppressed TNFa-induced ICAM1 expression. In contrast, GSK126, an inhibitor of histone methyltransferase EZH2 responsible for H3K27-3me and valproic acid, an inhibitor of histone deacetylase (HDAC) did not affect ICAM1 expression. These results suggested that histone3 methylation is involved in ICAM1 repression. Moreover, TNFa or BIX01294-induced ICAM induction resulted in both enhancements in adhesion and transmigration of leukocyte on endothelial cell. This study demonstrates that TNFa upregulates ICAM1 expression through H3K9-2me and H3K27-3me within the ICAM1 promoter region, in which G9a is likely to play a pivotal role in ICAM1 transcription. Our study provides a novel mechanism for ICAM1 transcription regulation in HBMVEC.Keywords: ICAM1, TNFa, HBMVEC, H3K9-2me
Procedia PDF Downloads 3242549 Effect of Yogurt Bacteria and Probiotics on Melatonin Content and Antioxidant Activity of Chickpea Yogurt
Authors: T. Utaida, A. Moongngarm
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Chickpea yogurt, a distinct dairy-free alternative, has gained popularity as a nutritious and promising future food product. Chickpea is considered as alternative protein sources because of its high protein content, suitable chemical composition for the growth of lactic acid bacteria, and antioxidant properties. Therefore, this study determined the impact of the co‐culture of yogurt bacteria with probiotics on the melatonin content, antioxidant properties, and quality characteristics of chickpea yogurt compared with traditional dairy yogurt. Chickpea yogurt was prepared using four different combinations of yogurt bacteria and probiotics including (1) Lactobacillus bulgaricus and Streptococcus thermophilus (CP-YC; control), (2) Bifidobacterium lactis, L. acidophilus, L.bulgaricus and S.thermophilus (CP-BY) (3) B.lactis, L.acidophilus and S. thermophilus (CP-BT) and (4) L. acidophilus, L. bulgaricus and S. thermophiles (CP-LA). The highest content of melatonin and tryptophan (2.05 and 711.85 ng/g dry weight, respectively) were observed in the CP-BT and CP-LA. Moreover, chickpea yogurt fermented by CP-BY exhibited a strong antioxidant activity evaluated by DPPH and ABTS assay while the CP-LA was high in FRAP assay. The pH of all treatments was decreased while the content of lactic acid was increased with fermentation time (8 h). The syneresis (%), color values (L*, a*, b*), and texture profile analysis of all treatments were significantly affected by bacteria yogurt. This study suggests that chickpea yogurt fermented by bacteria yogurt mixed with probiotics has potential for enhancing melatonin content and antioxidant activity. However, future research is required to investigate yogurt's sensory attributes, microbial stability, and shelf life.Keywords: antioxidants, dairy-free alternative, health benefits, plant-based protein, tryptophan.
Procedia PDF Downloads 12548 Study on Influencing Factors of Walkability of Rail Transit Station Area
Authors: Yang Wenjuan, Xu Yilun
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Based on the comparative analysis of the relevant evaluation methods of walking environment, this paper selects the combined evaluation method of macro urban morphology analysis and micro urban design quality survey, then investigates and analyzes the walking environment of three rail transit station area in Nanjing to explore the influence factor and internal relation of walkability of rail transit station area. Analysis shows that micro urban design factors have greater impact on the walkability of rail transit station area compared with macro urban morphology factors, the convenience is the key factor in the four aspects of convenience, security, identity and comfortability of the urban design factors, the convenience is not only affected by the block network form, but also related to the quality of the street space. The overall evaluation of walkability comes from the overlapping and regrouping of the walking environment at different levels, but some environmental factors play a leading role. The social attributes of pedestrians also partly influence their walking perception and evaluation.Keywords: rail transit station area, walkability, evaluation, influence factors
Procedia PDF Downloads 2862547 Processing and Characterization of Glass-Epoxy Composites Filled with Linz-Donawitz (LD) Slag
Authors: Pravat Ranjan Pati, Alok Satapathy
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Linz-Donawitz (LD) slag a major solid waste generated in huge quantities during steel making. It comes from slag formers such as burned lime/dolomite and from oxidizing of silica, iron etc. while refining the iron into steel in the LD furnace. Although a number of ways for its utilization have been suggested, its potential as a filler material in polymeric matrices has not yet been explored. The present work reports the possible use of this waste in glass fiber reinforced epoxy composites as a filler material. Hybrid composites consisting of bi-directional e-glass-fiber reinforced epoxy filled with different LD slag content (0, 7.5, 15, 22.5 wt%) are prepared by simple hand lay-up technique. The composites are characterized in regard to their density, porosity, micro-hardness and strength properties. X-ray diffractography is carried out in order to ascertain the various phases present in LDS. This work shows that LD slag, in spite of being a waste, possesses fairly good filler characteristics as it modifies the strength properties and improves the composite micro-hardness of the polymeric resin.Keywords: characterization, glass-epoxy composites, LD slag, waste utilization
Procedia PDF Downloads 3812546 Cement Matrix Obtained with Recycled Aggregates and Micro/Nanosilica Admixtures
Authors: C. Mazilu, D. P. Georgescu, A. Apostu, R. Deju
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Cement mortars and concretes are some of the most used construction materials in the world, global cement production being expected to grow to approx. 5 billion tons, until 2030. But, cement is an energy intensive material, the cement industry being responsible for cca. 7% of the world's CO2 emissions. Also, natural aggregates represent non-renewable resources, exhaustible, which must be used efficiently. A way to reduce the negative impact on the environment is the use of additional hydraulically active materials, as a partial substitute for cement in mortars and concretes and/or the use of recycled concrete aggregates (RCA) for the recovery of construction waste, according to EU Directive 2018/851. One of the most effective active hydraulic admixtures is microsilica and more recently, with the technological development on a nanometric scale, nanosilica. Studies carried out in recent years have shown that the introduction of SiO2 nanoparticles into cement matrix improves the properties, even compared to microsilica. This is due to the very small size of the nanosilica particles (<100nm) and the very large specific surface, which helps to accelerate cement hydration and acts as a nucleating agent to generate even more calcium hydrosilicate which densifies and compacts the structure. The cementitious compositions containing recycled concrete aggregates (RCA) present, in generally, inferior properties compared to those obtained with natural aggregates. Depending on the degree of replacement of natural aggregate, decreases the workability of mortars and concretes with RAC, decrease mechanical resistances and increase drying shrinkage; all being determined, in particular, by the presence to the old mortar attached to the original aggregate from the RAC, which makes its porosity high and the mixture of components to require more water for preparation. The present study aims to use micro and nanosilica for increase the performance of some mortars and concretes obtained with RCA. The research focused on two types of cementitious systems: a special mortar composition used for encapsulating Low Level radioactive Waste (LLW); a composition of structural concrete, class C30/37, with the combination of exposure classes XC4+XF1 and settlement class S4. The mortar was made with 100% recycled aggregate, 0-5 mm sort and in the case of concrete, 30% recycled aggregate was used for 4-8 and 8-16 sorts, according to EN 206, Annex E. The recycled aggregate was obtained from a specially made concrete for this study, which after 28 days was crushed with the help of a Retsch jaw crusher and further separated by sieving on granulometric sorters. The partial replacement of cement was done progressively, in the case of the mortar composition, with microsilica (3, 6, 9, 12, 15% wt.), nanosilica (0.75, 1.5, 2.25% wt.), respectively mixtures of micro and nanosilica. The optimal combination of silica, from the point of view of mechanical resistance, was later used also in the case of the concrete composition. For the chosen cementitious compositions, the influence of micro and/or nanosilica on the properties in the fresh state (workability, rheological characteristics) and hardened state (mechanical resistance, water absorption, freeze-thaw resistance, etc.) is highlighted.Keywords: cement, recycled concrete aggregates, micro/nanosilica, durability
Procedia PDF Downloads 572545 Anti -proliferative and Apoptotic Effects of Selected Saudi Herbs from the Rhamnaceae, Polygonaceae, and Apocynaceae Families Against Various Cancer Cell Lines
Authors: Allulu Yousef Alturki, Raghad Abdullah Alshafi, Sara Abdulaziz Alghashem, Sahar Saleh Alghamdi, Rasha Saad Suliman, Zeyad Alehaideb, Rizwan Ali
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Cancer is recognized as a worldwide public health concern. Therefore, there is a continuous quest to discover new effective medications with less side-effects. In recent years, researchers have shown an increased interest in medicinal plants as several plant species have shown promising biological activities. Thus, we seek to investigate three medicinal herbs that are commonly-found in the Middle Easternregion and yet have not been explored in depth, including plants belonging to the Rhamnaceae, Polygonaceae, and Apocynaceaeplant families. Initially, we investigated using three types of cancer cell lines for breast, colorectal, and liver cancers. We performed high Content Imaging (HCI)-Apoptosis Assay and ApoTox-Glo™ Triplex Assay on KAIMRC2 and HCT8 cell lines. The highest activity of HCI-Apoptosis Assay was with Calligonumcomosum and Ziziphusnummularia in ethanol, followed by Calotropis procera and Ziziphusnummularia in ethyl acetate. The IC50values for the families of Rhamnaceae, Polygonaceae, and Apocynaceae in HepG2 and HCT8 cell lines ranged from 0.089 to 9.84mg/mL and 0.080to 15.08mg/mL, respectively. Further screening was conducted on an additional two cell lines, namely the MDA-MB-231 and KAIMRC2, for selected seven extracts with the highest activity having IC50values ranged from 0.058 to0.51mg/mL and 0.029 to0.19mg/mL, respectively. Continuous scientific investigations to isolate and characterize the potent bioactive phytochemical(s) are warranted. Funding: The authors acknowledge financial support from King Abdullah International Medical Research Center (KAIMRC), Ministry of National Guard Health Affairs, Riyadh, Kingdom of Saudi Arabia. Institutional Review Board Statement: The study was approved by the Institutional Review Board of the Institutional Review Board of King Abdullah International Medical Research Center (SP21R/463/12, 24 January 2022). Acknowledgments: The authors want to express their gratitude to the College of Pharmacy (COP) at King Saud bin Abdulaziz University for Health Sciences (KSAU-HS) and King Abdullah International Medical Research Center (KAIMRC) for their continued support.Keywords: rhamnaceae, polygonaceae, apocynaceae, natural products
Procedia PDF Downloads 1062544 Nutritional Composition of Selected Wild Fruits from Minna Area of Niger State, Nigeria
Authors: John O. Jacob, Abdullahi Mann, Olanrewaju I. Adeshina, Mohammed M. Ndamitso
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Strychnos spinosa, Detarium microcarpum, Diospyros mespiliformis, Dialium guineese and Gardenia ternifolia are some of the wild fruits consume in the villages around Minna, Niger State. This investigation was conducted to assess the nutritional potentials of these fruits both for human consumption and for possible application in animal feed formulations. Standard analytical methods were employed in the determination of the various nutritional parameters. The proximate analysis results showed that the moisture contents ranged between (6.17-10.70%); crude fat (2.04-8.85%); crude protein (5.16-6.80%); crude fibre (7.23-19.65%); Ash (3.46-5.56%); carbohydrate (57.77-69.79%); energy value (284.49-407 kcal/mg); Vitamin C (7.2-39.93 mg/100g). The mineral analysis shows that the selected wild fruits could contribute considerable amount of both micro and macro elements to human nutrition potassium, sodium and calcium range between; potassium (343.27-764.71%); sodium (155.04-348.44%); calcium (52.47-101%). The macro element for the fruits pulp were in the order K>Na>Mg>Ca, hence, they could be included in diet to supplement daily nutrient requirement and in animal feed formulations. The domestication of these fruits is also encouraged.Keywords: mineral, micro-elements, macro-elements, feed suppleme
Procedia PDF Downloads 4332543 Assessment of Biofilm Production Capacity of Industrially Important Bacteria under Electroinductive Conditions
Authors: Omolola Ojetayo, Emmanuel Garuba, Obinna Ajunwa, Abiodun A. Onilude
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Introduction: Biofilm is a functional community of microorganisms that are associated with a surface or an interface. These adherent cells become embedded within an extracellular matrix composed of polymeric substances, i.e., biofilms refer to biological deposits consisting of both microbes and their extracellular products on biotic and abiotic surfaces. Despite their detrimental effects in medicine, biofilms as natural cell immobilization have found several applications in biotechnology, such as in the treatment of wastewater, bioremediation and biodegradation, desulfurization of gas, and conversion of agro-derived materials into alcohols and organic acids. The means of enhancing immobilized cells have been chemical-inductive, and this affects the medium composition and final product. Physical factors including electrical, magnetic, and electromagnetic flux have shown potential for enhancing biofilms depending on the bacterial species, nature, and intensity of emitted signals, the duration of exposure, and substratum used. However, the concept of cell immobilisation by electrical and magnetic induction is still underexplored. Methods: To assess the effects of physical factors on biofilm formation, six American typed culture collection (Acetobacter aceti ATCC15973, Pseudomonas aeruginosa ATCC9027, Serratia marcescens ATCC14756, Gluconobacter oxydans ATCC19357, Rhodobacter sphaeroides ATCC17023, and Bacillus subtilis ATCC6633) were used. Standard culture techniques for bacterial cells were adopted. Natural autoimmobilisation potentials of test bacteria were carried out by simple biofilms ring formation on tubes, while crystal violet binding assay techniques were adopted in the characterisation of biofilm quantity. Electroinduction of bacterial cells by direct current (DC) application in cell broth, static magnetic field exposure, and electromagnetic flux were carried out, and autoimmobilisation of cells in a biofilm pattern was determined on various substrata tested, including wood, glass, steel, polyvinylchloride (PVC) and polyethylene terephthalate. Biot Savart law was used in quantifying magnetic field intensity, and statistical analyses of data obtained were carried out using the analyses of variance (ANOVA) as well as other statistical tools. Results: Biofilm formation by the selected test bacteria was enhanced by the physical factors applied. Electromagnetic induction had the greatest effect on biofilm formation, with magnetic induction producing the least effect across all substrata used. Microbial cell-cell communication could be a possible means via which physical signals affected the cells in a polarisable manner. Conclusion: The enhancement of biofilm formation by bacteria using physical factors has shown that their inherent capability as a cell immobilization method can be further optimised for industrial applications. A possible relationship between the presence of voltage-dependent channels, mechanosensitive channels, and bacterial biofilms could shed more light on this phenomenon.Keywords: bacteria, biofilm, cell immobilization, electromagnetic induction, substrata
Procedia PDF Downloads 1822542 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 2772541 Development of Excellent Water-Repellent Coatings for Metallic and Ceramic Surfaces
Authors: Aditya Kumar
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One of the most fascinating properties of various insects and plant surfaces in nature is their water-repellent (superhydrophobicity) capability. The nature offers new insights to learn and replicate the same in designing artificial superhydrophobic structures for a wide range of applications such as micro-fluidics, micro-electronics, textiles, self-cleaning surfaces, anti-corrosion, anti-fingerprint, oil/water separation, etc. In general, artificial superhydrophobic surfaces are synthesized by creating roughness and then treating the surface with low surface energy materials. In this work, various super-hydrophobic coatings on metallic surfaces (aluminum, steel, copper, steel mesh) were synthesized by chemical etching process using different etchants and fatty acid. Also, SiO2 nano/micro-particles embedded polyethylene, polystyrene, and poly(methyl methacrylate) superhydrophobic coatings were synthesized on glass substrates. Also, the effect of process parameters such as etching time, etchant concentration, and particle concentration on wettability was studied. To know the applications of the coatings, surface morphology, contact angle, self-cleaning, corrosion-resistance, and water-repellent characteristics were investigated at various conditions. Furthermore, durabilities of coatings were also studied by performing thermal, ultra-violet, and mechanical stability tests. The surface morphology confirms the creation of rough microstructures by chemical etching or by embedding particles, and the contact angle measurements reveal the superhydrophobic nature. Experimentally it is found that the coatings have excellent self-cleaning, anti-corrosion and water-repellent nature. These coatings also withstand mechanical disturbances such surface bending, adhesive peeling, and abrasion. Coatings are also found to be thermal and ultra-violet stable. Additionally, coatings are also reproducible. Hence aforesaid durable superhydrophobic surfaces have many potential industrial applications.Keywords: superhydrophobic, water-repellent, anti-corrosion, self-cleaning
Procedia PDF Downloads 2892540 Access of Small and Medium Enterprises to Finance in Rural Areas: Case of Indonesia and Thailand
Authors: N. Ikasari, T. Sumransat, U. Eko, R. Kusumastuti
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Small and medium enterprises (SMEs) are regarded as the engine for economic development, notwithstanding their continuous financing conundrum. In the case of developing countries, access to finance is a reflection of the effectiveness of government policy. The widely accepted perspective to assess small businesses’ access to finance is that of economic view. The existing body of literature presents access to finance in three dimensions; they are accessibility, eligibility and affordability. Within this perspective, the role of socio-cultural has not explored. This study is aimed at investigating the existence of any socio-cultural factors within access to finance issue in Asian countries where governance is enriched by countries’ values and beliefs. The significance of this study is the instigation of supplementary dimension to assess access to finance that eventually contributes to the development of micro-finance policy. Indonesia and Thailand are selected as cases in point, where distinction is drawn on the level of cultural diversity and micro-finance policy in respective country. A questionnaire is used to collect information related to the three dimensions of access to finance as well as to explore alternative financing reasoning to elaborate the issue from the demand side. Questionnaires are distributed to 60 small business owners operating in Indonesia and the same number in Thailand. In order to present a complete understanding on the matter at hand, interviews with banks are conducted to capture the perspective as presented by the supply side. Research findings show that small business owners and banks in Indonesia and Thailand are in agreement that access to finance is not deemed as an issue. However, trust issue that exists mutually between financing users and providers leads small business owners in Indonesia to look for alternative financing other than banks. The findings contribute to the refinement of micro-financing policy in Indonesia and Thailand.Keywords: access to finance, Indonesia, small and medium enterprises, Thailand
Procedia PDF Downloads 2842539 Epigenetic Mechanisms Involved in the Occurrence and Development of Infectious Diseases
Authors: Frank Boris Feutmba Keutchou, Saurelle Fabienne Bieghan Same, Verelle Elsa Fogang Pokam, Charles Ursula Metapi Meikeu, Angel Marilyne Messop Nzomo, Ousman Tamgue
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Infectious diseases are one of the most important causes of morbidity and mortality worldwide. These diseases are caused by micro-pathogenic organisms, such as bacteria, viruses, parasites, and fungi. Heritable changes in gene expression that do not involve changes to the underlying DNA sequence are referred to as epigenetics. Emerging evidence suggests that epigenetic mechanisms are important in the emergence and progression of infectious diseases. Pathogens can manipulate host epigenetic machinery to promote their own replication and evade immune responses. The Human Genome Project has provided new opportunities for developing better tools for the diagnosis and identification of target genes. Several epigenetic modifications, such as DNA methylation, histone modifications, and non-coding RNA expression, have been shown to influence infectious disease outcomes. Understanding the epigenetic mechanisms underlying infectious diseases may result in the progression of new therapeutic approaches focusing on host-pathogen interactions. The goal of this study is to show how different infectious agents interact with host cells after infection.Keywords: epigenetic, infectious disease, micro-pathogenic organism, phenotype
Procedia PDF Downloads 712538 A Novel Upregulated circ_0032746 on Sponging with MIR4270 Promotes the Proliferation and Migration of Esophageal Squamous Cell Carcinoma
Authors: Sachin Mulmi Shrestha, Xin Fang, Hui Ye, Lihua Ren, Qinghua Ji, Ruihua Shi
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Background: Esophageal squamous cell carcinoma (ESCC) is a tumor arising from esophageal epithelial cells and is one of the major disease subtype in Asian countries, including China. Esophageal cancer is the 7th highest incidence based on the 2020 data of GLOBOCAN. The pathogenesis of cancer is still not well understood as many molecular and genetic basis of esophageal carcinogenesis has yet to be clearly elucidated. Circular RNAs are RNA molecules that are formed by back-splicing covalently joined 3′- and 5′-endsrather than canonical splicing, and recent data suggest circular RNAs could sponge miRNAs and are enriched with functional miRNA binding sites. Hence, we studied the mechanism of circular RNA, its biological function, and the relationship between microRNA in the carcinogenesis of ESCC. Methods: 4 pairs of normal and esophageal cancer tissues were collected in Zhongda hospital, affiliated to Southeast University, and high-throughput RNA sequencing was done. The result revealed that circ_0032746 was upregulated, and thus we selected circ_0032746 for further study. The backsplice junction of circRNA was validated by sanger sequence, and stability was determined by RNASE R assay. The binding site of circRNA and microRNA was predicted by circinteractome,mirandaand RNAhybrid database. Furthermore, circRNA was silenced by siRNA and then by lentivirus. The regulatory axis of circ0032746/miR4270 was validated by shRNA, mimic, and inhibitor transfection. Then, in vitro experiments were performed to assess the role of circ0032746 on proliferation (CCK-8 assay and colon formation assay), migration and invasion (Transewell assay), and apoptosis of ESCC. Results: The upregulated circ0032746 was validated in 9 pairs of tissues and 5 types of cell lines by qPCR, which showed high expression and was statistically significant (P<0.005) ). Upregulated circ0032746 was silenced by shRNA, which showed significant knockdown in KYSE 30 and TE-1 cell lines expression compared to control. Nuclear and cytoplasmic mRNA fraction experiment displayed the cytoplasmic location of circ0032746. The sponging of miR4270 was validated by co-transfection of sh-circ0032746 and mimic or inhibitor. Transfection with mimic showed the decreased expression of circ_0032746, whereas inhibitor inhibited the result. In vitro experiments showed that silencing of circ_0032746 inhibited the proliferation, migration, and invasion compared to the negative control group. The apoptosis was seen higher in a knockdown group than in the control group. Furthermore, 11 common mircoRNA target mRNAs were predicted by Targetscan, MirTarbase, and miRanda database, which may further play role in the pathogenesis. Conclusion: Our results showed that novel circ_0032746 is upregulated in ESCC and plays role in itsoncogenicity. Silencing of circ_0032746 inhibits the proliferation and migration of ESCC whereas increases the apoptosis of cancer cells. Hence, circ0032746 acts as an oncogene in ESCC by sponging with miR4270 and could be a potential biomarker in the diagnosis of ESCC in the future.Keywords: circRNA, esophageal squamous cell carcinoma, microRNA, upregulated
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