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The study of metals mobility derived from mining activities in an ultramafic lithology is limited. This study investigates the effects of distance on potentially toxic metals such as Co, Cu, Fe, Mn, Ni, Pb, and Zn pollution, and the geochemical processes of fluvial system downstream of an ex-copper mine (Mamut River). The toxicity level of the river was evaluated using various sediment quality guidelines, ecotoxicological risks (ecological risk and risk index) and pollution indices. The geochemical behavior and stability of these toxic metals in the solid-phase samples were also examined. The results show that elevated concentrations of Ni, Cu, and Fe in the sediments can be linked to the adsorption and precipitation of metals from the aqueous-phase samples. We found that the metal scavenging rate as a function of distance is more evident in tropical environments than it was previously thought (10 km downstream). Such an inference could be explained by the greater amount of rainfall (pH 5.5-6.5) received in the tropics and higher weathering products that could react and form stable complexes. Geochemical analysis of the river sediment indicates that Ni, Cu, and Fe in the river sediment have increased 44-, 81-, and 90-fold compared to the background values, respectively. A significant decrease in the concentration of the potentially toxic metals was found at 5.5 km downstream. The scavenging rate of Fe is the highest (1485.82 µg g-1 km-1) followed by Cu (141.48 µg g-1 km-1), Ni (10.23 µg g-1 km-1), Pb (8.12 µg g-1 km-1) and Zn (5.01 µg g-1 km-1) in the tropical river system. In contrast, the concentration of Co and Mn in the river sediments doubled as the river flows approximately 5 km downstream due to the higher mineral solubility and weaker metal partition coefficient. This study also discusses the possibility of asbestos (mainly as chrysotile in the X-ray diffraction) as a potential hidden risk present within the ultramafic setting. This case study can be extrapolated to explain the dispersion of inorganic pollutants in an ultramafic environment in a global context.
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Metais Pesados , Poluentes Químicos da Água , China , Monitoramento Ambiental , Sedimentos Geológicos , Chumbo , Medição de Risco , RiosRESUMO
The research study was carried out to evaluate trace metals (Pb, Cd, Se, Al, Mn, Cu, Zn, Fe, As, Ni, Cr, and Ag) concentrations in groundwater of Lorong Serai 4, Hulu Langat, Selangor, Malaysia. Additionally, the research study focused on determining non-carcinogenic and carcinogenic health risks, sources of the contaminants, and effective remediation methods. The results show that the concentration levels of Pb, Cd, Se, Al, Cu, Zn, Ni, Cr, and Ag are lower than their corresponding permissible limits, while Fe, Mn, and As concentrations exceed their acceptable limit. The hazard index of the groundwater in the area exceeded the acceptable limit, showing the rate of carcinogenic and non-carcinogenic health effects associated with the water. The findings also indicate that the lifetime cancer risk is high compared to the maximum limits of lifetime cancer risk from the drinking water (10-6 to 10-4). The groundwater geochemical data of the area are used in establishing the source of Fe, Mn, and As metal ions. Evaluation of Fe2+/Fe3+ and S2-/SO42- redox couples and thermodynamic modelling indicates that the groundwater of the area is in redox disequilibrium. The groundwater samples contain aqueous iron sulphate, which is supersaturated, ferrous carbonate and aluminium sulphate that are saturated. The main state of redox disequilibrium is governed by mineral precipitation and dissolution. Aqueous arsenic and manganese are possibly derived from the dissolution of pyrite (arsenopyrite) and amorphous oxide-hydroxides, respectively. The high concentration of iron in the shallow groundwater in the area is primarily the result of silicate rock weathering of ferroan igneous and metamorphic minerals with a minor contribution from the oxidation of iron sulphides. Magnetite coated with graphene oxide (Fe3O4-GO) nanoparticles (NPs) was synthesized and characterized, and the adsorption preliminary experiments were carried out; and the Fe3O4-GO NPs show enhanced removal (Fe > As > Mn) capacity over graphene oxide (GO).
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Recuperação e Remediação Ambiental , Metais/análise , Oligoelementos/análise , Poluentes Químicos da Água/análise , Adsorção , Monitoramento Ambiental , MalásiaRESUMO
The toxicity and kinetic uptake potential of zinc oxide (ZnO) and titanium dioxide (TiO2) nanomaterials into the red bean (Vigna angularis) plant were investigated. The results obtained revealed that ZnO, due to its high dissolution and strong binding capacity, readily accumulated in the root tissues and significantly inhibited the physiological activity of the plant. However, TiO2 had a positive effect on plant physiology, resulting in promoted growth. The results of biochemical experiments implied that ZnO, through the generation of oxidative stress, significantly reduced the chlorophyll content, carotenoids and activity of stress-controlling enzymes. On the contrary, no negative biochemical impact was observed in plants treated with TiO2. For the kinetic uptake and transport study, we designed two exposure systems in which ZnO and TiO2 were exposed to red bean seedlings individually or in a mixture approach. The results showed that in single metal oxide treatments, the uptake and transport increased with increasing exposure period from one week to three weeks. However, in the metal oxide co-exposure treatment, due to complexation and competition among the particles, the uptake and transport were remarkably decreased. This suggested that the kinetic transport pattern of the metal oxide mixtures varied compared to those of its individual constituents.
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Nanopartículas Metálicas/toxicidade , Titânio/toxicidade , Testes de Toxicidade , Vigna/efeitos dos fármacos , Óxido de Zinco/toxicidade , Cinética , Nanopartículas Metálicas/química , Estresse Oxidativo , Vigna/fisiologia , Óxido de Zinco/químicaRESUMO
This study explores the possible application of a biodegradable plant based surfactant, obtained from Sapindus mukorossi, for washing low levels of arsenic (As) from an iron (Fe) rich soil. Natural association of As(V) with Fe(III) makes the process difficult. Soapnut solution was compared to anionic surfactant sodium dodecyl sulfate (SDS) in down-flow and a newly introduced suction mode for soil column washing. It was observed that soapnut attained up to 86% efficiency with respect to SDS in removing As. Full factorial design of experiment revealed a very good fit of data. The suction mode generated up to 83 kPa pressure inside column whilst down-flow mode generated a much higher pressure of 214 kPa, thus making the suction mode more efficient. Micellar solubilisation was found to be responsible for As desorption from the soil and it followed 1st order kinetics. Desorption rate coefficient of suction mode was found to be in the range of 0.005 to 0.01, much higher than down-flow mode values. Analysis of the FT-IR data suggested that the soapnut solution did not interact chemically with As, offering an option for reusing the surfactant. Soapnut can be considered as a soil washing agent for removing As even from soil with high Fe content.
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Arsênio/química , Oxirredutases/química , Sapindus/química , Poluentes do Solo/química , Solo/química , Tensoativos/química , CinéticaRESUMO
Bestari Jaya, former tin mining catchment covers an area of 2656.31 hectares comprised of four hundred and forty-two different-size lakes and ponds. The present study area comprise of 92 hectares of the catchment that include four large size lakes. Arc GIS version 9.2 used to develop bathymetric map, Global Positioning System (GPS) for hydrographical survey and flow meter was utilized for water discharge analysis (flow routing) of the catchment. The water quality parameters (pH, temperature, electric conductivity, dissolved oxygen DO, total dissolved solids TDS, chlorides, ammonium, nitrates) were analyzed by using Hydrolab. Quality assurance (QA) and quality control (QC) procedures were strictly followed throughout the field work and data analysis. Different procedures were employed to evaluate the analytical data and to check for possible transcription or dilution errors, changes during analysis, or unusual or unlikely values. The results obtained are compared with interim national water quality standards for Malaysia indicates that water quality of area is highly degraded. It is concluded that Bestri Jaya ex-mining catchment has a high pollution potential due to mining activities and River Ayer Hitam, recipient of catchment water, is a highly polluted river.
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Área Programática de Saúde , Resíduos Industriais/análise , Mineração , Oxigênio/análise , Estanho/análise , Poluentes Químicos da Água/análise , Água/química , Condutividade Elétrica , Concentração de Íons de Hidrogênio , Malásia , Água/análiseRESUMO
Early depletion of carbon-based electron donors and pollution swapping are among the recalcitrant challenges for in situ remediation technology. There is a pressing need to provide a slow but steady dose of materials for a prolonged period in order to sustain in situ treatments. In an effort to develop effective sequence multi-barrier reactive media that can address these challenges, the present study reports laboratory column experiments using organic (date seed, Moringa oleifera and wood chips) and inorganic (limestone; CaCO3) carbonaceous materials in varying proportions. The substances are supported by paleo sandy beach soil and are sequentially packed in line with the geometric pattern of a hypothetical funnel and gate for nitrate (NO3 -) remediation. The optimized reactive media show remarkable efficiency, remediating 94.4% of NO3 - as a target pollutant within the system. They also remediate 99.20% of the NO3 - down the gradient and 96.4% of its lethal by-product - NO2 -. However, with the generation of an 11-fold biomass build-up, the system is only able to eliminate 53% of the total organic carbon generated. Hence, the advective dispersive features of aquifers are required for the complete remediation of the generated secondary biomass. The results also reveal that the efficiency of the sequential media depends on the composition/ratio of the constituents, flow rate and the initial concentration of NO3 - in the influent. Surface characterization studies (FESEM-EDX and FTIR) show that compared to wood chips, date seed is less prone to degradation, suggesting a longer life span. A degradation study affirms the findings of the surface studies as it reveals that the half-life of date seed is three times longer than that of wood chips. In addition, the study reveals that the double exponential model is more suitable than the single exponential model in determining the rate of decay of both wood chips and date seed.
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Carbon based materials are emerging as a sustainable alternative to their metal-oxide counterparts. However, their transport behavior under natural aqueous environment is poorly understood. This study investigated the transport and retention profiles of carbon nanoparticles (CNPs) and graphene oxide quantum dots (GOQDs) through column experiments in saturated porous media. CNPs and GOQDs (30â¯mg/L) were dispersed in natural river water (RW) and passed through the column at a flow rate of 1â¯mL/min, which mimicking the natural water flow rate. After every 10â¯min, the column effluents were collected and the mass recovery and retention profiles were monitored. Results indicated that the transport of both carbonaceous colloids was predominantly controlled by surface potential and ionic composition of natural water. The CNPs with its high surface potential (-40â¯mV) exhibited more column transport and was less susceptible to solution pH (5.6-6.8) variation as compared to GOQDs (-24â¯mV). The results showed that, monovalent salt (NaCl) was one of the dominating factors for the retention and transport of carbonaceous colloids compared to divalent salt (CaCl2). Furthermore, the presence of natural organic matter (NOM) increased the transport of both carbonaceous colloids and thereby decreases the tendency for column retention.
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Carbono/química , Coloides/química , Água/química , Cromatografia/métodos , Grafite/química , Nanopartículas , Compostos Orgânicos/análise , Concentração Osmolar , Pontos Quânticos , Cloreto de Sódio/análiseRESUMO
A comprehensive study was conducted to identify the salinization origins and the major hydrogeochemical processes controlling the salinization and deterioration of the Gaza coastal aquifer system through a combination approaches of statistical and geostatistical techniques, and detailed hydrogeochemical assessments. These analyses were applied on ten physicochemical variables for 219 wells using STATA/SE12 and Surfer softwares. Geostatistical analysis of the groundwater salinity showed that seawater intrusion along the coastline, and saltwater up-coning inland highly influenced the groundwater salinity of the study area. The hierarchical cluster analysis (HCA) technique yielded seven distinct hydrogeochemical signature clusters; (C1&C2: Eocene brackish water invasion, C3 saltwater up-coning, C4 human inputs, C5 seawater intrusion, C6 & C7 rainfall and mixing inputs). Box plot shows a wide variation of most of the ions while Chadha's plot elucidates the predominance of Na-Cl (71.6%) and Ca/Mg-Cl (25%) water types. It is found that, the highest and the lowest levels of salinization and the highest level of nitrate pollution were recorded in the northern area. This result reflects the sensitivity of this area to the human activities and/or natural actions. Around 90.4% of the wells are nitrate polluted. The main source of nitrate pollution is the sewage inputs while the farming inputs are very limited and restricted mostly in the sensitive northern area. Among the hydrogeochemical processes, ion exchange process was the most effective process all over the study area. Carbonate dissolution was common in the study area with the highest level in clusters 6, 7, 4 and 2 in the north while Gypsum dissolution was significant only in cluster 1 in the south and limited in the other clusters. This integrated multi-techniques research should be of benefit for effective utilization and management of the Gaza coastal aquifer system as well as for future work in other similar aquifers systems.
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Monitoramento Ambiental , Água Subterrânea/química , Salinidade , Abastecimento de Água/estatística & dados numéricos , Oriente Médio , Poluentes Químicos da Água/análise , Poços de ÁguaRESUMO
Presently, engineered nanomaterials (ENMs) are used in a wide variety of commercial applications, resulting in an uncontrolled introduction into the aquatic environment. The purpose of this review is to summarize the pathways and factors that controlling the transport and toxicity of five extensively used ENMs. These toxicological pathways are of great importance and need to be addressed for sustainable implications of ENMs without environmental liabilities. Here we discuss five potentially utilized ENMs with their possible toxicological risk factors to aquatic plants, vertebrates model and microbes. Moreover, the key effect of ENMs surface transformations by significant reaction with environmental objects such as dissolved natural organic matter (DOM) and the effect of ENMs surface coating and surface charge will also be debated. The transformations of ENMs are subsequently facing a major ecological transition that is expected to create a substantial toxicological effect towards the ecosystem. These transformations largely involve chemical and physical processes, which depend on the properties of both ENMs and the receiving medium. In this review article, the critical issues that controlling the transport and toxicity of ENMs are reviewed by exploiting the latest reports and future directions and targets are keenly discussed to minimize the pessimistic effects of ENMs.
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Rare earth elements (REEs) are becoming significant due to their huge applications in many industries, large-scale mining and refining activities. Increasing usage of such metals pose negative environmental impacts. In this research ICP-MS has been used to analyze soil samples collected from former ex-mining areas in the depths of 0-20 cm, 21-40 cm, and 41-60 cm of residential, mining, natural, and industrial areas of Perak. Principal component analysis (PCA) revealed that soil samples taken from different mining, industrial, residential, and natural areas are separated into four clusters. It was observed that REEs were abundant in most of the samples from mining areas. Concentration of the rare elements decrease in general as we move from surface soil to deeper soils.
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Monitoramento Ambiental , Metais Terras Raras/análise , Poluentes do Solo/análise , Solo/química , MineraçãoRESUMO
Experiments on hybrid red tilapia Oreochromis sp. were conducted to assess histopathological effects induced in gill tissues of 96 h exposure to waterborne lead (5.5 mg/L). These tissues were investigated by light and scanning electron microscopy. Results showed that structural design of gill tissues was noticeably disrupted. Major symptoms were changes of epithelial cells, fusion in adjacent secondary lamellae, hypertrophy and hyperplasia of chloride cells and coagulate necrosis in pavement cells with disappearance of its microridges. Electron microscopic X-ray microanalysis of fish gills exposed to sublethal lead revealed that lead accumulated on the surface of the gill lamella. This study confirmed that lead exposure incited a difference of histological impairment in fish, supporting environmental watch over aquatic systems when polluted by lead.
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A study was carried out to determine the level of rare earth elements (REEs) in water and sediment samples from ex-mining lakes and River in Kinta Valley, Perak, Malaysia. Surface water and sediments from an ex-mining lake and Kinta River water samples were analyzed for REEs by inductively coupled plasma mass spectrometry. The total concentration of REEs in the ex-mining lake water samples and sediments were found to be 3685 mg/l and 14159 mg/kg, respectively, while the total concentration of REEs in Kinta River water sample was found to be 1224 mg/l. REEs in mining lake water were found to be within 2.42 mg/l (Tb) to 46.50 mg/l (Ce), while for the Kinta River, it was 1.33 mg/l (Ho) to 29.95 mg/l (Ce). Sediment samples were also found with REEs from 9.81 mg/kg (Ho) to 765.84 mg/kg (Ce). Ce showed the highest average concentrations for mining lake (3.88 to 49.08 mg/l) and Kinta River (4.44 to 33.15 mg/l) water samples, while the concentration of La was the highest (11.59 to 771.61 mg/kg) in the mining lake sediment. Lu was shown to have the highest enrichment of REEs in ex-mining lake sediments (107.3). Multivariate statistical analyses such as factor analysis and principal component analysis indicated that REEs were associated and controlled by mixed origin, with similar contributions from anthropogenic and geogenic sources. The speciation study of REEs in ex-tin mining sediments using a modified five-stage sequential extraction procedure indicated that yttrium (Y), gadolinium (Gd), and lanthanum (La) were obtained at higher percentages from the adsorbed/exchanged/carbonate fraction. The average potential mobility of the REEs was arranged in a descending order: Yb > Gd > Y = Dy > Pr > Er > Tm > Eu > Nd > Tb > Sc > Lu > Ce > La, implying that under favorable conditions, these REEs could be released and subsequently pollute the environment.
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Metais Terras Raras/análise , Mineração , Poluentes do Solo/análise , Estanho/análise , Poluentes Químicos da Água/análise , Sedimentos Geológicos/química , Lagos/química , Malásia , Rios/químicaRESUMO
A series of doped and un-doped magnetic adsorbents CuCexFe2-xO4 (x=0.0-0.5) for fluoride were prepared with the micro-emulsion method. Fluoride adsorption was optimized for solution pH, temperature, contact time, and initial concentration and was monitored via normal phase ion chromatography (IC). The effect of concomitant anions was also explored to perform and simulate competitive fluoride adsorption in real water samples. Optimal adsorption was discovered by a simple quadratic model based on central composite design (CCD) and the response surface method (RSM). The adsorption, electrochemical and magnetic properties were compared between doped and un-doped ferrites. Doped ferrites (x=0.1-0.5) were found to be superior to un-doped ferrites (x=0) regarding the active sites, functional groups and fluoride adsorption. The characterization, optimization and application results of the doped ferrites indicated enhanced fluoride adsorption and easy separation with a simple magnet.
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Heavy-metal-contaminated soil is one of the major environmental pollution issues all over the world. In this study, two low-cost amendments, inorganic eggshell and organic banana stem, were applied to slightly alkaline soil for the purpose of in situ immobilization of Pb, Cd, and Zn. The artificially metal-contaminated soil was treated with 5% eggshell or 10% banana stem. To simulate the rainfall conditions, a metal leaching experiment for a period of 12 weeks was designed, and the total concentrations of the metals in the leachates were determined every 2 weeks. The results from the metal leaching analysis revealed that eggshell amendment generally reduced the concentrations of Pb, Cd, and Zn in the leachates, whereas banana stem amendment was effective only on the reduction of Cd concentration in the leachates. A sequential extraction analysis was carried out at the end of the experiment to find out the speciation of the heavy metals in the amended soils. Eggshell amendment notably decreased mobility of Pb, Cd, and Zn in the soil by transforming their readily available forms to less accessible fractions. Banana stem amendment also reduced exchangeable form of Cd and increased its residual form in the soil.
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Casca de Ovo/química , Recuperação e Remediação Ambiental/métodos , Metais Pesados/análise , Musa/química , Poluentes do Solo/análise , Animais , Cádmio/análise , Fracionamento Químico , Chumbo/análise , Caules de Planta/química , Solo/química , Solo/normas , Água/química , Zinco/análiseRESUMO
Methyl esters from vegetable oils have attracted a great deal of interest as substitute for petrodiesel to reduce dependence on imported petroleum and provide an alternate and sustainable source for fuel with more benign environmental properties. In the present study biodiesel was prepared from sunflower seed oil by transesterification by alkali-catalyzed methanolysis. The fuel properties of sunflower oil biodiesel were determined and discussed in the light of ASTM D6751 standards for biodiesel. The sunflower oil biodiesel was chemically characterized with analytical techniques like FT-IR, and NMR ((1)H and (13)C). The chemical composition of sunflower oil biodiesel was determined by GC-MS. Various fatty acid methyl esters (FAMEs) were identified by retention time data and verified by mass fragmentation patterns. The percentage conversion of triglycerides to the corresponding methyl esters determined by (1)H NMR was 87.33% which was quite in good agreement with the practically observed yield of 85.1%.
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Field and laboratory studies were conducted to estimate concentration of potential contaminants from landfill in the underlying groundwater, leachate, and surface water. Samples collected in the vicinity of the landfill were analyzed for physiochemical parameters, organic contaminants, and toxic heavy metals. Water quality results obtained were compared from published data and reports. The results indicate serious groundwater and surface water contamination in and around the waste disposal site. Analysis of the organic samples revealed that the site contains polychlorinated biphenyls and other organo-chlorine chemicals, principally chloro-benzenes. Although the amount of PCB concentration discovered was not extreme, their presence indicates a potentially serious environmental threat. Elevated concentrations of lead, copper, nickel, manganese, cadmium, and cobalt at the downgradient indicate that the contamination plume migrated further from the site, and the distribution of metals and metals containing wastes in the site is nonhomogeneous. These results clearly indicate that materials are poorly contained and are at risk of entering the environment. Therefore, full characterization of the dump contents and the integrity of the site are necessary to evaluate the scope of the problem and to identify suitable remediation options.
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Monitoramento Ambiental/métodos , Eliminação de Resíduos/métodos , Poluentes Químicos da Água/análise , Fenômenos Químicos , Água Subterrânea/análise , Metais Pesados/análise , Bifenilos Policlorados/análise , Rios/química , Poluição da Água/análiseRESUMO
The potential of three submerged aquatic plant species (Cabomba piauhyensis, Egeria densa, and Hydrilla verticillata) to be used for As, Al, and Zn phytoremediation was tested. The plants were exposed for 14 days under hydroponic conditions to mine waste water effluents in order to assess the suitability of the aquatic plants to remediate elevated multi-metals concentrations in mine waste water. The results show that the E. densa and H. verticillata are able to accumulate high amount of arsenic (95.2%) and zinc (93.7%) and resulted in a decrease of arsenic and zinc in the ambient water. On the other hand, C. piauhyensis shows remarkable aluminium accumulation in plant biomass (83.8%) compared to the other tested plants. The ability of these plants to accumulate the studied metals and survive throughout the experiment demonstrates the potential of these plants to remediate metal enriched water especially for mine drainage effluent. Among the three tested aquatic plants, H. verticillata was found to be the most applicable (84.5%) and suitable plant species to phytoremediate elevated metals and metalloid in mine related waste water.
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Organismos Aquáticos/metabolismo , Biodegradação Ambiental , Hydrocharitaceae/metabolismo , Alumínio/metabolismo , Organismos Aquáticos/química , Arsênio/química , Arsênio/metabolismo , Ouro/química , Ouro/metabolismo , Hydrocharitaceae/química , Águas Residuárias/química , Água/química , Zinco/química , Zinco/metabolismoRESUMO
BACKGROUND: Renilla Luciferase reporter gene (rLuc) GL4.82 and GL4.13 promoter are key player in transfection, but precise knowledge of its targets in colon cancer remains limited. The aim of this study was to characterize the best transfection technique to produce a stable transfected colon DLD1 (colorectal adenocarcinoma cell line), therefore imaging based approaches were employed. RESULTS: DLD1 cells were transfected with a Plasmid (SV40-RLuc) carrying Renilla luciferase under the control of the SV-40 promoter, by using two different transfection techniques. Cells expressing the required DNA were isolated after antibiotic (Puramycin) selection. Clones of DLD-1/SV40-RLuc were produced using two different techniques (96 well plates and Petri dish) and their florescence intensity was recorded using IVIS machine (Calliper Life Sciences, Hopkinton, USA). Both techniques were characterized with the help of serial dilution technique. Results from this study substantiated that electroporation is the best. As expected, clones varied in their specific luciferase activity along with the dilutions. With the increase in cell concentration increase in intensity of florescence was recorded. CONCLUSIONS: Based on the results we are confident that this transfected cell line DLD-1/SV40-RLuc (colorectal adenocarcinoma cell line) is the best for further Orthotopic Xenotransplantation Studies and in-vivo experiments as well. Investigation shows that DLD1/SV-rLuc cells have gained little bit resistance against both drugs therefore further study is suggested to know the reasons.
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This research was conducted to evaluate the combination of electromigration and potassium permanganate as a potential remediation method for low-permeability media (e.g., soil and sediment) contaminated with dissolved and sorbed organic contaminants. The experimental procedure was composed of two stages: determination of migration rates of permanganate through homogeneous cores and a primarily qualitative analysis of migration in more heterogeneous, two-dimensional scenarios. Results indicated that transport of permanganate through fine-grained porous media and clays can be undertaken using electromigration, and electromigration rates were found to be at least 400% faster than diffusion alone. In addition, the use of an applied electric field in a flushing scenario was shown to result in almost 100% sweep efficiency of a domain consisting of clay blocks interspersed in a glass bead medium. The results of the study show that there is potential for this method to be able to deliver permanganate and other potential remedial agents to treat contaminated zones within heterogeneous and low-permeability porous media through in situ chemical oxidation or other processes.