Search results for: Acid mine drainage

Authors: Supannika Klangphukhiew, Roongnapa Srichana, Rina Patramanon

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Cortisol has been used as a well-known commercial stress biomarker. A homeostasis response to psychological stress is indicated by an increased level of cortisol produced in hypothalamus-pituitary-adrenal (HPA) axis. Chronic psychological stress contributing to the high level of cortisol relates to several health problems. In this study, the cortisol biosensor was fabricated that mimicked the natural receptors. The artificial antibodies were prepared using molecular imprinted polymer technique that can imitate the performance of natural anti-cortisol antibody with high stability. Cortisol-molecular imprinted polymer (cortisol-MIP) was obtained using the multi-step swelling and polymerization protocol with cortisol as a target molecule combining methacrylic acid:acrylamide (2:1) with bisacryloyl-1,2-dihydroxy-1,2-ethylenediamine and ethylenedioxy-N-methylamphetamine as cross-linkers. Cortisol-MIP was integrated to the sensor. It was coated on the disposable screen-printed carbon electrode (SPCE) for portable electrochemical analysis. The physical properties of Cortisol-MIP were characterized by means of electron microscope techniques. The binding characteristics were evaluated via covalent patterns changing in FTIR spectra which were related to voltammetry response. The performance of cortisol-MIP modified SPCE was investigated in terms of detection range, high selectivity with a detection limit of 1.28 ng/ml. The disposable cortisol biosensor represented an application of MIP technique to recognize steroids according to their structures with feasibility and cost-effectiveness that can be developed to use in point-of-care.

Keywords: stress biomarker, cortisol, molecular imprinted polymer, screen-printed carbon electrode

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Authors: Prima Luna, Afroditi Chatzifragkou, Dimitris Charalampopoulos

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Sorghum byproducts, namely bran, stalk, and panicle are examples of lignocellulosic biomass. These raw materials contain large amounts of polysaccharides, in particular hemicelluloses, celluloses, and lignins, which if efficiently extracted, can be utilised for the development of a range of added value products with potential applications in agriculture and food packaging sectors. The aim of this study was to characterise fractions extracted from sorghum bran and stalk with regards to their physicochemical properties that could determine their applicability as food-packaging materials. A sequential alkaline extraction was applied for the isolation of cellulosic, hemicellulosic and lignin fractions from sorghum stalk and bran. Lignin content, phenolic content and antioxidant capacity were also investigated in the case of the lignin fraction. Thermal analysis using differential scanning calorimetry (DSC) and X-Ray Diffraction (XRD) revealed that the glass transition temperature (T_g) of cellulose fraction of the stalk was ~78.33 ^oC at amorphous state (~65%) and water content of ~5%. In terms of hemicellulose, the T_g value of stalk was slightly lower compared to bran at amorphous state (~54%) and had less water content (~2%). It is evident that hemicelluloses generally showed a lower thermal stability compared to cellulose, probably due to their lack of crystallinity. Additionally, bran had higher arabinose-to-xylose ratio (0.82) than the stalk, a fact that indicated its low crystallinity. Furthermore, lignin fraction had T_g value of ~93 ^oC at amorphous state (~11%). Stalk-derived lignin fraction contained more phenolic compounds (mainly consisting of p-coumaric and ferulic acid) and had higher lignin content and antioxidant capacity compared to bran-derived lignin fraction.

Keywords: alkaline extraction, bran, cellulose, hemicellulose, lignin, stalk

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Authors: I. Hilia, C. Hange, F. Hakala, M. Matheus, C. Jansen, J. Hidinwa, O. Awofolu

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The article reports on the occurrences, level, and mobility of selected trace metals in environmental samples. The conceptual basis was to examine the possible influence of anthropogenic activities and the impact on human and environmental health. Environmental samples (soil, plant and lower animal) were randomly collected from stratified study/sampling areas, preserved and pre-treated before analysis. Mineral acid digestion procedure was employed for the isolation of metallic contents in samples, and elemental qualitative and quantitative analysis was by ICP-OES. Analytical protocol was validated through the quality assurance process and was found acceptable with quantitative metallic recoveries in the range of 85-90%; hence considered applicable for the analyses of environmental samples. The mean concentration of analysed metals in soil samples ranged from 53.2- 2532.8 mg/kg (Cu); 59.5- 2020.1 mg/kg (Zn); 1.80 – 21.26 mg/kg (Cd) and 19.6- 140.9 mg/kg (Pb). The mean level in grass samples ranged from 9.33 – 38.63 mg/kg (Cu); 64.20-105.18 mg/kg (Zn); 0.28–0.73 mg/kg (Cd) and 0.53 -16.26 mg/kg (Pb) while the mean level in lower animal sample (beetle) varied from 9.6 - 105.3 mg/kg (Cu); 134.1-297.2 mg/kg (Zn); 0.63 – 3.78 (Cd) and 8.0 – 29.1 mg/kg (Pb) across sample collection points (SCPs) 1-4 respectively. Metallic transfer factors (TFs) were in the order Zn >Cd > Cu > Pb with metal Pollution Indices (MPIs) in the order SCP1 > SCP2 > SCP3 > SCP4. About 60-70 % of analysed metals were above the maximum allowable limits (MALs) in soil and plant samples. Results obtained revealed the general prevalence of analysed metals at all sampled sites with indication of metallic mobility across the food chain which portrayed dire consequences for environmental and human health. Systematic environmental remediation and pollution abatement strategies are recommended.

Keywords: trace metals, pollution, human health, Incidences, ICP-OES

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Authors: K. B. M. Islam, Syeeda Shiraj-Um-Mahmuda, Afroj Jahan, A. A. Bhuiyan

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In Bangladesh, the use of imported probiotics in poultry is gradually being increased. But surprisingly, no probiotic bacteria have been isolated yet in Bangladesh despite the existence of scavenging native poultry as potential source that is seemingly more resistant to GIT infection as well as other diseases. Therefore, the study was undertaken to isolate, identify and characterize the potential probiotic Lactobacillus and Bifidobacteria strains from Bangladeshi indigenous poultry, and to evaluate their suitability to use in poultry industry. Crop and cecal samples from 61 healthy indigenous birds were used to isolate potential probiotics strains following conventional cultural methods. A total of 216 isolates were identified following physical, biochemical and molecular methods that belonged to the genus Lactobacillus and Bifidobacteria. An auto-aggregation test was performed for 180 and 136 isolated lactobacilli and bifidobacteria strains, respectively. Twelve lactobacilli isolates and 7 bifidobacteria isolates were selected because of their convenient aggregation. In vitro tests including antibacterial activity, resistance to low pH, hemolytic activities etc. were performed for evaluation of probiotic potential of each strain. Under the in vitro conditions and with respects to the probiotic traits, three lactobacilli; LS16, LS45, LS133 and two bifidobacteria, BS21 and BS90 were found to be potential probiotic strains. Thus, they are proposed to be evaluated for their in vivo probiotic properties. If the proposed strains are found suitable as the probiotics to be used in commercial poultry industry, it is expected that the local probiotics would be more beneficial and would save the huge amount of money that Bangladesh spends every year for the importation of such materials from abroad.

Keywords: Bangladeshi poultry, gut microbiota, lactic acid bacteria, scavenging chicken, GIT health

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Authors: Dileep Kumar Verma, Sunil Kumar Lal

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Influenza still remains one of the most challenging diseases posing significant threat to public health causing seasonal epidemics and pandemics. Influenza A Virus (IAV) surface protein hemagglutinin is known to play an important role in viral attachment to the host sialic acid receptors and concentrate in lipid rafts for efficient viral fusion. Selective nature of Influenza A virus to utilize rafts micro-domain for efficient virus assembly and budding has been explored in depth. However, the detailed mechanism of IAV binding to host cell membrane and entry into the host remains elusive. In the present study we investigated the role of lipid rafts in early life cycle events of IAV. Role of host lipid rafts was studied using raft disruption method by extraction of cholesterol by Methyl-β-Cyclodextrin. Using GM1, a well-known lipid raft marker, we were able to observe co-localization of IAV on lipid rafts on the host cell membrane. This experiment suggests a direct involvement of lipid rafts in the initiation of the IAV life cycle. Upon disruption of lipid rafts by Methyl-b-cyclodextrin, we observed a significant reduction in IAV binding on the host cell surface indicating a significant decrease in virus attachment to coherent membrane rafts. Our results provide proof that host lipid rafts and their constituents play an important role in the adsorption of IAV. This study opens a new avenues in IAV virus-host interactions to combat infection at a very early steps of the viral lifecycle.

Keywords: lipid raft, adsorption, cholesterol, methyl-β-cyclodextrin, GM1

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Authors: Seyed Mostafa Jafari Raad, Hassan Hassanzadeh

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Injection of impurities along with CO₂ into saline aquifers provides an exceptional prospect for low-cost carbon capture and storage technologies and can potentially accelerate large-scale implementation of geological storage of CO₂. We have conducted linear stability analyses and numerical simulations to investigate the effects of permitted impurities in CO₂ streams on the onset of natural convection and dynamics of subsequent convective mixing. We have shown that the rate of dissolution of an impure CO₂ stream with H₂S highly depends on the operating conditions such as temperature, pressure, and composition of impurity. Contrary to findings of previous studies, our results show that an impurity such as H₂S can potentially reduce the onset time of natural convection and can accelerate the subsequent convective mixing. However, at the later times, the rate of convective dissolution is adversely affected by the impurities. Therefore, the injection of an impure CO₂ stream can be engineered to improve the rate of dissolution of CO₂, which leads to higher storage security and efficiency. Accordingly, we have identified the most favorable CO₂ stream compositions based on the geophysical properties of target aquifers. Information related to the onset of natural convection such as the scaling relations and the most favorable operating conditions for CO₂ storage developed in this study are important in proper design, site screening, characterization and safety of geological storage. This information can be used to either identify future geological candidates for acid gas disposal or reviewing the current operating conditions of licensed injection sites.

Keywords: CO₂ storage, solubility trapping, convective dissolution, storage efficiency

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Authors: Ja’Afar Umar, Naziru Salisu

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The consumption of powdered or industrialized juices has increased globally due to the fast pace of city life. These foods, with their attractive color, odor, and taste, are easily diluted in water and can lead to obesity, diabetes, hypertension, and cardiovascular problems. In a study, 80 purple varieties of onion bulbs were used to evaluate the cytotoxicity of the Tiara and Bevi mix beverage powder. The viability of the bulbs was tested using the A. cepa toxicity test. The bulbs were divided into five groups, and the root growth was recorded. The mixture was then squashed in a 45% acetic acid solution and examined for chromosomal abnormalities. The chromosomal abnormalities were classified as bridges, c-mitoses, vagrants, fragments, stickiness, bi-nuclei, and multi-polar. The study found that the highest number of dividing cells was in the negative control group, followed by the group treated with BM beverage. The highest number of aberrant cells was in the group treated with TR beverage, followed by BM 5%. Stickiness of cells was observed in both BM and TR 5% beverage concentrations. No lagging chromosome was present in the negative control group. The highest mitotic index was in the negative control group, and bridge fragrance was observed in the groups treated with different beverages. This study highlights the importance of Allium cepa L. in genotoxic substance testing, revealing chromosomal and mitotic abnormalities in root tip cells. The study also reveals that at 5% concentrations, root growth decreases, indicating potential genetic abnormalities in Allium cepa's genetic material.

Keywords: cytotoxicity, Allium cepa, Beverages, Chromosome

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Authors: Kulvinder Bajwa, Narsi R. Bishnoi

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Finding a sustainable alternative for today’s petrochemical industry is a major challenge facing by researchers, scientists, chemical engineers, and society at the global level. Microorganisms are considered to be sustainable feedstock for 3rd generation biofuel production. In this study, we have investigated the potential of a native bacterial strain isolated from a petrol contaminated site for the production of biodiesel. The bacterium was identified to be Rhodococcus opacus by biochemical test and 16S rRNA. Compositional analysis of bacterial biomass has been carried out by Fourier transform infrared spectroscopy (FTIR) in order to confirm lipid profile. Lipid and biomass were optimized by combination with Box Behnken design (BBD) of response surface methodology. The factors selected for the optimization of growth condition were glucose, yeast extract, and ammonium nitrate concentration. The experimental model developed through RSM in terms of effective operational factors (BBD) was found to be suitable to describe the lipid and biomass production, which indicated higher lipid and biomass with a minimum concentration of ammonium nitrate, yeast extract, and quite higher dose of glucose supplementation. Optimum results of the experiments were found to be 2.88 gL⁻¹ biomass and lipid content 38.75% at glucose 20 gL⁻¹, ammonium nitrate 0.5 gL⁻¹ and yeast extract 1.25 gL⁻¹. Furthermore, GCMS study revealed that Rhodococcus opacus has favorable fatty acid profile for biodiesel production.

Keywords: biofuel, Oleaginious bacteria, Rhodococcus opacus, FTIR, BBD, free fatty acids

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Authors: E. K. Tripsanas, D. Spanos, I. Oikonomopoulos, K. Stathopoulou, A. S. Abdelsamad, A. Pagoulatos

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Patraikos Gulf is located offshore western Greece, and it is limited to the west by the Zante, Cephalonia, and Lefkas islands. The Plio/Pleistocene sequence is characterized by two depocenters, the east and west Patraikos basins separated from each other by a prominent sill. This study is based on the Plio/Pleistocene seismic stratigraphy analysis of a newly acquired 3D PSDM (Pre-Stack depth migration) seismic survey in the west Patraikos Basin and few 2D seismic profiles throughout the entire Patraikos Gulf. The eastern Patraikos Basin, although completely buried today with water depths less than 100 m, it was a deep basin during Pliocene ( > 2 km of Pliocene-Pleistocene sediments) and appears to have gathered most of Achelous River discharges. The west Patraikos Gulf was shallower ( < 1300 m of Pliocene-Pleistocene sediments) and characterized by a hummocky relief due to thrust-belt tectonics and Miocene to Pleistocene halokinetic processes. The transition from Pliocene to Miocene is expressed by a widespread erosional unconformity with evidence of fluvial drainage patterns. This indicates that west Patraikos Basin was aerially exposed during the Messinian Salinity Crisis. Continuous to semi-continuous, parallel reflections in the lower, early- to mid-Pliocene seismic packet provides evidence that the re-connection of the Mediterranean Sea with the Atlantic Ocean during Zanclean resulted in the flooding of the west Patraikos basin and the domination of hemipelagic sedimentation interrupted by occasional gravity flows. This is evident in amplitude and semblance horizon slices, which clearly show the presence of long-running, meandering submarine channels sourced from the southeast (northwest Peloponnese) and north. The long-running nature of the submarine channels suggests mobile efficient turbidity currents, probably due to the participation of a sufficient amount of clay minerals in their suspended load. The upper seismic section in the study area mainly consists of several successions of clinoforms, interpreted as progradational delta complexes of Achelous River. This sudden change from marine to shallow marine sedimentary processes is attributed to climatic changes and eustatic perturbations since late Pliocene onwards (~ 2.6 Ma) and/or a switch of Achelous River from the east Patraikos Basin to the west Patraikos Basin. The deltaic seismic unit consists of four delta complexes. The first two complexes result in the infill of topographic depressions and smoothing of an initial hummocky bathymetry. The distribution of the upper two delta complexes is controlled by compensational stacking. Amplitude and semblance horizon slices depict the development of several almost straight and short (a few km long) distributary submarine channels at the delta slopes and proximal prodeltaic plains with lobate sand-sheet deposits at their mouths. Such channels are interpreted to result from low-efficiency turbidity currents with low content in clay minerals. Such a differentiation in the nature of the gravity flows is attributed to the switch of the sediment supply from clay-rich sediments derived from the draining of flysch formations of the Ionian and Gavrovo zones, to the draining of poor in clay minerals carbonate formations of Gavrovo zone through the Achelous River.

Keywords: sequence stratigraphy, basin analysis, river deltas, submarine channels

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Authors: Layal Karam, Rayan Roustom, Mohamad G. Abiad, Tahra El-Obeid, Ioannis N. Savvaidis

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The demand for marinated chicken worldwide, is continuously growing. To date, limited data on addition of active components of Essential Oils (EOs) to marinades for chicken preservation are available. The antimicrobial effect of carvacrol and thymol, added at 0.4 and 0.8% v/w to marinated fresh chicken, stored in air and under vacuum packaging (VP), for 21 days at 4°C, was examined. The samples were monitored for microbiological (total viable count (TVC), lactic acid bacteria (LAB), Brochothrix thermosphacta, Pseudomonas spp., total coliforms, Escherichia coli, yeasts and molds) and sensory attributes (odor characteristics). Our data supports that among the tested microorganisms, Pseudomonas spp., LAB and B. thermosphacta were the most dominant microbiota in the marinated chicken samples. Additionally, the use of active EOs components, especially the higher concentration (0.8% v/w) in combination with VP, retarded the growth of spoilage microbiota and resulted in a significant reduction of about 2.9-3.1 log cfu/g and a microbiological shelf-life extension of marinated chicken by > 6 days, as judged by TVC data. Interestingly, the combination of active components of EOs at the lower concentration (0.4% v/w) and packaging (air or vacuum) resulted in a significant sensorial shelf-life extension of 15 and >21 days, as compared to the controls’ shelf-life of 9 days. The results of our study demonstrated the potential of the active components, carvacrol and thymol, as natural effective antimicrobial hurdles to control the growth of spoilage microorganisms in marinated chicken meat.

Keywords: chicken, essential oils compounds, marination, meat spoilage, preservation

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Authors: Alireza Barari, Ahmad Abdi

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The aim of this study was to determine the effects of the effects of six weeks endurance training and Aloe Vera on cyclooxygenase 2 (COX-2) and VEGF levels in mice with breast cancer. For this purpose, 35 rats were randomly divided into 5 groups: control (healthy), control (cancer), training (cancer), Aloe Vera (cancer) and Aloe Vera + training (cancer). Induction of breast cancer tumors were done in mice by planting method. The training program includes six weeks of swimming training was done in three sessions per week. Training time from 10 minutes on the first day increased to 60 minutes in second week, and by stabilizing this time, the water flow rate was increased from 7 to 15 liters per minute. 300 mg per kg body weight of Aloe Vera extract was injected into the peritoneal. Sampling was done 48 hours after the last exercise session. K-S test to determine the normality of the data and analysis of variance for repeated measures and Tukey test was used to analyze the data. A significant difference in the p<0.05 accepted. The results showed that induction of cancer cells significantly increased levels of COX-2 in aloe group and VEGF in training and Aloe Vera + training groups. The results suggest that swimming exercise and Aloe Vera can reduce levels of COX-2 and VEGF in mice with breast cancer.The results of this study, Induction of cancer cells significantly increased levels of COX-2 and MMP-9 in the control group compared with the cancer control group. The results suggest that Aloe Vera can probably inhibit the cyclooxygenase pathway and thus production of prostaglandin E2 decrease of arachidonic acid.

Keywords: endurance training, aloe vera, COX-2, VEGF

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Authors: Muhammad Yasir, Debarun Dutta, Mark Willcox

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Biomaterial-associated infections are a multi-billion dollar burden globally. Antimicrobial peptide-based coatings may be able to prevent such infections. The aim of this study was to investigate the mechanism of action surface bound peptides (AMPs) against Pseudomonas aeruginosa 6294. Melimine and Mel4 were covalently attached to glass coverslips using azido-benzoic acid. Attachment was confirmed using X-ray photoelectron spectroscopy. P. aeruginosa was allowed to attach to AMP-coated glass for up to 6 hours. The effect of the surface-bound AMPs on bacterial cell membranes was evaluated using the dyes DiSC3-(5), Sytox green, SYTO 9 and propidium iodide with fluorescence microscopy. Release of cytoplasmic materials ATP and DNA/RNA were determined in the surrounding fluid. The amount of cell death was estimated by agar plate counts. The AMPs were successfully covalently bound to the glass as demonstrated by increases in %nitrogen of 3.6% (melimine) and 2.3% (Mel4) compared to controls. Immobilized peptides disrupted the cytoplasmic membrane potential of P. aeruginosa within 10 min. This was followed by the release of ATP after 2 h. Membrane permeabilization started at 3 h of contact with glass coated AMPs. There was a significant number of bacteria (59% for melimine; 36% for Mel-4) with damaged membranes after 4 h of contact. At the 6 h time point, release of DNA occurred with melimine releasing 2 times the amount of DNA/RNA than Mel4 surfaces (p < 0.05). Surface bound AMPs were able to disrupt cell membranes with subsequent release of cytoplasmic materials, and ultimately resulting in bacterial death.

Keywords: biomaterials, immobilized antimicrobial peptides, P. aeruginosa, mode of action

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Authors: A. O. Adebayo, G. A. Idowu, F. Odegbemi

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Investigations on the recovery of nickel from aqueous solution using 2-hydroxy-5-nonyl- acetophenone oxime (LIX-84I) impregnated on activated charcoal was carried out. The LIX-84I was impregnated onto the pores of dried activated charcoal by dry method and optimum conditions for different equilibrium parameters (pH, adsorbent dosage, extractant concentration, agitation time and temperature) were determined using a simulated solution of nickel. The kinetics and adsorption isotherm studies were also evaluated. It was observed that the efficiency of recovery with LIX-84I impregnated on charcoal was dependent on the pH of the aqueous solution as there was little or no recovery at pH below 4. However, as the pH was raised, percentage recovery increases and peaked at pH 5.0. The recovery was found to increase with temperature up to 60ºC. Also it was observed that nickel adsorbed onto the loaded charcoal best at a lower concentration (0.1M) of the extractant when compared with higher concentrations. Similarly, a moderately low dosage (1 g) of the adsorbent showed better recovery than larger dosages. These optimum conditions were used to recover nickel from the leachate of Ni-MH batteries dissolved with sulphuric acid, and a 99.6% recovery was attained. Adsorption isotherm studies showed that the equilibrium data fitted best to Temkin model, with a negative value of constant, b (-1.017 J/mol) and a high correlation coefficient, R² of 0.9913. Kinetic studies showed that the adsorption process followed a pseudo-second order model. Thermodynamic parameter values (∆G⁰, ∆H⁰, and ∆S⁰) showed that the adsorption was endothermic and spontaneous. The impregnated charcoal appreciably recovered nickel using a relatively smaller volume of extractant than what is required in solvent extraction. Desorption studies showed that the loaded charcoal is reusable for three times, and so might be economical for nickel recovery from waste battery.

Keywords: charcoal, impregnated, LIX-84I, nickel, recovery

Procedia PDF Downloads 147

Authors: Murat Tosunoglu, Cigdem Gul, Nurcihan Hacioglu, Nurdan Tepeova

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Amphibian and Reptilian species are influenced by pollution and habitat destruction. Blood parameters of Amphibia species were particularly affected by the negative environmental conditions. Studied frog samples 36 clinically normal Pelophylax ridibundus individuals were captured along the Biga Stream between April–June 2014. When comparing our findings with the Turkish legislation (Water pollution control regulation), the 1. Locality of the Biga stream in terms of total coliform classified as "high quality water" (Coliform: 866.66 MPN/100 mL), while the 2. locality was a "contaminated water" (Coliform: 53266.66 MPN/100 mL). Blood samples of the live specimens were obtained in the laboratory within one day of their capture. The blood samples were taken from the etherized frogs by means of ventriculus punctures, via heparinized hematocrit capillaries. Hematological and biochemical analyses based on high quality water and contaminated water, respectively, are as follows: Red blood cell count (444210.52-426846.15 per cubic millimeter of blood), white blood cell count (4215.78-4684.61 per cubic millimeter of blood), hematocrit value (29.25-29.43 %), hemoglobin concentration (7.76-7.22 g/dl), mean corpuscular volume (637.64-719.99 fl), mean corpuscular hemoglobin (184.78-174.75 pg), mean corpuscular hemoglobin concentration (29.44-24.82 %), glucose (103.74-124.13 mg/dl), urea (87.68-81.72 mg/L), cholesterol (148.20-197.39 mg/dl), creatinine (0.29-0.28 mg/dl), uric acid (10.26-7.55 mg/L), albumin (1.13-1.39 g/dl), calcium (11.45-9.70 mg/dl), triglyceride (135.23-155.85 mg/dl), total protein (4.26-3.73 g/dl), phosphorus (6.83-17.86 mg/dl), and magnesium (0.95-1.06 mg/dl). The some hematological parameters in P. ridibundus specimens are given for the first time in this study. No water quality dependent variation was observed in clinic hematology parameters measured.

Keywords: Pelophylax ridibundus, hematological parameters, biochemistry, freshwater quality

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Authors: Ying Li, Zaisheng Hong, Weihong Wang

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With the rapid enlargement of the size of Chinese universities, newly built campuses are springing up everywhere in recent years. It is urged to build eco-green campus because the role of higher education institutions in the transition to a more sustainable society has been highlighted for almost three decades. On condition that a new campus is usually built on an undeveloped site, where the basic infrastructure is not completed, finding proper strategies in planning and design of the campus becomes a primary concern. Low Impact Development (LID) options have been proposed as an alternative approach to make better use of rainwater in planning and design of an undeveloped site. On the basis of analyzing the natural circumstance, geographic condition, and other relative information, four main LID approaches are coordinated in this study of Hebei Union University, which are ‘Storage’, ‘Retaining’, ‘Infiltration’ and ‘Purification’. ‘Storage’ refers to a big central lake in the campus for rainwater harvesting. ‘Retaining’ means rainwater gardens scattered in the campus, also being known as bioretention areas which mimic the naturally created pools of water, to decrease surface flow runoff. ‘Infiltration’ is designed of grassed swales, which also play a part of floodway channel. ‘Purification’ is known as either natural or artificial wetland to reduce pollutants such as nitrogen and phosphorous in the waterbody. With above mentioned measures dealing with the synthetic use of rainwater in the acid & alkali area in the coastal district, an eco-green campus construction and an ecological sustainability will be realized, which will give us more enlightenment and reference.

Keywords: newly built campus, low impact development, planning design, rainwater reuse

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Authors: Asmaa M. Fahim

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In this elucidation, we synthesized different heterocyclic compounds attached to Benzene sulfonamide moiety via (E)-N-(4-(3-(4-bromophenyl)acryloyl)phenyl)-4-methyl benzene sulfonamide which is obtained from Nucleophilic substitution reaction between 4-methylbenzene sulfonyl chloride and 1-(4-aminophenyl)ethan-1-one in pyridine to get N-(4-acetyl phenyl)-4-methyl benzenesulfonamide which reacted 4-bromobenzal dehyde undergoes aldol condensation in NaOH to afford the corresponding chalchone 4. Moreover, the reactivity of chalchone 4 showed several active methylene derivatives utilized the pressurized microwave irradiation as a green energy resource. Chalcone 4 was allowed to react with ethyl cyanoacetate and acetylacetone, respectively, at 70 °C with pressure under microwave reaction condition to afford the 5-cyano-6-oxo-1,2,5,6-tetrahydropyridin-2-yl)-4-methylbenzenesulfonamide 6 and N-(4'-acetyl-4''-bromo-5'-oxo-2',3',4',5'-tetrahydro-[1,1':3',1''-terphenyl]-4-yl)-4-methylbenzenesulfonamide 8 derivatives. Moreover, the reactivity of this sulphonamide chalchone with NH2NH2 in EtOH and acetic acid, which gave 2,5-dihydro-1H-imidazol-4-yl)-4-methyl benzenesulfonamide, 1H-pyrazol-3-yl)-4-methyl and reactivity with NH2OH.HCl gave isoxazol-3-yl)-4-methylbenzenesulfonamide derivatives. The synthesized compounds were screened for their ADME properties and directed to antitumor activity on HepG2 hepatocellular carcinoma and MCF-7 breast cancer and exhibited excellent behavior against standard drugs; these results were confirmed through molecular simulations with different proteins. Additionally, the Density Functional Theory analysis of optimized structures investigated their physical descriptors, FMO, ESP and MEP, which correlated with biological evaluation.

Keywords: synthesis, green chemistry, antitumor activity, DFT study

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Authors: Sushil Kumar Singh, Ashok Kumar, Ankit Ganeshpurkar, Ravi Singh, Devendra Kumar

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In spectrum of neurodegenerative diseases, Alzheimer’s disease (AD) is characterized by the presence of amyloid β plaques and neurofibrillary tangles in the brain. It results in cognitive and memory impairment due to loss of cholinergic neurons, which is considered to be one of the contributing factors. Donepezil, an acetylcholinesterase (AChE) inhibitor which also inhibits butyrylcholinesterase (BuChE) and improves the memory and brain’s cognitive functions, is the most successful and prescribed drug to treat the symptoms of AD. The present work is based on designing of the selective BuChE inhibitors using computational techniques. In this work, machine learning models were trained using classification algorithms followed by screening of diverse chemical library of compounds. The various molecular modelling and simulation techniques were used to obtain the virtual hits. The amide derivatives of 4-(phenylsulfonamido) benzoic acid were synthesized and characterized using 1H & 13C NMR, FTIR and mass spectrometry. The enzyme inhibition assays were performed on equine plasma BuChE and electric eel’s AChE by method developed by Ellman et al. Compounds 31, 34, 37, 42, 49, 52 and 54 were found to be active against equine BuChE. N-(2-chlorophenyl)-4-(phenylsulfonamido)benzamide and N-(2-bromophenyl)-4-(phenylsulfonamido)benzamide (compounds 34 and 37) displayed IC50 of 61.32 ± 7.21 and 42.64 ± 2.17 nM against equine plasma BuChE. Ortho-substituted derivatives were more active against BuChE. Further, the ortho-halogen and ortho-alkyl substituted derivatives were found to be most active among all with minimal AChE inhibition. The compounds were selective toward BuChE.

Keywords: Alzheimer disease, butyrylcholinesterase, machine learning, sulfonamides

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Authors: Thalía Moreno-García Malo, Santiago Torres-Ríos, María G. González-Cruz, María M. Hernández-Arroyo, Sergio R. Trejo-Estrada

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Agave atrovirens is the main source of agave sap, the raw material for the production of pulque, an artisanal fermented beverage, traditional since prehispanic times in the highlands of central Mexico. Agave sap is rich in glucose, sucrose and fructooligosaccharides, and strongly differs from agave syrup from A. tequilana, which is mostly a high molecular weight fructan. Agave sap is converted into pulque by a highly diverse microbial community which includes bacteria, yeast and even filamentous fungi. The bacterial diversity has been recently studied. But the composition of consortia derived from directed enrichments differs sharply from the whole fermentative consortium. Using classical microbiology methods, and selective liquid and solid media formulations, either bacterial or fungal consortia were developed and analyzed. Bacterial consortia able to catabolize specific prebiotic saccharides were selected and preserved for future developments. Different media formulations, selective for bacterial genera such as Bifidobacterium, Lactobacillus, Pediococcus, Lactococcus and Enterococcus were also used. For yeast, specific media, osmotic pressure and unique carbon sources were used as selective agents. Results show that most groups are represented in the enrichment cultures; although very few are recoverable from the whole consortium in artisanal pulque. Diversity and abundance vary among consortia. Potential bacterial probiotics obtained from agave sap and agave juices show tolerance to hydrochloric acid, as well as strong antimicrobial activity.

Keywords: Agave, pulque, microbial consortia, prebiotic activity

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Authors: Babar Ali, Mohammad Rashid, Showkat Rasool Mir, Mohammad Ali, Saiba Shams

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Background: The plant Camellia sinensis (Theaceae) is an evergreen shrub indigenous to Assam (India) and parts of China and Japan. Traditional Chinese medicine has recommended green tea for headaches, body aches and pains, digestion, enhancement of immune defense, detoxification, as an energizer and to prolong life. The leaves have more than 700 chemical constituents, among which flavanoids, amino acids, vitamins (C, E, K), caffeine and polysaccharides. Adulteration and substitution may affect the quality of formulation containing tea leaves. Standardization of medicinal preparation is essential for further therapeutic results and for global acceptance. Hence, chromatographic fingerprint profiles were carried out for establishing the standards. Materials and methods: TLC studies for methanolic extracts of the leaves of Camellia sinensis were carried out in a new developed solvent system, Toluene: Ethyl acetate: Formic acid (7:3:1). TLC plates were dried in air, visualized in UV at wavelengths 254 nm and 366 nm and photographed. Results: Results provide valuable clue regarding their polarity and selection of solvents for separation of phytochemicals. Fingerprinting of methanolic extract of Camellia sinensis leaves revealed the presence of various phytochemicals in UV at 254 nm and 366 nm. Conclusion: Fingerprint profile is quite helpful in setting up of standards and thus to keep a check on intentional/unintentional adulteration. TLC offers major advantages over other conventional chromatographic techniques such as unsurpassed flexibility (esp. stationary and mobile phase), choice of detection wavelength, user friendly, rapid and cost effective.

Keywords: Cammelia sinensis Linn., standardization, methanolic extract, thin layer chromatography

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Authors: Mohammad Jamal

Abstract:

Non-alcoholic fatty liver disease (NAFLD) has become one of the most chronic liver diseases, prevalent among people with morbid obesity. NAFLD does not develop clinically significant liver disease, however cirrhosis and liver cancer develop in subset and currently there are no approved therapies for the treatment of NAFLD. The study is aimed to understand the various key genes involved in the mechanism of NAFLD which can be valuable for developing diagnostic and predictive biomarkers based on their histologic stage of liver. The study was conducted on 16 male Sprague Dawley rats. The animals were divided in two groups: control group (n=8) fed on ad libitum normal chow and regular water and the cafeteria group (CAF)) (n=8) fed on high fatty/ carbohydrate diet. The animals received their respective diet from 4 weeks onwards from D.O.B until 25 weeks. Liver was extracted and RT² Profiler PCR Array was used to assess the NAFLD related genes. Histological evaluation was performed using H&E stain in liver tissue sections. Our PCR array results showed that genes involved in anti-inflammatory activity (Ifng, IL10), fatty acid uptake/oxidation (Fabp5), apoptosis (Fas), lipogenesis (Gck and Srebf1), Insulin signalling (Igfbp1) and metabolic pathway (pdk4) were upregulated in the liver of cafeteria fed obese rats. Bloated hepatocytes, displaced nucleus and higher lipid content were seen in the liver of cafeteria fed obese rats. Although Liver biopsies remain the gold standard in evaluating NAFLD, however an approach towards non-invasive markers could be used in understanding the physiology, therapeutic potential, and the targets to combat NAFLD.

Keywords: biomarkers, cafeteria diet, obesity, NAFLD

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Authors: Manisha Bal, Amit Verma, B. C. Meikap

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Carbon dioxide (CO₂) is the most abundant waste produced by human activities. It is estimated to be one of the major contributors of greenhouse effect and also considered as a major air pollutant formed by burning of fossil fuels. The main sources of emissions are flue gas from thermal power plants and process industries. It is also a contributor of acid rain. Its exposure through inhalation can lead to health risks. Therefore, control of CO₂ emission in the environment is very necessary. The main focus of this study is on the removal of carbon dioxide from off gases using a self-priming venturi scrubber in submerged conditions using sodium hydroxide as the scrubbing liquid. A self-priming submerged venturi scrubber is an efficient device to remove gaseous pollutants. In submerged condition, venturi scrubber remains submerged in the liquid tank and the liquid enters at the throat section of venturi scrubber due to the pressure difference which includes the hydrostatic pressure of the liquid and static pressure of the gas. The inlet polluted air stream enters through converging section which moves at very high velocity in the throat section and atomizes the liquid droplets. This leads to absorption of CO₂ from the off gases in scrubbing liquid which resulted in removal of CO₂ gas from the off gases. Detailed investigation on the scrubbing of carbon dioxide has been done in this literature. Experiments were conducted at different throat gas velocities, liquid levels in outer cylinder and CO₂ inlet concentrations to study the carbon dioxide removal efficiency. Experimental results give more than 95% removal efficiency of CO₂ in the self priming venturi scrubber which can meet the environmental emission limit of CO₂ to save the human life.

Keywords: carbon dioxide, scrubbing, pollution control, self-priming venturi scrubber

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Authors: Li Maksym, Prabhakar M. N., Jung-Il Song

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In this study, a flame retardant and hydrophobic cotton textile were prepared by utilizing a renewable halogen-free bio-based solution based on chitosan, urea, and phytic acid, named bio-flame retardant liquid (BFL), through facile dip-coating technology. Deposition of BFL on the surface of the cotton was confirmed by Fourier-transform infrared spectroscopy and scanning electron microscope coupled with energy-dispersive X-ray spectrometer. Thermal and flame retardant properties of the cottons were studied with thermogravimetric analysis, differential scanning calorimetry, vertical flame test, cone calorimeter test. Only with 8.8% of dry weight gain treaded cotton showed self-extinguish properties during fire test. Cone calorimeter test revealed a reduction of peak heat release rate from 203.2 to 21 kW/m2 and total heat release from 20.1 to 2.8 MJ/m2. Incidentally, BFL remarkably improved the thermal stability of flame retardant cotton from expressed in an enhanced amount of char at 700 °C (6.7 vs. 33.5%). BFL initiates the formation of phosphorous and silica contain char layer whichrestrains the propagation of heat and oxygen to unburned materialstrengthen by the liberation of non-combustible gases, which reduce the concentration of flammable volatiles and oxygen hence reducing the flammability of cotton. In addition, hydrophobicity and specific ignition test for upholstery application were performed. In conjunction, the proposed flame retardant cotton is potentially translatable to be utilized as upholstery materials in public transport.

Keywords: cotton farbic, flame retardancy, surface coating, intumescent mechanism

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Authors: Khadijat Ayanpeju Abdulsalam, Bolanle Mary Olawoye, Paul Babatunde Ayoola

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The potential of Baobab leaves is understudied and not yet fully documented. The purpose of this work is to highlight the important nutritional value and practical qualities of baobab leaves. In this research, proximate analysis was studied to determine the macronutrient quantitative analysis in baobab leaves. Studies were also conducted on other characteristics, such as moisture content, which is significant to the food business since it affects food quality, preservation, and resistance to deterioration. Dietary fiber, which was also studied, has important health benefits, such as lowering blood cholesterol levels by lowering low-density lipoprotein or "bad" cholesterol. It functions as an anti-obesity and anti-diabetic agent, lowering the likelihood of haemorrhoids developing. Additionally, increasing face bulk and short-chain fatty acid synthesis improves gastrointestinal health and overall wellness. Baobab leaves had a moisture content of 6.4%, fat of 16.1%, ash of 3.2%, protein of 18.7%, carbohydrate 57.2% and crude fiber of 4.1%. The minerals determined in the sample of baobab leaves are Ca, Fe, Mg, K, Na, P, and Zn with Potassium (347.6±0.70) as the most abundant mineral while Zn (9.31±0.60) is the least abundant. The functional properties studied include pH, gelation temperature, bulk density, water absorption capacity, oil absorption capacity, foaming property, emulsifying property, and stability and swelling capacity, which are 8.72, 29, 0.39, 138, 98.20, 0.80, 72.80, and 73.50 respectively. The Fourier Transform InfraRed absorption spectra show bands like C=O, C-Cl and N-H. Baobab leaves are edible, nutritious, and non-toxic, as the mineral contents are within the required range.

Keywords: dietary fibre, proximate analysis, macronutrients, minerals, baobab leaves, frequency range

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Authors: Asma Ansari, Afsheen Aman, Shah Ali Ul Qader

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In the past few years, numerous concerns have been raised against increased bacterial resistance towards effective drugs and become a debated issue all over the world. With the emergence of drug resistant pathogens, the interaction of natural antimicrobial compounds and antibacterial nanoparticles has emerged as a potential candidate for combating infectious diseases. Microbial diversity in the biome provides an opportunity to screen new species which are capable of producing large number of antimicrobial compounds. Among these antimicrobial compounds, bacteriocins are highly specific and efficient antagonists. A combination of bacteriocin along with nanoparticles could prove to be more potent due to broadened antibacterial spectrum with possibly lower doses. In the current study, silver nanoparticles were synthesized through biological reduction using various isolated bacterial, fungal and yeast strains. Spectroscopy and scanning electron microscopy (SEM) was performed for the confirmation of nanoparticles. Bacteriocin was characterized and purified to homogeneity through gel permeation chromatography. The estimated molecular weight of bacteriocin was 10 kDa. Amino acid analysis and N-terminal sequencing revealed the novelty of the protein. Then antibacterial potential of silver nanoparticles and broad inhibitory spectrum bacteriocin was determined through agar well diffusion assay. These synthesized bacteriocin-Nanoparticles exhibit a good potential for clinical applications as compared to bacteriocin alone. This combination of bacteriocin with nanoparticles will be used as a new sort of biocide in the field of nano-proteomics. The advancement of nanoparticles-mediated drug delivery system will open a new age for rapid eradication of pathogens from biological systems.

Keywords: BAC-IB17, multidrug resistance, purification, silver nanoparticles

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Authors: Alaa E. El-sisi, Sherin Zakaria

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Rebamipide is an antiulcer drug with unique properties such as anti-inflammatory action. It induces endogenous prostaglandin e2 (PGE2). PGE2 is considered as a potent physiological suppressor of liver fibrosis. Aim of study: This study investigated the effect of rebamipide on hepatic fibrosis. Material and Method: Hepatic fibrosis was induced by intraperitoneal injections (IP) injection of CCl4 (0.45 mL/kg) in corn oil 1:5 twice a week for 4 weeks. Rats were divided into four groups as follow: Group 1 treated with CCL4 only, group 2 and 3 treated with CCL4 and rebamipide 60 mg/kg/day (group2) or 100 mg/kg/day (group3), and the fourth group was considered as control group and treated with vehicles. ALT, AST, and Bilirubin were assayed in serum. Antioxidant markers such as malondialdhyde (MDA) and superoxide dismutase (SOD) and fibrotic markers such as hyaluronic acid (HA) and procollagen-III (procol-III) were evaluated in liver tissues. IL-10 as well as PGE2 were also assayed in liver tissues. Pathologic changes in the liver were detected by hematoxylin and eosin staining. Collagen precipitation in liver tissues was visualized using masson trichrom stain. Results: Rebamipide inhibit CCL4 induced increase in ALT and AST significantly (p < 0.05). Rebamipide exerted an antioxidant effect as it inhibits CCL4 induced increased MDA level and decreased SOD activity. Fibrotic markers assay revealed that repamipide (60 or 100 mg/kg/day) decreased the level of procol-III and HA compared to CCl4 (p < 0.05). Oral administration of Rebamipide was associated with a significant increase (p < 0.05) of PGE2 and IL-10. Rebamipide especially at the dose of (100 mg/kg/day) restores liver histology structure and abolish collagen precipitation in liver tissues. Conclusion: Rebamipide retards hepatic fibrosis induced by CCL4 may be through the induction of PGE2 level.

Keywords: fibrotic markers, hepatic fibrosis, PGE2, rebamipide

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Authors: L. Heidari, M. Jalili Ghazizade

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This paper aims to determine the best environmental and economic scenario for Municipal Solid Waste (MSW) management of the Maku city by using Life Cycle Assessment (LCA) approach. The functional elements of this study are collection, transportation, and disposal of MSW in Maku city. Waste composition and density, as two key parameters of MSW, have been determined by field sampling, and then, the other important specifications of MSW like chemical formula, thermal energy and water content were calculated. These data beside other information related to collection and disposal facilities are used as a reliable source of data to assess the environmental impacts of different waste management options, including landfills, composting, recycling and energy recovery. The environmental impact of MSW management options has been investigated in 15 different scenarios by Integrated Waste Management (IWM) software. The photochemical smog, greenhouse gases, acid gases, toxic emissions, and energy consumption of each scenario are measured. Then, the environmental indices of each scenario are specified by weighting these parameters. Economic costs of scenarios have been also compared with each other based on literature. As final result, since the organic materials make more than 80% of the waste, compost can be a suitable method. Although the major part of the remaining 20% of waste can be recycled, due to the high cost of necessary equipment, the landfill option has been suggested. Therefore, the scenario with 80% composting and 20% landfilling is selected as superior environmental and economic scenario. This study shows that, to select a scenario with practical applications, simultaneously environmental and economic aspects of different scenarios must be considered.

Keywords: IWM software, life cycle assessment, Maku, municipal solid waste management

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Authors: D. Shehu, Z. Alias

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Glutathione S-transferases (GSTs) are family of enzymes that function in the detoxification of variety of electrophilic substrates. In the present work, we report a novel zeta-like GST (designated as KKSG9) from the biphenyl/polychlorobiphenyl degrading organism Acidovorax sp. KKS102. KKSG9 possessed low sequence similarity but similar biochemical properties to zeta class GSTs. The gene for KKSG9 was cloned, purified and biochemically characterized. Functional analysis showed that the enzyme exhibits wider substrate specificity compared to most zeta class GSTs by reacting with 1-chloro-2,4-dinitrobenzene (CDNB), p-nitrobenzyl chloride (NBC), ethacrynic acid (EA), hydrogen peroxide, and cumene hydroperoxide (CuOOH). The enzyme also displayed dehalogenation function against dichloroacetate (a common substrate for zeta class GSTs) in addition to permethrin, and dieldrin. The functional role of Tyr12 was also investigated by site-directed mutagenesis. The mutant (Y12C) displayed low catalytic activity and dehalogenation function against all the substrates when compared with the wild type. Kinetic analysis using NBC and GSH as substrates showed that the mutant (Y12C) displayed a higher affinity for NBC when compared with the wild type, however, no significant change in GSH affinity was observed. These findings suggest that the presence of tyrosine residue in the motif might represent an evolutionary trend toward improving the catalytic activity of the enzyme. The enzyme as well could be useful in the bioremediation of various types of organochlorine pollutants.

Keywords: Acidovorax sp. KKS102, bioremediation, glutathione s-transferase, site-directed mutagenesis, zeta

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Authors: Eman M. Morgan, Hayam M. Lotfy, Yasmin M. Fayez, Mohamed Abdelkawy, Engy Shokry

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Two simple, selective, economic, safe, accurate, precise and environmentally friendly methods were developed and validated for the quantitative determination of valaciclovir (VAL) in the presence of its related substances R1 (acyclovir), R2 (guanine) in bulk powder and in the commercial pharmaceutical product containing the drug. Method A is a colorimetric method where VAL selectively reacts with ferric hydroxamate and the developed color was measured at 490 nm over a concentration range of 0.4-2 mg/mL with percentage recovery 100.05 ± 0.58 and correlation coefficient 0.9999. Method B is a reversed phase ultra performance liquid chromatographic technique (UPLC) which is considered superior in technology to the high-performance liquid chromatography with respect to speed, resolution, solvent consumption, time, and cost of analysis. Efficient separation was achieved on Agilent Zorbax CN column using ammonium acetate (0.1%) and acetonitrile as a mobile phase in a linear gradient program. Elution time for the separation was less than 5 min and ultraviolet detection was carried out at 256 nm over a concentration range of 2-50 μg/mL with mean percentage recovery 100.11±0.55 and correlation coefficient 0.9999. The proposed methods were fully validated as per International Conference on Harmonization specifications and effectively applied for the analysis of valaciclovir in pure form and tablets dosage form. Statistical comparison of the results obtained by the proposed and official or reported methods revealed no significant difference in the performance of these methods regarding the accuracy and precision respectively.

Keywords: hydroxamic acid, related substances, UPLC, valaciclovir

Procedia PDF Downloads 245

Authors: Ji Won Kim, Moo Yeol Lee, Keon Wook Kang

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GV-1001, 16 amino acid fragment of human telomerase reverse transcriptase catalytic subunit (hTERT), has been developed as an injectable cancer vaccine for many types of solid tumors showing high-level of telomerase activity. In the present study, we evaluated the anti-cancer effect of GV-1001 on androgen-receptor-positive prostate cancer. Two signaling pathways, Gs-adenylate cyclase-cAMP and Gq-IP3-Ca2+ pathways play a central role in GnRH receptor (GnRHR)-mediated activities. We found that leuprolide acetate (LA) mainly acted on Gq-mediated Ca2+ signaling, while GV-1001 preferentially acted on cAMP signaling; and both the effects were counteracted by cetrorelix, a GnRHR antagonist. We further tested whether GV-1001 affects tumor growth of human prostate cancer cells in vivo. Prostate tumor xenografts were established using LNCap, androgen receptor-positive prostate cancer cells, and the nude mice bearing tumors were subcutaneously injected with GV-1001 (0.01, 0.1, 1, 10 microg/kg/day) and LA (0.01 microg/kg/day) for 2 weeks. GV-1001 (1 and 10 microg/kg/day) significantly inhibited tumor growth of LNCap xenografts. Interestingly, mRNA expression of MMP2 and MMP9 was significantly suppressed by GV-1001 injection, but not by LA administration. Boyden chamber assay revealed that GV-1001 potently inhibited cell migration of LNCap. Our finding suggests that GV-1001 as a novel GnRHR ligand, has anti-proliferative and anti-migratory effects on androgen receptor-positive prostate cancer cells.

Keywords: GV-1001, GnRH, hTERT, prostate cancer

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Authors: M. Knezevic, V. Marecic, M. Ozanic, I. Kelava, M. Mihelcic, M. Santic

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Francisella tularensis is a highly infectious, gram-negative intracellular bacterium and the causative agent of tularemia. The bacterium has been isolated from more than 250 wild species, including protozoa cells. Since Francisella is very virulent and persists in the environment for years, the aim of this study was to investigate whether Acanthamoeba castellanii-grown F. novicida exhibits an alteration in the resistance to disinfectants. It has been shown by other intracellular pathogens, including Legionella pneumophila that bacteria grown in amoeba exhibit more resistance to disinfectants. However, there is no data showing Francisella viability behaviour after intracellular life cycle in A. castellani. In this study, the bacterial suspensions of A. castellanii-grown or in vitro-grown Francisella were treated with three different disinfectants, and the bacterial viability after disinfection treatment was determined by a colony-forming unit (CFU) counting method, transmission electron microscopy (TEM), fluorescence microscopy as well as the leakage of intracellular fluid. Our results have shown that didecyldimethylammonium chloride (DDAC) combined with isopropyl alcohol was the most effective in bacterial killing; all in vitro-grown and A. castellanii-grown F. novicida were killed after only 10s. Surprisingly, in comparison to in vitro-grown bacteria, A. castellanii-grown F. novicida was more sensitive to decontamination by the benzalkonium chloride combined with DDAC and formic acid and the polyhexamethylene biguanide (PHMB). We can conclude that the tested disinfectants exhibit antimicrobial activity by causing a loss of structural organization and integrity of the Francisella cell wall and membrane and the subsequent leakage of the intracellular contents. Finally, the results of this study clearly demonstrate that Francisella grown in A. castellanii had become more susceptible to many disinfectants.

Keywords: Acanthamoeba, disinfectant, Francisella, sensitivity

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