Search results for: hydrogen evolution reaction

Authors: Hadach Fatiha, Algouti Ahmed, Algouti Abdellah, Jdaba Naji, Es-Sarrar Othman, Mourabit Zahra

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This paper investigates the structure and evolution of diapirism in the Ait Ourir basin, located in the High Atlas of Marrakesh, using structural and sedimentological fieldwork integrated with field mapping. A tectonic-sedimentological study of the Mesozoic cover of the Ait Ourir basin area revealed that these units were subjected to important saccadic halokinetic activity, reflected by anticline structures associated with regional faults that created several synclinal mini-basins. However, the lack of seismic coverage in the study area makes the proposed interpretation based on extrapolations of information observed on the surface. In this work, we suggest that faults and salt activity led to the formation of different structures within the studied area. The growth of the Triassic evaporites at different stages during the Mesozoic is reflected by progressive and local unconformities, recorded as having different ages. These structures created high diapiric zones with reduced sedimentation, showing abrupt lateral thickness variations in several places where this activity was occurring; this is clearly defined within the Wanina and Jbel Sour’s mini-basins, where the Senonian was observed to rest at an angular unconformity over the entire sedimentary cover encompassing the time period from the Liassic to the Turonian. The diapirism associated with the major faults, especially encountered between the basins, is often accompanied by late Triassic volcanic material. This diapir-fault relationship resulted in shallow and often depocentric zones in a pull-apart system within a distensive context.

Keywords: diapir, evaporites, faults, pull-apart, Mesozoic cover, Ait Ourir, western High Atlas, Morocco

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Authors: Masauso Moses Phiri

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Frequent glucose monitoring is essential to the management of diabetes. Plasmonic enzyme-based glucose biosensors have the advantages of greater specificity, simplicity and rapidity. The aim of this study was to develop a rapid plasmonic colorimetric glucose biosensor based on biocatalytic enlargement of AuNS guided by GOx. Gold nanoparticles of 18 nm in diameter were synthesized using the citrate method. Using these as seeds, a modified seeded method for the synthesis of monodispersed gold nanostars was followed. Both the spherical and star-shaped nanoparticles were characterized using ultra-violet visible spectroscopy, agarose gel electrophoresis, dynamic light scattering, high-resolution transmission electron microscopy and energy-dispersive X-ray spectroscopy. The feasibility of a plasmonic colorimetric assay through growth of AuNS by silver coating in the presence of hydrogen peroxide was investigated by several control and optimization experiments. Conditions for excellent sensing such as the concentration of the detection solution in the presence of 20 µL AuNS, 10 mM of 2-(N-morpholino) ethanesulfonic acid (MES), ammonia and hydrogen peroxide were optimized. Using the optimized conditions, the glucose assay was developed by adding 5mM of GOx to the solution and varying concentrations of glucose to it. Kinetic readings, as well as color changes, were observed. The results showed that the absorbance values of the AuNS were blue shifting and increasing as the concentration of glucose was elevated. Control experiments indicated no growth of AuNS in the absence of GOx, glucose or molecular O₂. Increased glucose concentration led to an enhanced growth of AuNS. The detection of glucose was also done by naked-eye. The color development was near complete in ± 10 minutes. The kinetic readings which were monitored at 450 and 560 nm showed that the assay could discriminate between different concentrations of glucose by ± 50 seconds and near complete at ± 120 seconds. A calibration curve for the qualitative measurement of glucose was derived. The magnitude of wavelength shifts and absorbance values increased concomitantly with glucose concentrations until 90 µg/mL. Beyond that, it leveled off. The lowest amount of glucose that could produce a blue shift in the localized surface plasmon resonance (LSPR) absorption maxima was found to be 10 – 90 µg/mL. The limit of detection was 0.12 µg/mL. This enabled the construction of a direct sensitivity plasmonic colorimetric detection of glucose using AuNS that was rapid, sensitive and cost-effective with naked-eye detection. It has great potential for transfer of technology for point-of-care devices.

Keywords: colorimetric, gold nanostars, glucose, glucose oxidase, plasmonic

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Authors: Hager Elsheikh, Matthias Sendler, Juliana Glaubnitz

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In pancreatitis, an inflammatory reaction occurs in the pancreatic secretory cells due to premature activation of proteases, leading to pancreatic self-digestion and necrotic cell death of acinar cells. Acute pancreatitis in patients is characterized by a severe immune reaction that could lead to serious complications, such as organ failure or septic shock, if left untreated. Chronic pancreatitis is a recurrence of episodes of acute pancreatitis resulting in a fibro-inflammatory immune response, in which the type 2 immune response is primarily driven by AAMs in the pancreas. One of the most important signaling pathways for M2 macrophage activation is the IL-4/STAT6 pathway. Pancreatic fibrosis is induced by the hyperactivation of pancreatic stellate cells by dysregulation in the inflammatory response, leading to further damage, autodigestion and possibly necrosis of pancreatic acinar cells. The aim of this research is to investigate the effect of STAT6 knockout in disease severity and development of fibrosis wound healing in the presence of different macrophage populations, regulated by the type 2 immune response, after inducing chronic and/or acute pancreatitis in mice models via cerulean injection. We further investigate the influence of the JAK/STAT6 signaling pathway on the balance of fibrosis and regeneration in STAT6 deficient and wild-type mice. The characterization of resident and recruited macrophages will provide insight into the influence of the JAK/STAT6 signaling pathway on infiltrating cells and, ultimately, tissue fibrosis and disease severity.

Keywords: acute and chronic pancreatitis, tissue regeneration, macrophage polarization, Gastroenterology

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Authors: Hellal Abdelkader, Chafaa Salah, Touafri Lasnouni

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A simple, efficient and general method has been developed for the high diastereoselective synthesis of diethyl α-aminophosphonates in water through “one-pot” three-component reaction of aromatic aldehydes, aminophenols and dialkyl phosphites in the presence of a low catalytic amount (10mol%) of phosphorous acid as highly stable catalyst is described.

Keywords: DFT, HOMO-LUMO, phosphonic acid, aminophenols

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Authors: Md. Hafezur Rahaman, Mohd. Maniruzzaman, Md. Shadiqul Islam, Md. Masud Rana

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With the growth of environmental awareness, enormous researches are running to develop the next generation materials based on sustainability, eco-competence, and green chemistry to preserve and protect the environment. Due to biodegradability and biocompatibility, poly (L-lactic acid) (PLLA) has a great interest in ecological and medical applications. Also, cellulose is one of the most abundant biodegradable, renewable polymers found in nature. It has several advantages such as low cost, high mechanical strength, biodegradability and so on. Recently, an immense deal of attention has been paid for the scientific and technological development of α-cellulose based composite material. PLLA could be used for grafting of cellulose to improve the compatibility prior to the composite preparation. Here it is quite difficult to form a bond between lower hydrophilic molecules like PLLA and α-cellulose. Dimmers and oligomers can easily be grafted onto the surface of the cellulose by ring opening or polycondensation method due to their low molecular weight. In this research, α-cellulose extracted from jute fiber is grafted with oligo(D-lactic acid) (ODLA) via graft polycondensation reaction in presence of para-toluene sulphonic acid and potassium persulphate in toluene at 130°C for 9 hours under 380 mmHg. Here ODLA is synthesized by ring opening polymerization of D-lactides in the presence of stannous octoate (0.03 wt% of lactide) and D-lactic acids at 140°C for 10 hours. Composites of PLLA with ODLA grafted α-cellulose are prepared by solution mixing and film casting method. Confirmation of grafting was carried out through FTIR spectroscopy and SEM analysis. A strongest carbonyl peak of FTIR spectroscopy at 1728 cm⁻¹ of ODLA grafted α-cellulose confirms the grafting of ODLA onto α-cellulose which is absent in α-cellulose. It is also observed from SEM photographs that there are some white areas (spot) on ODLA grafted α-cellulose as compared to α-cellulose may indicate the grafting of ODLA and consistent with FTIR results. Analysis of the composites is carried out by FTIR, SEM, WAXD and thermal gravimetric analyzer. Most of the FTIR characteristic absorption peak of the composites shifted to higher wave number with increasing peak area may provide a confirmation that PLLA and grafted cellulose have better compatibility in composites via intermolecular hydrogen bonding and this supports previously published results. Grafted α-cellulose distributions in composites are uniform which is observed by SEM analysis. WAXD studied show that only homo-crystalline structures of PLLA present in the composites. Thermal stability of the composites is enhanced with increasing the percentages of ODLA grafted α-cellulose in composites. As a consequence, the resultant composites have a resistance toward the thermal degradation. The effects of length of the grafted chain and biodegradability of the composites will be studied in further research.

Keywords: α-cellulose, composite, graft polycondensation, oligo(D-lactic acid), poly(L-lactic acid)

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Authors: Kiran P. Chadayamuri, Saransh Bagdi

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Increasing imbalance between energy supply and demand has made nations and companies involved in the energy sector to boost up their research and find suitable solutions. With the high rates at which conventional oil and gas resources are depleting, efficient exploration and exploitation of heavy oil could just be the answer. Heavy oil may be defined as crude oil having API gravity value of less than 20⁰. They are highly viscous, have low hydrogen to carbon ratios and are known to produce high carbon residues. They have high contents of asphaltenes, heavy metals, sulphur and nitrogen in them. Due to these properties extraction, transportation and refining of crude oil have its share of challenges. Lack of suitable technology has hindered its production in the past, but now things are going in a more positive direction. The aim of this paper is to study the various advantages of heavy oil, associated limitations and its feasibility as a fuel of the future.

Keywords: energy, heavy oil, fuel, future

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Authors: Bummo Ahn

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Palpation is widely used to measure soft tissue firmness or stiffness in the living condition in order to apply detection, diagnosis, and treatment of tumors, scar tissue, abnormal muscle tone, or muscle spasticity. Since these methods are subjective and depend on the proficiency level, it is concluded that there are other diagnoses depending on the condition of the experts and the results are not objective. The mechanical property obtained by using the elasticity of the tissue is important to calculate a predictive variable for monitoring abnormal tissues. If the mechanical load such as reaction force on the foot increases in the same region under the same conditions, the mechanical property of the tissue is changed. Therefore, objective diagnosis is possible not only for experts but also for patients using this quantitative information. Furthermore, the portable system also allows non-experts to easily diagnose at home, not in hospitals or institutions. In this paper, we introduce a portable palpation system that can be used to measure the mechanical properties of human tissue, which can be applied to monitor diabetic foot ulceration patients with measuring the mechanical property change of foot tissue. The system was designed to be smaller and portable in comparison with the conventional palpation systems. It is consists of the probe, the force sensor, linear actuator, micro control unit, the display module, battery, and housing. Using this system, we performed validation experiments by applying different palpations (3 and 5 mm) to soft tissue (silicone rubber) and measured reaction forces. In addition, we estimated the elastic moduli of the soft tissue against different palpations and compare the estimated elastic moduli that show similar value even if the palpation depths are different.

Keywords: palpation probe, portable, diabetic foot ulceration, monitoring, mechanical property

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Authors: M. Seguini, D. Nedjar

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An accuracy nonlinear analysis of a deep beam resting on elastic perfectly plastic soil is carried out in this study. In fact, a nonlinear finite element modeling for large deflection and moderate rotation of Euler-Bernoulli beam resting on linear and nonlinear random soil is investigated. The geometric nonlinear analysis of the beam is based on the theory of von Kàrmàn, where the Newton-Raphson incremental iteration method is implemented in a Matlab code to solve the nonlinear equation of the soil-beam interaction system. However, two analyses (deterministic and probabilistic) are proposed to verify the accuracy and the efficiency of the proposed model where the theory of the local average based on the Monte Carlo approach is used to analyze the effect of the spatial variability of the soil properties on the nonlinear beam response. The effect of six main parameters are investigated: the external load, the length of a beam, the coefficient of subgrade reaction of the soil, the Young’s modulus of the beam, the coefficient of variation and the correlation length of the soil’s coefficient of subgrade reaction. A comparison between the beam resting on linear and nonlinear soil models is presented for different beam’s length and external load. Numerical results have been obtained for the combination of the geometric nonlinearity of beam and material nonlinearity of random soil. This comparison highlighted the need of including the material nonlinearity and spatial variability of the soil in the geometric nonlinear analysis, when the beam undergoes large deflections.

Keywords: finite element method, geometric nonlinearity, material nonlinearity, soil-structure interaction, spatial variability

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Authors: Dian Muzdalifah, Anastasia F. Devi, Zatil A. Athaillah, Linar Z. Udin

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Tempe is a functional food prepared from soybeans through Rhizopus spp fermentation. It is well known as functional food, originated from Indonesia. Most studies on tempe functionalities refer to ripe (48 h fermentation) tempe and only limited studies discuss overripe tempe while longer fermentation time possibly increased tempe health benefit. Hence, the present study was performed to investigate the cytotoxic activity againts MCF-7 breast cancer cells and antioxidant property of tempe prepared from 0–156 h of fermentation. Tempe samples were dried and extracted with acetone and ethanol, respectively. Their extracts were used for subsequent analysis. The cytotoxic activity was assessed on MCF 7 breast cancer cells using Alamar Blue method. The antioxidant activity was determined by DPPH free radical scavenging assay. The results indicated that acetone extracts of 108 h tempe had a potent cytotoxic activity against MCF-7 breast cancer cells (IC50 = 2.54 ± 0,30 μg/mL). Ethanol extracts of 108 h tempe also showed the potency, but at slightly higher IC50 (5.20 ± 1.01 μg/mL). Both acetone and ethanol extracts of 108 and 120 h tempe showed high antioxidant activity expressed as percent inhibition with no significant difference. However, acetone extracts of 120 h tempe (81.31 ± 3.70 %) had better ability to inhibit oxidation reaction than that of ethanol extracts (75.77 ± 6.00 %). It can be concluded that the cytotoxic activity of tempe from 0–156 h of fermentation is positively correlated to their corresponding antioxidant property. Longer fermentation time, up to 108 h, increased the ability of tempe to inhibit the growth of MCF-7 breast cancer cells and oxidative reaction. But extended fermentation time, up to 156 h, tends to decrease its ability. Further studies are encouraged to identify the active components contained in each extract.

Keywords: antioxidant property, cytotoxic activity, extracts, overripe tempeh

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Authors: Víctor D. Rodríguez, Edith P. Estupiñán, Juan C. Martínez

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The evolution in mobile broadband technologies has allowed to increase the download rates in users considering the current services. The evaluation of technical parameters at the link level is of vital importance to validate the quality and veracity of the connection, thus avoiding large losses of data, time and productivity. Some of these failures may occur between the eNodeB (Evolved Node B) and the user equipment (UE), so the link between the end device and the base station can be observed. LTE (Long Term Evolution) is considered one of the IP-oriented mobile broadband technologies that work stably for data and VoIP (Voice Over IP) for those devices that have that feature. This research presents a technical analysis of the connection and channeling processes between UE and eNodeB with the TAC (Tracking Area Code) variables, and analysis of performance variables (Throughput, Signal to Interference and Noise Ratio (SINR)). Three measurement scenarios were proposed in the city of Bogotá using QualiPoc, where two operators were evaluated (Operator 1 and Operator 2). Once the data were obtained, an analysis of the variables was performed determining that the data obtained in transmission modes vary depending on the parameters BLER (Block Error Rate), performance and SNR (Signal-to-Noise Ratio). In the case of both operators, differences in transmission modes are detected and this is reflected in the quality of the signal. In addition, due to the fact that both operators work in different frequencies, it can be seen that Operator 1, despite having spectrum in Band 7 (2600 MHz), together with Operator 2, is reassigning to another frequency, a lower band, which is AWS (1700 MHz), but the difference in signal quality with respect to the establishment with data by the provider Operator 2 and the difference found in the transmission modes determined by the eNodeB in Operator 1 is remarkable.

Keywords: BLER, LTE, network, qualipoc, SNR.

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Authors: Shiliang Wu, Huanxin Zhang, Aditya Kumar

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There have been increasing concerns on atmospheric mercury as a toxic and bioaccumulative pollutant in the global environment. Global change, including changes in climate change, land use, land cover and wildfires activities can all have significant impacts on atmospheric mercury. In this study, we use a global chemical transport model (GEOS-Chem) to examine the potential impacts from global change on atmospheric mercury. All of these factors in the context of global change are found to have significant impacts on the long-term evolution of atmospheric mercury and can substantially alter the global source-receptor relationships for mercury. We also estimate the global Hg emissions from wildfires for present-day and the potential impacts from the 2000-2050 changes in climate, land use and land cover and Hg anthropogenic emissions by combining statistical analysis with global data on vegetation type and coverage as well as fire activities. Present global Hg wildfire emissions are estimated to be 612 Mg year-1. Africa is the dominant source region (43.8% of global emissions), followed by Eurasia (31%) and South America (16.6%). We find significant perturbations to wildfire emissions of Hg in the context of global change, driven by the projected changes in climate, land use and land cover and Hg anthropogenic emissions. 2000-2050 climate change could increase Hg emissions by 14% globally. Projected changes in land use by 2050 could decrease the global Hg emissions from wildfires by 13% mainly driven by a decline in African emissions due to significant agricultural land expansion. Future land cover changes could lead to significant increases in Hg emissions over some regions (+32% North America, +14% Africa, +13% Eurasia). Potential enrichment of terrestrial ecosystems in 2050 in response to changes in Hg anthropogenic emissions could increase Hg wildfire emissions both globally (+28%) and regionally. Our results indicate that the future evolution of climate, land use and land cover and Hg anthropogenic emissions are all important factors affecting Hg wildfire emissions in the coming decades.

Keywords: climate change, land use, land cover, wildfires

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Authors: Lise Paesen, Marielle Leijten

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People diagnosed with probable Alzheimer disease suffer from an impairment of their language capacities; a gradual impairment which affects both their spoken and written communication. Our study aims at characterising the language decline in DAT patients with the use of congeneric elicited writing tasks. Within these tasks, a descriptive text has to be written based upon images with which the participants are confronted. A randomised set of images allows us to present the participants with a different task on every encounter, thus allowing us to avoid a recognition effect in this iterative study. This method is a revision from previous studies, in which participants were presented with a larger picture depicting an entire scene. In order to create the randomised set of images, existing pictures were adapted following strict criteria (e.g. frequency, AoA, colour, ...). The resulting data set contained 50 images, belonging to several categories (vehicles, animals, humans, and objects). A pre-test was constructed to validate the created picture set; most images had been used before in spoken picture naming tasks. Hence the same reaction times ought to be triggered in the typed picture naming task. Once validated, the effectiveness of the descriptive tasks was assessed. First, the participants (n=60 students, n=40 healthy elderly) performed a typing task, which provided information about the typing speed of each individual. Secondly, two descriptive writing tasks were carried out, one simple and one complex. The simple task contains 4 images (1 animal, 2 objects, 1 vehicle) and only contains elements with high frequency, a young AoA (<6 years), and fast reaction times. Slow reaction times, a later AoA (≥ 6 years) and low frequency were criteria for the complex task. This task uses 6 images (2 animals, 1 human, 2 objects and 1 vehicle). The data were collected with the keystroke logging programme Inputlog. Keystroke logging tools log and time stamp keystroke activity to reconstruct and describe text production processes. The data were analysed using a selection of writing process and product variables, such as general writing process measures, detailed pause analysis, linguistic analysis, and text length. As a covariate, the intrapersonal interkey transition times from the typing task were taken into account. The pre-test indicated that the new images lead to similar or even faster reaction times compared to the original images. All the images were therefore used in the main study. The produced texts of the description tasks were significantly longer compared to previous studies, providing sufficient text and process data for analyses. Preliminary analysis shows that the amount of words produced differed significantly between the healthy elderly and the students, as did the mean length of production bursts, even though both groups needed the same time to produce their texts. However, the elderly took significantly more time to produce the complex task than the simple task. Nevertheless, the amount of words per minute remained comparable between simple and complex. The pauses within and before words varied, even when taking personal typing abilities (obtained by the typing task) into account.

Keywords: Alzheimer's disease, experimental design, language decline, writing process

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Authors: Lemar Blake, Chris Oura, Ayanna C. N. Phillips Savage

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Improved DNA sequencing technology has greatly enhanced bacterial identification, especially for organisms that are difficult to culture. Mycobacteriosis with consistent hyphema, bilateral exophthalmia, open mouth gape and ocular lesions, were observed in various fish populations at the School of Veterinary Medicine, Aquaculture/Aquatic Animal Health Unit. Objective: To identify the species of Mycobacterium that is affecting aquarium fish at the School of Veterinary Medicine, Aquaculture/Aquatic Animal Health Unit. Method: A total of 13 fish samples were collected and analyzed via: Ziehl-Neelsen, conventional polymerase chain reaction (PCR) and real-time PCR. These tests were carried out simultaneously for confirmation. The following combination of conventional primers: 16s rRNA (564 bp), rpoB (396 bp), sod (408 bp) were used. Concatenation of the gene fragments was carried out to phylogenetically classify the organism. Results: Acid fast non-branching bacilli were detected in all samples from homogenized internal organs. All 13 acid fast samples were positive for Mycobacterium via real-time PCR. Partial gene sequences using all three primer sets were obtained from two samples and demonstrated a novel strain. A strain 99% related to Mycobacterium marinum was also confirmed in one sample, using 16srRNA and rpoB genes. The two novel strains were clustered with the rapid growers and strains that are known to affect humans. Conclusions: Phylogenetic analysis demonstrated two novel Mycobacterium strains with the potential of being zoonotic and one strain 99% related to Mycobacterium marinum.

Keywords: polymerase chain reaction, phylogenetic, DNA sequencing, zoonotic

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Authors: Sujata Sharma, Avinash Singh, Lovely Gautam, Pradeep Sharma, Mau Sinha, Asha Bhushan, Punit Kaur, Tej P. Singh

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Peptidyl-tRNA hydrolase (Pth) Pth is an essential bacterial enzyme that catalyzes the release of free tRNA and peptide moeities from peptidyl tRNAs during stalling of protein synthesis. In order to design inhibitors of Pth from Pseudomonas aeruginosa (PaPth), we have determined the structures of PaPth in its native state and in the bound states with two compounds, amino acylate-tRNA analogue (AAtA) and 5-azacytidine (AZAC). The peptidyl-tRNA hydrolase gene from Pseudomonas aeruginosa was amplified by Phusion High-Fidelity DNA Polymerase using forward and reverse primers, respectively. The E. coliBL21 (λDE3) strain was used for expression of the recombinant peptidyl-tRNA hydrolase from Pseudomonas aeruginosa. The protein was purified using a Ni-NTA superflow column. The crystallization experiments were carried out using hanging drop vapour diffusion method. The crystals diffracted to 1.50 Å resolution. The data were processed using HKL-2000. The polypeptide chain of PaPth consists of 194 amino acid residues from Met1 to Ala194. The centrally located β-structure is surrounded by α-helices from all sides except the side that has entrance to the substrate binding site. The structures of the complexes of PaPth with AAtA and AZAC showed the ligands bound to PaPth in the substrate binding cleft and interacted with protein atoms extensively. The residues that formed intermolecular hydrogen bonds with the atoms of AAtA included Asn12, His22, Asn70, Gly113, Asn116, Ser148, and Glu161 of the symmetry related molecule. The amino acids that were involved in hydrogen bonded interactions in case of AZAC included, His22, Gly113, Asn116, and Ser148. As indicated by fittings of two ligands and the number of interactions made by them with protein atoms, AAtA appears to be a more compatible with the structure of the substrate binding cleft. However, there is a further scope to achieve a better stacking than that of O-tyrosyl moiety because it is not still ideally stacked. These observations about the interactions between the protein and ligands have provided the information about the mode of binding of ligands, nature and number of interactions. This information may be useful for the design of tight inhibitors of Pth enzymes.

Keywords: peptidyl tRNA hydrolase, Acinetobacter baumannii, Pth enzymes, O-tyrosyl

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Authors: B. Kulaga, A. Cinti, F. J. Mazzocchini

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Despite the fact that growing academic attention on dark tourism is a fairly recent phenomenon, among the various reasons for travelling death-related ones, are very ancient. Furthermore, the darker side of human nature has always been fascinated and curious regarding death, or at least, man has always tried to learn lessons from death. This study proposes to describe the phenomenon of dark tourism related to the 2016 earthquake in Central Italy, deadly for 302 people and highly destructive for the rural areas of Lazio, Marche, and Umbria Regions. The primary objective is to examine the motivation-experience relationship in a dark tourism site, using the structural equation model, applied for the first time to a dark tourism research in 2016, in a study conducted after the Beichuan earthquake. The findings of the current study are derived from the calculations conducted on primary data compiled from 350 tourists in the areas mostly affected by the 2016 earthquake, including the town of Amatrice, near the epicenter, Castelluccio, Norcia, Ussita and Visso, through conducting a Likert scale survey. Furthermore, we use the structural equation model to examine the motivation behind dark travel and how this experience can influence the motivation and emotional reaction of tourists. Expected findings are in line with the previous study mentioned above, indicating that: not all tourists visit the thanatourism sites for dark tourism purpose, tourists’ emotional reactions influence more heavily the emotional tourist experience than cognitive experiences do, and curious visitors are likely to engage cognitively by learning about the incident or related issues.

Keywords: dark tourism, emotional reaction, experience, motivation, structural equation model

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Authors: Abbas Pourreza

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MicroRNAs (miRNAs) are small single-stranded non-coding RNAs. They are almost 18-25 nucleotides long and very conservative through evolution. They are involved in adjusting the expression of numerous genes due to the existence of a complementary region, generally in the 3' untranslated regions (UTR) of target genes, against particular mRNAs in the cell. Also, miRNAs have been proven to be involved in cell development, differentiation, proliferation, and apoptosis. More than 2000 miRNAs have been recognized in human cells, and these miRNAs adjust approximately one-third of all genes in human cells. Dysregulation of miRNA originated from abnormal DNA methylation patterns of the locus, cause to down-regulated or overexpression of miRNAs, and it may affect tumor formation or development of it. Breast cancer (BC) is the most commonly identified cancer, the most prevalent cancer (23%), and the second-leading (14%) mortality in all types of cancer in females. BC can be classified based on the status (+/−) of the hormone receptors, including estrogen receptor (ER), progesterone receptor (PR), and the Receptor tyrosine-protein kinase erbB-2 (ERBB2 or HER2). Currently, there are four main molecular subtypes of BC: luminal A, approximately 50–60 % of BCs; luminal B, 10–20 %; HER2 positive, 15–20 %, and 10–20 % considered Basal (triple-negative breast cancer (TNBC)) subtype. Aberrant expression of miR-145, miR-21, miR-10b, miR-125a, and miR-206 was detected by Stem-loop real-time RT-PCR in BC cases. Breast tumor formation and development may result from down-regulation of a tumor suppressor miRNA such as miR-145, miR-125a, and miR-206 and/or overexpression of an oncogenic miRNA such as miR-21 and miR-10b. MiR-125a, miR-206, miR-145, miR-21, and miR-10b are hugely predicted to be new tumor markers for the diagnosis and prognosis of BC. MiR-21 and miR-125a could play a part in the treatment of HER-2-positive breast cancer cells, while miR-145 and miR-206 could speed up the evolution of cure techniques for TNBC. To conclude, miRNAs will be presented as hopeful molecules to be used in the primary diagnosis, prognosis, and treatment of BC and battle as opposed to its developed drug resistance.

Keywords: breast cancer, HER2 positive, miRNA, TNBC

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Authors: Guillaume Bélanger, Jean-Philippe Fontaine, Clémence Hauduc

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There is an undeniable thirst from organic chemists and from the pharmaceutical industry to access complex alkaloids with short syntheses. While medicinal chemists are interested in the fascinating wide range of biological properties of alkaloids, synthetic chemists are rather interested in finding new routes to access these challenging natural products of often low availability from nature. To synthesize complex polycyclic cores of natural products, reaction cascades or sequences performed one-pot offer a neat advantage over classical methods for their rapid increase in molecular complexity in a single operation. In counterpart, reaction cascades need to be run on substrates bearing all the required functional groups necessary for the key cyclizations. Chemoselectivity is thus a major issue associated with such a strategy, in addition to diastereocontrol and regiocontrol for the overall transformation. In the pursuit of synthetic efficiency, our research group developed an innovative one-pot transformation of linear substrates into bi- and tricyclic adducts applied to the construction of Aspidospermatan-type alkaloids. The latter is a rich class of indole alkaloids bearing a unique bridged azatricyclic core. Despite many efforts toward the synthesis of members of this family, efficient and versatile synthetic routes are still coveted. Indeed, very short, non-racemic approaches are rather scarce: for example, in the cases of aspidospermidine and aspidospermine, syntheses are all fifteen steps and over. We envisaged a unified approach to access several members of the Aspidospermatan alkaloids family. The key sequence features a highly chemoselective formamide activation that triggers a Vilsmeier-Haack cyclization, followed by an azomethine ylide generation and intramolecular cycloaddition. Despite the high density and variety of functional groups on the substrates (electron-rich and electron-poor alkenes, nitrile, amide, ester, enol ether), the sequence generated three new carbon-carbon bonds and three rings in a single operation with good yield and high chemoselectivity. A detailed study of amide, nucleophile, and dipolarophile variations to finally get to the successful combination required for the key transformation will be presented. To complete the indoline fragment of the natural products, we developed an original approach. Indeed, all reported routes to Aspidospermatan alkaloids introduce the indoline or indole early in the synthesis. In our work, the indoline needs to be installed on the azatricyclic core after the key cyclization sequence. As a result, typical Fischer indolization is not suited since this reaction is known to fail on such substrates. We thus envisaged a unique photocatalyzed decarboxylative radical cyclization. The development of this reaction as well as the scope and limitations of the methodology, will also be presented. The original Vilsmeier-Haack and azomethine ylide cyclization sequence as well as the new photocatalyzed decarboxylative radical cyclization will undoubtedly open access to new routes toward polycyclic indole alkaloids and derivatives of pharmaceutical interest in general.

Keywords: Aspidospermatan alkaloids, azomethine ylide cycloaddition, decarboxylative radical cyclization, indole and indoline synthesis, one-pot sequential cyclizations, photocatalysis, Vilsmeier-Haack Cyclization

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Authors: Mei Han, Jinkou Zhao, Yuan Yao, Liangui Feng, Xianbin Ding, Guohui Wu, Chao Zhou, Lin Ouyang, Rongrong Lu, Bo Zhang

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To compare the HIV incidence estimated using BED capture enzyme linked immunosorbent assay (BED-CEIA) and observed in a cohort against the HIV incidence among men who have sex with men (MSM) measured by pooling polymerase chain reaction (pooling-PCR). A total of 617 MSM subjects were included in a respondent driven sampling survey in Chongqing in 2008. Among the 129 that were tested HIV antibody positive, 102 were defined with long-term infection, 27 were assessed for recent HIV infection (RHI) using BED-CEIA. The remaining 488 HIV negative subjects were enrolled to the prospective cohort and followed-up every 6 months to monitor HIV seroconversion. All of the 488 HIV negative specimens were assessed for acute HIV infection (AHI) using pooling-PCR. Among the 488 negative subjects in the open cohort, 214 (43.9%) were followed-up for six months, with 107 person-years of observation and 14 subjects seroconverted. The observed HIV incidence was 12.5 per 100 person-years (95% CI=9.1-15.7). Among the 488 HIV negative specimens, 5 were identified with acute HIV infection using pooling-PCR at an annual rate of 14.02% (95% CI=1.73-26.30). The estimated HIV-1 incidence was 12.02% (95% CI=7.49-16.56) based on BED-CEIA. The HIV incidence estimated with three different approaches was different among subgroups. In the highly HIV prevalent MSM, it costs US$ 1724 to detect one AHI case, while detection of one case of RHI with BED assay costs only US$ 42. Three approaches generated comparable and high HIV incidences, pooling PCR and prospective cohort are more close to the true level of incidence, while BED-CEIA seemed to be the most convenient and economical approach for at-risk population’s HIV incidence evaluation at the beginning of HIV pandemic. HIV-1 incidences were alarmingly high among MSM population in Chongqing, particularly within the subgroup under 25 years of age and those migrants aged between 25 to 34 years.

Keywords: BED-CEIA, HIV, incidence, pooled PCR, prospective cohort

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Authors: M. Zielina, W. Dabrowski, E. Radziszewska-Zielina

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Several different cements have been tested to evaluate their potential to leach calcium, chromium and aluminum ions in soft water environment. The research allows comparing some different cements in order to the potential risk of water contamination. This can be done only in the same environment. To reach the results in reasonable short time intervals and to make heavy metals measurements with high accuracy, demineralized water was used. In this case the conditions of experiments are far away from the water supply practice, but short time experiments and measurably high concentrations of elements in the water solution are an important advantage. Moreover leaching mechanisms can be recognized, our experiments reported here refer to this kind of cements evaluation.

Keywords: concrete corrosion, hydrogen sulfide, odors, reinforced concrete sewers, sewerage

Procedia PDF Downloads 190

Authors: Yuri A. Kalvachev, Totka D. Todorova

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Zeolite mordenite has been investigated as a transition metal support for the preparation of efficient catalysts in the oxidation of volatile organic compounds (VOCs). The highly crystalline mordenite samples were treated with hydrofluoric acid and ammonium fluoride to get hierarchical material with secondary porosity. The obtained supports by this method have a high active surface area, good diffusion properties and prevent the extraction of metal components during catalytic reactions. The active metal phases platinum and copper were loaded by impregnation on both mordenite materials (parent and acid treated counterparts). Monometalic Pt and Cu, and bimetallic Pt/Cu catalysts were obtained. The metal phases were fine dispersed as nanoparticles on the functional porous materials. The catalysts synthesized in this way were investigated in the reaction of complete oxidation of propane and benzene. Platinum, copper and platinum/copper were loaded and there catalytic activity was investigated and compared. All samples are characterized by X-ray diffraction analysis, nitrogen adsorption, scanning electron microscopy (SEM), X-ray photoelectron measurements (XPS) and temperature programed reduction (TPR). The catalytic activity of the samples obtained is investigated in the reaction of complete oxidation of propane and benzene by using of Gas Chromatography (GC). The oxidation of three organic molecules was investigated—methane, propane and benzene. The activity of metal loaded mordenite catalysts for methane oxidation is almost the same for parent and treated mordenite as a support. For bigger molecules as propane and benzene, the activity of catalysts based on treated mordenite is higher than those based on parent zeolite.

Keywords: metal loaded catalysts, mordenite, VOCs oxidation, zeolites

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Authors: Abobakr Khidir Ziyada, Cecilia Devi Wilfred

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In the present work, a new series of 3-propanenitrile imidazolium-based Room Temperature Ionic Liquids (RTILs), incorporating dioctyl sulfosuccinate (DOSS) were prepared by reacting imidazole with acrylonitrile and then reacting the product with allyl chloride, 2-chloroethanol, and benzyl chloride. After the reaction had been completed, metathesis reaction was carried out using sodium dioctyl sulfosuccinate. The densities and viscosities of the present RTILs were measured at atmospheric pressure at T=293.15 to 353.15 K, the refractive index was measured at T=293.15 to 333.15 K, whereas, the start and decomposition temperatures were determined at heating rate 10°C. min^-1. The thermal expansion coefficient, densities at a range of temperatures and pressures, molecular volume, molar refraction, standard entropy and the lattice energy of these RTILs were also estimated. The present RTILs showed higher densities, similar refractive indices, and higher viscosities compared to the other 1-alkyl-3-propanenitrile imidazolium-based RTILs. The densities of the present synthesized RTILs are lower compared to the other nitrile-functionalized ILs. These present RTILs showed a weak temperature dependence on the thermal expansion coefficients, αp=5.0 × 10^−4 to 7.50 × 10−4 K^-1. Empirical correlations were proposed to represent the present data on the physical properties. The lattice energy for the present RTILs was similar to other nitrile–based imidazolium RTILs. The present RTILs showed very high molar refraction when compared similar RTILs incorporating other anions.

Keywords: dioctyl sulfosuccinate, nitrile ILs, 3-propanenitrile, anion, room temperature ionic liquids, RTIL

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Authors: A. E. Emam, M. Abd Elghany

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The change in orbit evolution between collocated satellites (X, Y) inside +/-0.09 ° E/W and +/- 0.07 ° N/S cluster, after one of these satellites is placed in an inclined orbit (satellite X) and the effect of this change in the collocation safety inside the cluster window has been studied and evaluated. Several collocation scenarios had been studied in order to adjust the location of both satellites inside their cluster to maximize the separation between them and safe the mission.

Keywords: satellite, GEO, collocation, risk assessment

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Authors: Shivam Yadav, Neelam Atri

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Cyanobacteria are photoautotrophic prokaryotes able to grow in diverse ecological habitats, originated 2.5 - 3.5 billion years ago and brought oxygenic photosynthesis. Since then superoxide dismutases (SODs) acquired great significance due to their ability to catalyze detoxification of byproducts of oxygenic photosynthesis, i.e. superoxide radicals. Sequence information from several cyanobacterial genomes offers a unique opportunity to conduct a comprehensive comparative analysis of the superoxide dismutases family. In the present study, we extracted information regarding SODs from species of sequenced cyanobacteria and investigated their diversity, conservation, domain structure, and evolution. 144 putative SOD homologues were identified. SODs are present in all cyanobacterial species reflecting their significant role in survival. However, their distribution varies, fewer in unicellular marine strains whereas abundant in filamentous nitrogen-fixing cyanobacteria. Motifs and invariant amino acids typical in eukaryotic SODs were conserved well in these proteins. These SODs were classified into three major families according to their domain structures. Interestingly, they lack additional domains as found in proteins of other family. Phylogenetic relationships correspond well with phylogenies based on 16S rRNA and clustering occurs on the basis of structural characteristics such as domain organization. Similar conserved motifs and amino acids indicate that cyanobacterial SODs make use of a similar catalytic mechanism as eukaryotic SODs. Gene gain-and-loss is insignificant during SOD evolution as evidenced by absence of additional domain. This study has not only examined an overall background of sequence-structure-function interactions for the SOD gene family but also revealed variation among SOD distribution based on ecophysiological and morphological characters.

Keywords: comparative genomics, cyanobacteria, phylogeny, superoxide dismutases

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Authors: T. Rajamäki

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In clean room environment molecular contamination in very small concentrations can cause significant harm for the components and processes. This is commonly referred as airborne molecular contamination (AMC). There is a shortage of high sensitivity continuous measurement data for existence and behavior of several of these contaminants. Accordingly, in most cases correlation between concentration of harmful molecules and their effect on processes is not known. In addition, the formation and distribution of contaminating molecules are unclear. In this work sensitive optical techniques are applied in clean room facilities for investigation of concentrations, forming mechanisms and effects of contaminating molecules. Special emphasis is on reactive acid and base gases ammonia (NH3) and hydrogen fluoride (HF). They are the key chemicals in several operations taking place in clean room processes.

Keywords: AMC, clean room, concentration, reactive gas

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Authors: Taraneh Estedlal

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The objective of this study was to review in-vivo and in-vitro studies to compare the effectiveness of bulk-fill and conventional resin composites with regard to marginal adaptation, polymerization shrinkage, and other mechanical properties.PubMed and Scopus databases was investigated for in-vitro studies and randomized clinical trials comparing incidence of fractures, color stability, marginal adaptation, pain and discomfort, recurrent caries, occlusion, pulpal reaction, and proper proximal contacts of restorations made with conventional and bulk resins. The failure rate of conventional and flowable bulk-fill resin composites was not significantly different to sculptable bulk-fill resin composites. The objective of this study was to review in-vivo and in-vitro studies to compare the effectiveness of bulk-fill and conventional resin composites with regard to marginal adaptation, polymerization shrinkage, and other mechanical properties. PubMed and Scopus databases was investigated for in-vitro studies and randomized clinical trials comparing one of the pearlier mentioned properties between bulk-fill and control composites. Despite differences in physical and in-vitro properties, failure rate of conventional and flowable bulk-fill resin composites was not significantly different to sculptable bulk-fill resin composites.

Keywords: polymerization shrinkage, color stability, marginal adaptation, recurrent caries, occlusion, pulpal reaction

Procedia PDF Downloads 134

Authors: Reza Miresmaeili, Asghar Heydari Astaraee, Fereshteh Dolati

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In the present study, an analytical model based on dislocation density model was developed to simulate grain refinement in surface mechanical attrition treatment (SMAT). The correlation between SMAT time and development in plastic strain on one hand, and dislocation density evolution, on the other hand, was established to simulate the grain refinement in SMAT. A dislocation density-based constitutive material law was implemented using VUHARD subroutine. A random sequence of shots is taken into consideration for multiple impacts model using Python programming language by utilizing a random function. The simulation technique was to model each impact in a separate run and then transferring the results of each run as initial conditions for the next run (impact). The developed Finite Element (FE) model of multiple impacts describes the coverage evolution in SMAT. Simulations were run to coverage levels as high as 4500%. It is shown that the coverage implemented in the FE model is equal to the experimental coverage. It is depicted that numerical SMAT coverage parameter is adequately conforming to the well-known Avrami model. Comparison between numerical results and experimental measurements for residual stresses and depth of deformation layers confirms the performance of the established FE model for surface engineering evaluations in SMA treatment. X-ray diffraction (XRD) studies of grain refinement, including resultant grain size and dislocation density, were conducted to validate the established model. The full width at half-maximum in XRD profiles can be used to measure the grain size. Numerical results and experimental measurements of grain refinement illustrate good agreement and show the capability of established FE model to predict the gradient microstructure in SMA treatment.

Keywords: dislocation density, grain refinement, severe plastic deformation, simulation, surface mechanical attrition treatment

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Authors: Lucie Speyer, Vincent Lecocq, Séverine Humbert, Antoine Hugon

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Mesoporous alumina is commonly used as a catalyst support for the hydrotreating of heavy petroleum cuts. The process of fabrication usually involves: the synthesis of the boehmite AlOOH precursor, a kneading-extrusion step, and a calcination in order to obtain the final alumina extrudates. Alumina is described as a complex porous medium, generally agglomerates constituted of aggregated nanocrystallites. Its porous texture directly influences the active phase deposition and mass transfer, and the catalytic properties. Then, it is easy to figure out that each step of the fabrication of the supports has a role on the building of their porous network, and has to be well understood to optimize the process. The synthesis of boehmite by precipitation of aluminum salts was extensively studied in the literature and the effect of various parameters, such as temperature or pH, are known to influence the size and shape of the crystallites and the specific surface area of the support. The calcination step, through the topotactic transition from boehmite to alumina, determines the final properties of the support and can tune the surface area, pore volume and pore diameters from those of boehmite. However, the kneading extrusion step has been subject to a very few studies. It generally consists in two steps: an acid, then a basic kneading, where the boehmite powder is introduced in a mixer and successively added with an acid and a base solution to form an extrudable paste. During the acid kneading, the induced positive charges on the hydroxyl surface groups of boehmite create an electrostatic repulsion which tends to separate the aggregates and even, following the conditions, the crystallites. The basic kneading, by reducing the surface charges, leads to a flocculation phenomenon and can control the reforming of the overall structure. The separation and reassembling of the particles constituting the boehmite paste have a quite obvious influence on the textural properties of the material. In this work, we are focused on the influence of the kneading step on the alumina catalysts supports. Starting from an industrial boehmite, extrudates are prepared using various kneading conditions. The samples are studied by nitrogen physisorption in order to analyze the evolution of the textural properties, and by synchrotron small-angle X-ray scattering (SAXS), a more original method which brings information about agglomeration and aggregation of the samples. The coupling of physisorption and SAXS enables a precise description of the samples, as same as an accurate monitoring of their evolution as a function of the kneading conditions. These ones are found to have a strong influence of the pore volume and pore size distribution of the supports. A mechanism of evolution of the texture during the kneading step is proposed and could be attractive in order to optimize the texture of the supports and then, their catalytic performances.

Keywords: alumina catalyst support, kneading, nitrogen physisorption, small-angle X-ray scattering

Procedia PDF Downloads 239

Authors: Modreck Gomo

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The evolution of groundwater chemistry and its quality is largely controlled by hydrogeochemical processes and their understanding is therefore important for groundwater quality assessments and protection of the water resources. A study was conducted in Bloemfontein town of South Africa to assess and compare the groundwater chemistry and quality characteristics in an alluvial aquifer and single-plane fractured-rock aquifers. 9 groundwater samples were collected from monitoring boreholes drilled into the two aquifer systems during a once-off sampling exercise. Samples were collected through low-flow purging technique and analysed for major ions and trace elements. In order to describe the hydrochemical facies and identify dominant hydrogeochemical processes, the groundwater chemistry data are interpreted using stiff diagrams and principal component analysis (PCA), as complimentary tools. The fitness of the groundwater quality for domestic and irrigation uses is also assessed. Results show that the alluvial aquifer is characterised by a Na-HCO₃ hydrochemical facie while fractured-rock aquifer has a Ca-HCO₃ facie. The groundwater in both aquifers originally evolved from the dissolution of calcite rocks that are common on land surface environments. However the groundwater in the alluvial aquifer further goes through another evolution as driven by cation exchange process in which Na in the sediments exchanges with Ca²⁺ in the Ca-HCO₃ hydrochemical type to result in the Na-HCO₃ hydrochemical type. Despite the difference in the hydrogeochemical processes between the alluvial aquifer and single-plane fractured-rock aquifer, this did not influence the groundwater quality. The groundwater in the two aquifers is very hard as influenced by the elevated magnesium and calcium ions that evolve from dissolution of carbonate minerals which typically occurs in surface environments. Based on total dissolved levels (600-900 mg/L), groundwater quality of the two aquifer systems is classified to be of fair quality. The negative potential impacts of the groundwater quality for domestic uses are highlighted.

Keywords: alluvial aquifer, fractured-rock aquifer, groundwater quality, hydrogeochemical processes

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Authors: Sairah Bashir, Liu Yang, John Liggat, James Thomason

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Glass fibre is currently used as reinforcement in over 90% of all fibre-reinforced composites produced. The high rigidity and chemical resistance of these composites are required for optimum performance but unfortunately results in poor recyclability; when such materials are no longer fit for purpose, they are frequently deposited in landfill sites. Recycling technologies, for example, thermal treatment, can be employed to address this issue; temperatures typically between 450 and 600 °C are required to allow degradation of the rigid polymeric matrix and subsequent extraction of fibrous reinforcement. However, due to the severe thermal conditions utilised in the recycling procedure, glass fibres become too weak for reprocessing in second-life composite materials. In addition, more stringent legislation is being put in place regarding disposal of composite waste, and so it is becoming increasingly important to develop long-term recycling solutions for such materials. In particular, the development of a cost-effective method to regenerate strength of thermally recycled glass fibres will have a positive environmental effect as a reduced volume of composite material will be destined for landfill. This research study has demonstrated the positive impact of sodium hydroxide (NaOH) and potassium hydroxide (KOH) solution, prepared at relatively mild temperatures and at concentrations of 1.5 M and above, on the strength of heat-treated glass fibres. As a result, alkaline treatments can potentially be implemented to glass fibres that are recycled from composite waste to allow their reuse in second-life materials. The optimisation of the strength recovery process is being conducted by varying certain reaction parameters such as molarity of alkaline solution and treatment time. It is believed that deep V-shaped surface flaws exist commonly on severely damaged fibre surfaces and are effectively removed to form smooth, U-shaped structures following alkaline treatment. Although these surface flaws are believed to be present on glass fibres they have not in fact been observed, however, they have recently been discovered in this research investigation through analytical techniques such as AFM (atomic force microscopy) and SEM (scanning electron microscopy). Reaction conditions such as molarity of alkaline solution affect the degree of etching of the glass fibre surface, and therefore the extent to which fibre strength is recovered. A novel method in determining the etching rate of glass fibres after alkaline treatment has been developed, and the data acquired can be correlated with strength. By varying reaction conditions such as alkaline solution temperature and molarity, the activation energy of the glass etching process and the reaction order can be calculated respectively. The promising results obtained from NaOH and KOH treatments have opened an exciting route to strength regeneration of thermally recycled glass fibres, and the optimisation of the alkaline treatment process is being continued in order to produce recycled fibres with properties that match original glass fibre products. The reuse of such glass filaments indicates that closed-loop recycling of glass fibre reinforced composite (GFRC) waste can be achieved. In fact, the development of a closed-loop recycling process for GFRC waste is already underway in this research study.

Keywords: glass fibers, glass strengthening, glass structure and properties, surface reactions and corrosion

Procedia PDF Downloads 239

Authors: Yakup Ulusu, Numan Eczacıoğlu, İsa Gökçe, Helen Waller, Jeremy H. Lakey

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Besides having the appropriate amino acid sequence to perform the function of proteins, it is important to have correct conformation after this sequence to process. To consist of this conformation depends on the amino acid sequence at the primary structure, hydrophobic interaction, chaperones and enzymes in charge of folding etc. Misfolded proteins are not functional and tend to be aggregated. Cysteine originating disulfide cross-links make stable this conformation of functional proteins. When two of the cysteine amino acids come side by side, disulfide bond is established that forms a cystine bridge. Due to this feature cysteine plays an important role on the formation of three-dimensional structure of many proteins. There are two cysteine amino acids (C44, C69) in the Tol-A-III protein. Unlike protein disulfide bonds from within his own, any non-specific cystine bridge causes a change in the three dimensional structure of the protein. Proteins can be expressed in various host cells as directly or fusion (chimeric). As a result of overproduction of the recombinant proteins, aggregation of insoluble proteins in the host cell can occur by forming a crystal structure called inclusion body. In general fusion proteins are produced for provide affinity tags to make proteins more soluble and production of some toxic proteins via fusion protein expression system like pTolT. Proteins can be modified by using a site-directed mutagenesis. By this way, creation of non-specific disulfide crosslinks can be prevented at fusion protein expression system via the present cysteine replaced by another amino acid such as serine, glycine or etc. To do this, we need; a DNA molecule that contains the gene that encodes for the target protein, required primers for mutation to be designed according to site directed mutagenesis reaction. This study was aimed to be replaced cysteine encoding codon TGT with serine encoding codon AGT. For this sense and reverse primers designed (given below) and used site-directed mutagenesis reaction. Several new copy of the template plasmid DNA has been formed with above mentioned mutagenic primers via polymerase chain reaction (PCR). PCR product consists of both the master template DNA (wild type) and the new DNA sequences containing mutations. Dpn-l endonuclease restriction enzyme which is specific for methylated DNA and cuts them to the elimination of the master template DNA. E. coli cells obtained after transformation were incubated LB medium with antibiotic. After purification of plasmid DNA from E. coli, the presence of the mutation was determined by DNA sequence analysis. Developed this new plasmid is called PtolT-δ.

Keywords: site directed mutagenesis, Escherichia coli, pTolT, protein expression

Procedia PDF Downloads 349