Search results for: liquid chromatography

Authors: Zhuanfang Fred Zhang, Tim C. Johnson, Yilin Fang, Chris E. Strickland

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A variety of geophysical techniques are available to image the opaque subsurface with little or no contact with the soil. It is common to conduct time-lapse surveys of different types for a given site for improved results of subsurface imaging. Regardless of the chosen survey methods, it is often a challenge to process the massive amount of survey data. The currently available software applications are generally based on the one-dimensional assumption for a desktop personal computer. Hence, they are usually incapable of imaging the three-dimensional (3D) processes/variables in the subsurface of reasonable spatial scales; the maximum amount of data that can be inverted simultaneously is often very small due to the capability limitation of personal computers. Presently, high-performance or integrating software that enables real-time integration of multi-process geophysical methods is needed. E4D-MP enables the integration and inversion of time-lapsed large-scale data surveys from geophysical methods. Using the supercomputing capability and parallel computation algorithm, E4D-MP is capable of processing data across vast spatiotemporal scales and in near real time. The main code and the modules of E4D-MP for inverting individual or combined data sets of time-lapse 3D electrical resistivity, spectral induced polarization, and gravity surveys have been developed and demonstrated for sub-surface imaging. E4D-MP provides capability of imaging the processes (e.g., liquid or gas flow, solute transport, cavity development) and subsurface properties (e.g., rock/soil density, conductivity) critical for successful control of environmental engineering related efforts such as environmental remediation, carbon sequestration, geothermal exploration, and mine land reclamation, among others.

Keywords: gravity survey, high-performance computing, sub-surface monitoring, electrical resistivity tomography

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Authors: Prabhat Kashyap, Krishan Kumar

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Delhi, the capital of India, is one of the most populated and polluted city among the world. In terms of air quality, Delhi’s air is degrading day by day & becomes worst of any major city in the world. The role of biogenic volatile organic compounds (BVOCs) is not much studied in cities like Delhi as a culprit for degraded air quality. They not only play a critical role in rural areas but also determine the atmospheric chemistry of urban areas as well. Particularly, Isoprene (2-methyl 1,3-butadiene, C5H8) is the single largest emitted compound among other BVOCs globally, that influence the tropospheric ozone chemistry in urban environment as the ozone forming potential of isoprene is very high. It is mainly emitted by vegetation & a small but significant portion is also released by vehicular exhaust of petrol operated vehicles. This study investigates the spatial and temporal variations of quantitative measurements of isoprene emissions along with different traffic tracers in 2 different seasons (post-monsoon & winter) at four different locations of Delhi. For the quantification of anthropogenic and biogenic isoprene, two sites from traffic intersections (Punjabi Bagh & CRRI) and two sites from vegetative locations (JNU & Yamuna Biodiversity Park) were selected in the vicinity of isoprene emitting tree species like Ficus religiosa, Dalbergia sissoo, Eucalyptus species etc. The concentrations of traffic tracers like benzene, toluene were also determined & their robust ratios with isoprene were used to differentiate anthropogenic isoprene with biogenic portion at each site. The ozone forming potential (OFP) of all selected species along with isoprene was also estimated. For collection of intra-day samples (3 times a day) in each season, a pre-conditioned fenceline monitoring (FLM) carbopack X thermal desorption tubes were used and further analysis was done with Gas chromatography attached with mass spectrometry (GC-MS). The results of the study proposed that the ambient air isoprene is always higher in post-monsoon season as compared to winter season at all the sites because of high temperature & intense sunlight. The maximum isoprene emission flux was always observed during afternoon hours in both seasons at all sites. The maximum isoprene concentration was found to be 13.95 ppbv at Biodiversity Park during afternoon time in post monsoon season while the lower concentration was observed as low as 0.07 ppbv at the same location during morning hours in winter season. OFP of isoprene at vegetation sites is very high during post-monsoon because of high concentrations. However, OFP for other traffic tracers were high during winter seasons & at traffic locations. Furthermore, high correlation between isoprene emissions with traffic volume at traffic sites revealed that a noteworthy share of its emission also originates from road traffic.

Keywords: biogenic VOCs, isoprene emission, anthropogenic isoprene, urban vegetation

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Authors: Lorenzo Cocola, Massimo Fedel, Dragiša Savić, Bojana Danilović, Luca Poletto

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An instrument for the detection and evaluation of gaseous carbon dioxide in the headspace of closed containers has been developed in the context of Packsensor Italian-Serbian joint project. The device is based on Tunable Diode Laser Absorption Spectroscopy (TDLAS) with a Wavelength Modulation Spectroscopy (WMS) technique in order to accomplish a non-invasive measurement inside closed containers of fermented dairy products (yogurts and fermented cheese in cups and bottles). The purpose of this instrument is the continuous monitoring of carbon dioxide concentration during incubation and storage of products over a time span of the whole shelf life of the product, in the presence of different microorganisms. The instrument’s optical front end has been designed to be integrated in a thermally stabilized incubator. An embedded computer provides processing of spectral artifacts and storage of an arbitrary set of calibration data allowing a properly calibrated measurement on many samples (cups and bottles) of different shapes and sizes commonly found in the retail distribution. A calibration protocol has been developed in order to be able to calibrate the instrument on the field also on containers which are notoriously difficult to seal properly. This calibration protocol is described and evaluated against reference measurements obtained through an industry standard (sampling) carbon dioxide metering technique. Some sets of validation test measurements on different containers are reported. Two test recordings of carbon dioxide concentration evolution are shown as an example of instrument operation. The first demonstrates the ability to monitor a rapid yeast growth in a contaminated sample through the increase of headspace carbon dioxide. Another experiment shows the dissolution transient with a non-saturated liquid medium in presence of a carbon dioxide rich headspace atmosphere.

Keywords: TDLAS, carbon dioxide, cups, headspace, measurement

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Authors: Ching Shya Lee, Mohamed Kheireddine Aroua, Wan Mohd Ashri Wan Daud, Patrick Cognet, Yolande Peres, Mohammed Ajeel

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In recent years, with the decreasing supply of fossil fuel, renewable energy has received a significant demand. Biodiesel which is well known as vegetable oil based fatty acid methyl ester is an alternative fuel for diesel. It can be produced from transesterification of vegetable oils, such as palm oil, sunflower oil, rapeseed oil, etc., with methanol. During the transesterification process, crude glycerol is formed as a by-product, resulting in 10% wt of the total biodiesel production. To date, due to the fast growing of biodiesel production in worldwide, the crude glycerol supply has also increased rapidly and resulted in a significant price drop for glycerol. Therefore, extensive research has been developed to use glycerol as feedstock to produce various added-value chemicals, such as tartronic acid, mesoxalic acid, glycolic acid, glyceric acid, propanediol, acrolein etc. The industrial processes that usually involved are selective oxidation, biofermentation, esterification, and hydrolysis. However, the conversion of glycerol into added-value compounds by electrochemical approach is rarely discussed. Currently, the approach is mainly focused on the electro-oxidation study of glycerol under potentiostatic mode for cogenerating energy with other chemicals. The electro-organic synthesis study from glycerol under galvanostatic mode is seldom reviewed. In this study, the glycerol was converted into various added-value compounds by electrochemical method under galvanostatic mode. This work aimed to study the possible compounds produced from glycerol by electrochemical technique in a one-pot electrolysis cell. The electro-organic synthesis study from glycerol was carried out in a single compartment reactor for 8 hours, over the platinum cathode and anode electrodes under acidic condition. Various parameters such as electric current (1.0 A to 3.0 A) and reaction temperature (27 °C to 80 °C) were evaluated. The products obtained were characterized by using gas chromatography-mass spectroscopy equipped with an aqueous-stable polyethylene glycol stationary phase column. Under the optimized reaction condition, the glycerol conversion achieved as high as 95%. The glycerol was successfully converted into various added-value chemicals such as ethylene glycol, glycolic acid, glyceric acid, acetaldehyde, formic acid, and glyceraldehyde; given the yield of 1%, 45%, 27%, 4%, 0.7% and 5%, respectively. Based on the products obtained from this study, the reaction mechanism of this process is proposed. In conclusion, this study has successfully converted glycerol into a wide variety of added-value compounds. These chemicals are found to have high market value; they can be used in the pharmaceutical, food and cosmetic industries. This study effectively opens a new approach for the electrochemical conversion of glycerol. For further enhancement on the product selectivity, electrode material is an important parameter to be considered.

Keywords: biodiesel, glycerol, electrochemical conversion, galvanostatic mode

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Authors: Tomasz Borowski, Dawid Sołoducha, Agata Markowska-Szczupak, Aneta Wesołowska, Marian Kordas, Rafał Rakoczy

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Essential oils (EOs) are fragrant volatile oils obtained from plants. These are used for cooking (for flavor and aroma), cleaning, beauty (e.g., rosemary essential oil is used to promote hair growth), health (e.g. thyme essential oil cures arthritis, normalizes blood pressure, reduces stress on the heart, cures chest infection and cough) and in the food industry as preservatives and antioxidants. Rosemary and thyme essential oils are considered the most eminent herbs based on their history and medicinal properties. They possess a wide range of activity against different types of bacteria and fungi compared with the other oils in both in vitro and in vivo studies. However, traditional uses of EOs are limited due to rosemary and thyme oils in high concentrations can be toxic. In light of the accessible data, the following hypothesis was put forward: Low frequency rotating magnetic field (RMF) increases the antimicrobial potential of EOs. The aim of this work was to investigate the antimicrobial activity of commercial Salvia Rosmarinus L. and Thymus vulgaris L. essential oil from Polish company Avicenna-Oil under Rotating Magnetic Field (RMF) at f = 25 Hz. The self-constructed reactor (MAP) was applied for this study. The chemical composition of oils was determined by gas chromatography coupled with mass spectrometry (GC-MS). Model bacteria Escherichia coli K12 (ATCC 25922) was used. Minimum inhibitory concentrations (MIC) against E. coli were determined for the essential oils. Tested oils in very small concentrations were prepared (from 1 to 3 drops of essential oils per 3 mL working suspensions). From the results of disc diffusion assay and MIC tests, it can be concluded that thyme oil had the highest antibacterial activity against E. coli. Moreover, the study indicates the exposition to the RMF, as compared to the unexposed controls causing an increase in the efficacy of antibacterial properties of tested oils. The extended radiation exposure to RMF at the frequency f= 25 Hz beyond 160 minutes resulted in a significant increase in antibacterial potential against E. coli. Bacteria were killed within 40 minutes in thyme oil in lower tested concentration (1 drop of essential oils per 3 mL working suspension). Rapid decrease (>3 log) of bacteria number was observed with rosemary oil within 100 minutes (in concentration 3 drops of essential oils per 3 mL working suspension). Thus, a method for improving the antimicrobial performance of essential oil in low concentrations was developed. However, it still remains to be investigated how bacteria get killed by the EOs treated by an electromagnetic field. The possible mechanisms relies on alteration in the permeability of ionic channels in ionic channels in the bacterial cell walls that transport in the cells was proposed. For further studies, it is proposed to examine other types of essential oils and other antibiotic-resistant bacteria (ARB), which are causing a serious concern throughout the world.

Keywords: rotating magnetic field, rosemary, thyme, essential oils, Escherichia coli

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Authors: Muhammad Abid

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The ultimate goal of this research was to produce a cold galvanizing compound (CGC) at reduced pigment volume concentration (PVC) to protect metallic structures from corrosion. The influence of the partial replacement of Zn dust by nano-layered graphene (NGr) and Zn metal nanoparticles on the electrochemical, morphological, rheological, and mechanical properties of CGC was investigated. EIS was used to explore the electrochemical nature of coatings. The EIS results revealed that the partial replacement of Zn by NGr and Zn nanoparticles enhanced the cathodic protection at reduced PVC (4:1) by improving the electrical contact between the Zn particles and the metal substrate. The Tafel scan was conducted to support the cathodic behaviour of the coatings. The sample formulated solely with Zn at PVC 4:1 was found to be dominated in physical barrier characteristics over cathodic protection. By increasing the concentration of NGr in the formulation, the corrosion potential shifted towards a more negative side. The coating with 1.5% NGr showed the highest galvanic action at reduced PVC. FE-SEM confirmed the interconnected network of conducting particles. The coating without NGr and Zn nanoparticles at PVC 4:1 showed significant gaps between the Zn dust particles. The novelty was evidenced when micrographs showed the consistent distribution of NGr and Zn nanoparticles all over the surface, which acted as a bridge between spherical Zn particles and provided cathodic protection at a reduced PVC. The layered structure of graphene also improved the physical shielding effect of the coatings, which limited the diffusion of electrolytes and corrosion products (oxides/hydroxides) into the coatings, which was reflected by the salt spray test. The rheological properties of coatings showed good liquid/fluid properties. All the coatings showed excellent adhesion but had different strength values. A real-time scratch resistance assessment showed all the coatings had good scratch resistance.

Keywords: protective coatings, anti-corrosion, galvanization, graphene, nanomaterials, polymers

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Authors: Matheus Müller, Delma Vidal

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The present paper aims to expose two techniques of dewatering for sludge, analyzing its operations and dewatering processes, aiming at improving the conditions of disposal of residues with high liquid content. It describes the field tests performed on two geotextile systems, a closed geotextile tube and an open geotextile drying bed, both of which are submitted to two filling cycles. The sludge used in the filling cycles for the field trials is from the water treatment plant of the Technological Center of Aeronautics – CTA, in São José dos Campos, Brazil. Data about volume and height abatement due to the dewatering and consolidation were collected per time, until it was observed constancy. With the laboratory analysis of the sludge allied to the data collected in the field, it was possible to perform a critical comparative study between the observed and the scientific literature, in this way, this paper expresses the data obtained and compares them with the bibliography. The tests were carried out on three fronts: field tests, including the filling cycles of the systems with the sludge from CTA, taking measurements of filling time per cycle and maximum filling height per cycle, heights against the abatement by dewatering of the systems over time; tests carried out in the laboratory, including the characterization of the sludge and removal of material samples from the systems to ascertain the solids content within the systems per time and; comparing the data obtained in the field and laboratory tests with the scientific literature. Through the study, it was possible to perceive that the process of densification of the material inside a closed system, such as the geotextile tube, occurs faster than the observed in the drying bed system. This process of accelerated densification can be brought about by the pumping pressure of the sludge in its filling and by the confinement of the residue through the permeable geotextile membrane (allowing water to pass through), accelerating the process of densification and dewatering by its own weight after the filling with sludge.

Keywords: consolidation, dewatering, geotextile drying bed, geotextile tube

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Authors: Olympia Nisiforou

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The cultural heritage of each country represents a unique and irreplaceable witness of the past. Nevertheless, on many occasions, such heritage is extremely vulnerable to natural disasters and reckless behaviors. Even if such exhibits are now located in Museums, they still receive insufficient protection due to improper environmental conditions. These external changes can negatively affect the conditions of the exhibits and contribute to inefficient maintenance in time. Hence, it is imperative to develop an innovative, low-cost system, to monitor indoor air quality systematically, since conventional methods are quite expensive and time-consuming. The present study gives an insight into the indoor air quality of the National Byzantine Museum of Cyprus. In particular, systematic measurements of particulate matter, bio-aerosols, the concentration of targeted chemical pollutants (including Volatile organic compounds (VOCs), temperature, relative humidity, and lighting conditions as well as microbial counts have been performed using conventional techniques. Measurements showed that most of the monitored physiochemical parameters did not vary significantly within the various sampling locations. Seasonal fluctuations of ammonia were observed, showing higher concentrations in the summer and lower in winter. It was found that the outdoor environment does not significantly affect indoor air quality in terms of VOC and Nitrogen oxides (NOX). A cutting-edge portable Gas Chromatography-Mass Spectrometry (GC-MS) system (TORION T-9) was used to identify and measure the concentrations of specific Volatile and Semi-volatile Organic Compounds. A large number of different VOCs and SVOCs found such as Benzene, Toluene, Xylene, Ethanol, Hexadecane, and Acetic acid, as well as some more complex compounds such as 3-ethyl-2,4-dimethyl-Isopropyl alcohol, 4,4'-biphenylene-bis-(3-aminobenzoate) and trifluoro-2,2-dimethylpropyl ester. Apart from the permanent indoor/outdoor sources (i.e., wooden frames, painted exhibits, carpets, ventilation system and outdoor air) of the above organic compounds, the concentration of some of them within the areas of the museum were found to increase when large groups of visitors were simultaneously present at a specific place within the museum. The high presence of Particulate Matter (PM), fungi and bacteria were found in the museum’s areas where carpets were present but low colonial counts were found in rooms where artworks are exhibited. Measurements mentioned above were used to validate an innovative low-cost air-quality monitoring system that has been developed within the present work. The developed system is able to monitor the average concentrations (on a bidaily basis) of several pollutants and presents several innovative features, including the prompt alerting in case of increased average concentrations of monitored pollutants, i.e., exceeding the limit values defined by the user.

Keywords: exibitions, indoor air quality , VOCs, pollution

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Authors: Malkhaz Jokhadze, Vakhtang Mshvildadze, Levan Makaradze, Ekaterine Mosidze, Salome Barbaqadze, Mariam Murtazashvili, Dali Berashvili, Koba sivsivadze, Lasha Bakuridze, Aliosha Bakuridze

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Essential oils are one of the most important groups of biologically active substances present in plants. Due to the chemical diversity of components, essential oils and their preparations have a wide spectrum of pharmacological action. They have bactericidal, antiviral, fungicidal, antiprotozoal, anti-inflammatory, spasmolytic, sedative and other activities. They are expectorant, spasmolytic, sedative, hypotensive, secretion enhancing, antioxidant remedies. Based on preliminary pharmacological studies, we have developed a formulation called “Phytobiotic” containing essential oils, a feed additive for poultry as an alternative to antibiotics. Phytobiotic is a water-soluble powder containing a composition of essential oils of thyme, clary, monarda and auxiliary substances: dry extract of liquorice and inhalation lactose. On this stage of research, the goal was to study the chemical composition of provided phytobiotic, identify the main substances and determine their quantity, investigate the biological activity of phytobiotic through in vitro and in vivo studies. Using gas chromatography-mass spectrometry, 38 components were identified in phytobiotic, representing acyclic-, monocyclic-, bicyclic-, and sesquiterpenes. Together with identification of main active substances, their quantitative content was determined, including acyclic terpene alcohol β-linalool, acyclic terpene ketone linalyl acetate, monocyclic terpenes: D-limonene and γ-terpinene, monocyclic aromatic terpene thymol. Provided phytobiotic has pronounced and at the same time broad spectrum of antibacterial activity. In the cell model, phytobiotic showed weak antioxidant activity, and it was stronger in the ORAC (chemical model) tests. Meanwhile anti-inflammatory activity was also observed. When fowls were supplied feed enriched with phytobiotic, it was observed that gained weight of the chickens in the experimental group exceeded the same data for the control group during the entire period of the experiment. The survival rate of broilers in the experimental group during the growth period was 98% compared to -94% in the control group. As a result of conducted researches probable four different mechanisms which are important for the action of phytobiotics were identified: sensory, metabolic, antioxidant and antibacterial action. General toxic, possible local irritant and allergenic effects of phytobiotic were also investigated. Performed assays proved that formulation is safe.

Keywords: clary, essential oils, monarda, poultry, phytobiotics, thyme

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Authors: Ganeshkumar, Ashika, Valavan, Ramesh, Thangam, Chirayu

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MSCs are found in much higher concentration in the Wharton’s jelly compared to the umbilical cord blood, which is a rich source of hematopoietic stem cells. Umbilical cord tissue is collected at the time of birth; it is processed and stored in liquid nitrogen for future therapeutical purpose. The source of contamination might be either from vaginal tract of mother or from hospital environment or from personal handling during cord tissue sample collection. If the sample were contaminated, decontamination procedure will be done with 70% ethanol (1 minute) in order to avoid sample rejection. Ethanol is effective against a wide range of bacteria, protozoa and fungi and has low toxicity to humans. Among the 1954 samples taken for the study, 24 samples were found to be contaminated with microorganism. The organisms isolated from the positive samples were found to be E. coli, Stenotrophomonas maltophilia, Pseudomonas aueroginosa, Enterococcus fecalis, Acinetobacter bowmani, Staphylococcus epidermidis, Enterobacter cloacae, and Proteus mirabilis. Among these organisms 70% ethanol successfully eliminated E. coli, Enterococcus fecalis, Acinetobacter bowmani, Staphylococcus epidermidis, and Proteus mirabilis. 70% ethanol was unsuccessful in eliminating Stenotrophomonas maltophilia, Pseudomonas aueroginosa, and Enterobacter cloacae. Stenotrophomonas maltophilia and Pseudomonas aueroginosa have the ability to form biofilm that make them resistant to alcohol. Biofilm act as protective layer for bacteria and which protects them from host defense and antibiotic wash. Finally it was found 70% ethanol wash saved 58.3% cord tissue samples from rejection and it is ineffective against 41% of the samples. The contamination rate can be reduced by maintaining proper aseptic techniques during sample collection and processing.

Keywords: umblical cord tissue, decontamination, 70% ethanol effectiveness, contamination

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Authors: Kira Toxopeus, Kamran Siddiqui

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Channel flows are common in a wide range of engineering applications. In some types of channel flows, particularly the ones involving chemical or biological processes, the control of the flow temperature is crucial to maintain the optimal conditions for the chemical reaction or to control the growth of biological species. This often becomes an issue when the flow experiences temperature fluctuations due to external conditions. While active heating and cooling could regulate the channel temperature, it may not be feasible logistically or economically and is also regarded as a non-sustainable option. Thermal energy storage utilizing phase change material (PCM) could provide the required thermal regulation sustainably by storing the excess heat from the channel and releasing it back as required, thus regulating the channel temperature within a range in the proximity of the PCM melting temperature. However, in designing such systems, the configuration of the PCM storage within the channel is critical as it could influence the channel flow dynamics, which would, in turn, affect the heat exchange between the channel fluid and the PCM. The present research is focused on the investigation of the flow dynamical behavior in the channel during heat transfer from the channel flow to the PCM thermal energy storage. Offset vertical columns in a narrow channel were used that contained the PCM. Two different column shapes, square and circular, were considered. Water was used as the channel fluid that entered the channel at a temperature higher than that of the PCM melting temperature. Hence, as the water was passing through the channel, the heat was being transferred from the water to the PCM, causing the PCM to store the heat through a phase transition from solid to liquid. Particle image velocimetry (PIV) was used to measure the two-dimensional velocity field of the channel flow as it flows between the PCM columns. Thermocouples were also attached to the PCM columns to measure the PCM temperature at three different heights. Three different water flow rates (0.5, 0.75 and 1.2 liters/min) were considered. At each flow rate, experiments were conducted at three different inlet water temperatures (28ᵒC, 33ᵒC and 38ᵒC). The results show that the flow rate and the inlet temperature influenced the flow behavior inside the channel.

Keywords: channel flow, phase change material, thermal energy storage, thermal regulation

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Authors: Mathew Saxon A, Aneeh Rajan, Sajeev P

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Many fluid flow applications employ different types of orifices to control the flow rate or to reduce the pressure. Discharge coefficients generally vary from 0.6 to 0.95 depending on the type of the orifice. The tabulated value of discharge coefficients of various types of orifices available can be used in most common applications. The upstream and downstream flow condition of an orifice is hardly considered while choosing the discharge coefficient of an orifice. But literature shows that the discharge coefficient can be affected by the presence of cross flow. Cross flow is defined as the condition wherein; a fluid is injected nearly perpendicular to a flowing fluid. Most researchers have worked on water being injected into a cross-flow of water. The present work deals with water to gas systems in which water is injected in a normal direction into a flowing stream of gas. The test article used in the current work is called thermal regulator, which is used in a liquid rocket engine to reduce the temperature of hot gas tapped from the gas generator by injecting water into the hot gas so that a cooler gas can be supplied to the turbine. In a thermal regulator, water is injected through an orifice in a normal direction into the hot gas stream. But the injection orifice had been calibrated under backpressure by maintaining a stagnant gas medium at the downstream. The motivation of the present study aroused due to the observation of a lower Cd of the orifice in flight compared to the calibrated Cd. A systematic experimental investigation is carried out in this paper to study the effect of cross-flow on the discharge coefficient of an orifice in water to a gas system. The study reveals that there is an appreciable reduction in the discharge coefficient with cross flow compared to that without cross flow. It is found that the discharge coefficient greatly depends on the ratio of momentum of water injected to the momentum of the gas cross flow. The effective discharge coefficient of different orifices was normalized using the discharge coefficient without cross-flow and it is observed that normalized curves of effective discharge coefficient of different orifices with momentum ratio collapsing into a single curve. Further, an equation is formulated using the test data to predict the effective discharge coefficient with cross flow using the calibrated Cd value without cross flow.

Keywords: cross flow, discharge coefficient, orifice, momentum ratio

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Authors: Cristian Soitu, Alexander Feuerborn, Cyril Deroy, Alfonso Castrejon-Pita, Peter R. Cook, Edmond J. Walsh

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Microfluidics now stands as an academically mature technology after a quarter of a century research activities have delivered a vast array of proof of concepts for many biological workflows. However, translation to industry remains poor, with only a handful of notable exceptions – e.g. digital PCR, DNA sequencing – mainly because of biocompatibility issues, limited range of readouts supported or complex operation required. This technology exploits the domination of interfacial forces over gravitational ones at the microscale, replacing solid walls with fluid ones as building blocks for cell micro-environments. By employing only materials used by biologists for decades, the system is shown to be biocompatible, and easy to manufacture and operate. The method consists in displacing a continuous fluid layer into a pattern of isolated chambers overlaid with an immiscible liquid to prevent evaporation. The resulting fluid arrangements can be arrays of micro-chambers with rectangular footprint, which use the maximum surface area available, or structures with irregular patterns. Pliant, self-healing fluid walls confine volumes as small as 1 nl. Such fluidic structures can be reconfigured during the assays, giving the platform an unprecedented level of flexibility. Common workflows in cell biology are demonstrated – e.g. cell growth and retrieval, cloning, cryopreservation, fixation and immunolabeling, CRISPR-Cas9 gene editing, and proof-of-concept drug tests. This fluid-shaping technology is shown to have potential for high-throughput cell- and organism-based assays. The ability to make and reconfigure on-demand microfluidic circuits on standard Petri dishes should find many applications in biology, and yield more relevant phenotypic and genotypic responses when compared to standard microfluidic assays.

Keywords: fluid walls, micro-chambers, reconfigurable, freestyle

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Authors: Abhishek N. Srivastava, Rahul Singh, Sumedha Chakma

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The rapid urbanization in the developing countries is generating an enormous amount of waste leading to the creation of unregulated landfill sites at various places at its disposal. The liquid waste, known as leachate, produced from these landfills sites is severely affecting the surrounding water quality. The water quality in the proximity areas of the landfill is found affected by various physico-chemical parameters of leachate such as pH, alkalinity, total hardness, conductivity, chloride, total dissolved solids (TDS), total suspended solids (TSS), sulphate, nitrate, phosphate, fluoride, sodium and potassium, biological parameters such as biochemical oxygen demand (BOD), chemical oxygen demand (COD), Faecal coliform, and heavy metals such as cadmium (Cd), lead (Pb), iron (Fe), mercury (Hg), arsenic (As), cobalt (Co), manganese (Mn), zinc (Zn), copper (Cu), chromium (Cr), nickel (Ni). However, all these parameters are distributive in leachate that produced according to the nature of waste being dumped at various landfill sites, therefore, it becomes very difficult to predict the main responsible parameter of leachate for water quality contamination. The present study is endeavour the comparative analysis of the physical, chemical and biological parameters of various landfills in India viz. Okhla landfill, Ghazipur landfill, Bhalswa ladfill in NCR Delhi, Deonar landfill in Mumbai, Dhapa landfill in Kolkata and Kodungayaiyur landfill, Perungudi landfill in Chennai. The statistical analysis of the parameters was carried out using the Statistical Packages for the Social Sciences (SPSS) and LandSim 2.5 model to simulate the long term effect of various parameters on different time scale. Further, the uncertainties characterization of various input parameters has also been analysed using fuzzy alpha cut (FAC) technique to check the sensitivity of various water quality parameters at the proximity of numerous landfill sites. Finally, the study would help to suggest the best method for the prevention of pollution migration from the landfill sites on priority basis.

Keywords: landfill leachate, water quality, LandSim, fuzzy alpha cut

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Authors: Paulomi Polly Burey, Mark Lynch

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It has been shown that laboratory activities can help cement understanding of theoretical concepts, but it is difficult to deliver such an activity to an online cohort and issues such as occupational health and safety in the students’ learning environment need to be considered. Chemistry, in particular, is one of the sciences where practical experience is beneficial for learning, however typical university experiments may not be suitable for the learning environment of a distance learner. Food provides an ideal medium for demonstrating chemical concepts, and along with a few simple physical and virtual tools provided by educators, analytical chemistry can be experienced by distance learners. Food chemistry experiments were designed to be carried out in a home-based environment that 1) Had sufficient scientific rigour and skill-building to reinforce theoretical concepts; 2) Were safe for use at home by university students and 3) Had the potential to enhance student learning by linking simple hands-on laboratory activities with high-level virtual science. Two main components of the resources were developed, a home laboratory experiment component, and a virtual laboratory component. For the home laboratory component, students were provided with laboratory kits, as well as a list of supplementary inexpensive chemical items that they could purchase from hardware stores and supermarkets. The experiments used were typical proximate analyses of food, as well as experiments focused on techniques such as spectrophotometry and chromatography. Written instructions for each experiment coupled with video laboratory demonstrations were used to train students on appropriate laboratory technique. Data that students collected in their home laboratory environment was collated across the class through shared documents, so that the group could carry out statistical analysis and experience a full laboratory experience from their own home. For the virtual laboratory component, students were able to view a laboratory safety induction and advised on good characteristics of a home laboratory space prior to carrying out their experiments. Following on from this activity, students observed laboratory demonstrations of the experimental series they would carry out in their learning environment. Finally, students were embedded in a virtual laboratory environment to experience complex chemical analyses with equipment that would be too costly and sensitive to be housed in their learning environment. To investigate the impact of the intervention, students were surveyed before and after the laboratory series to evaluate engagement and satisfaction with the course. Students were also assessed on their understanding of theoretical chemical concepts before and after the laboratory series to determine the impact on their learning. At the end of the intervention, focus groups were run to determine which aspects helped and hindered learning. It was found that the physical experiments helped students to understand laboratory technique, as well as methodology interpretation, particularly if they had not been in such a laboratory environment before. The virtual learning environment aided learning as it could be utilized for longer than a typical physical laboratory class, thus allowing further time on understanding techniques.

Keywords: chemistry, food science, future pedagogy, STEM education

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Authors: E.K.K. Agyeman, P. Mousseau, A. Sarda, D. Edelin

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The uniform and controlled cooling of hot metals by the circulation of water in canals remains a challenge due to the phase change of the water and the high heat fluxes associated with the phase change. This is because, during the cooling process, the phases are not uniformly distributed along the canals with the liquid phase dominating at the entrances of the canals and the gaseous phase dominating towards the exits. The difference in thermal properties between both phases leads to a heterogeneous temperature distribution in the part being cooled. Slowing down the cooling process is also a challenge due to the high heat fluxes associated with the phase change of water. This study investigates the use of multiple water jets for the controlled and homogenous cooling of hot metal parts and the effect of gravity on the effectiveness of the cooling process with a potential application in the cooling of composite forming moulds. A hole is bored at the centre of a steel block along its length. The jets are generated from the holes of a perforated steel pipe which is placed along the centre of the hole bored in the steel block. The evolution of the temperature with respect to time on the external surface of the steel block is measured simultaneously by thermocouples and an infrared camera. Different jet positions are tested in order to identify the jet placement configuration that ensures the most homogenous cooling of the block while the cooling speed is controlled by an intermittent impingement of the jets. In order to study the effect of gravity on the cooling process, a scenario where the jets are oriented in the opposite direction to that of gravity is compared to one where the jets are aligned in the same direction as gravity. It’s observed that orienting the jets in the direction of gravity reduces the effectiveness of the cooling process on the face of the block facing the impinging jets. This is due to the formation of a deeper pool of water due to the effect gravity and of the curved surface of the canal. This deeper pool of water influences the boiling regime characterized by a slower bubble evacuation when compared to the scenario where the jets are opposed to gravity.

Keywords: cooling speed, gravity, homogenous cooling, jet impingement

Procedia PDF Downloads 103

Authors: Suveen Kumar

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Oral cancer (OC) is the sixth most death causing cancer in world which includes tumour of lips, floor of the mouth, tongue, palate, cheeks, sinuses, throat, etc. Conventionally, the techniques used for OC detection are toluidine blue staining, biopsy, liquid-based cytology, visual attachments, etc., however these are limited by their highly invasive nature, low sensitivity, time consumption, sophisticated instrument handling, sample processing and high cost. Therefore, we developed biosensing chips for non-invasive detection of OC via CYFRA-21-1 biomarker. CYFRA-21-1 (molecular weight: 40 kDa) is secreted in saliva of OC patients which is a non-invasive biological fluid with a cut-off value of 3.8 ng mL-1, above which the subjects will be suffering from oral cancer. Therefore, in first work, 3-aminopropyl triethoxy silane (APTES) functionalized zirconia (ZrO2) nanoparticles (APTES/nZrO2) were used to successfully detect CYFRA-21-1 in a linear detection range (LDR) of 2-16 ng mL-1 with sensitivity of 2.2 µA mL ng-1. Successively, APTES/nZrO2-RGO was employed to prevent agglomeration of ZrO2 by providing high surface area reduced graphene oxide (RGO) support and much wider LDR (2-22 ng mL-1) was obtained with remarkable limit of detection (LOD) as 0.12 ng mL-1. Further, APTES/nY2O3/ITO platform was used for oral cancer bioseneor development. The developed biosensor (BSA/anti-CYFRA-21-1/APTES/nY2O3/ITO) have wider LDR (0.01-50 ng mL-1) with remarkable limit of detection (LOD) as 0.01 ng mL-1. To improve the sensitivity of the biosensing platform, nanocomposite of yattria stabilized nanostructured zirconia-reduced graphene oxide (nYZR) based biosensor has been developed. The developed biosensing chip having ability to detect CYFRA-21-1 biomolecules in the range of 0.01-50 ng mL-1, LOD of 7.2 pg mL-1 with sensitivity of 200 µA mL ng-1. Further, the applicability of the fabricated biosensing chips were also checked through real sample (saliva) analysis of OC patients and the obtained results showed good correlation with the standard protein detection enzyme linked immunosorbent assay (ELISA) technique.

Keywords: non-invasive, oral cancer, nanomaterials, biosensor, biochip

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Authors: Joana Antunes, Thomas Levée, Barbara Radovani, Anu Suonpää, Paulina Saloranta, Liji Sobhana, Petri Ihalainen

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Lignin is an important component in the exploitation of lignocellulosic biomass. It has been shown that within the next years, the yield of added-value lignin-based chemicals and materials will generate renewable alternatives to oil-based products (e.g. polymeric composites, resins and adhesives) and enhance the economic feasibility of biorefineries. In this paper, a novel technology for lignin valorisation (METNIN™) is presented. METNIN™ is based on the oxidative action of an alkaliphilic enzyme in aqueous alkaline conditions (pH 10-11) at mild temperature (40-50 °C) combined with a cascading membrane operation, yielding a collection of lignin fractions (from oligomeric down to mixture of tri-, di- and monomeric units) with distinct molecular weight distribution, low polydispersity and favourable physicochemical properties. The alkaline process conditions ensure the high processibility of crude lignin in an aqueous environment and the efficiency of the enzyme, yielding better compatibility of lignin towards targeted applications. The application of a selected lignin fraction produced by METNIN™ as a suitable lignopolyol to completely replace a commercial polyol in polyurethane rigid foam formulations is presented as a prototype. Liquid lignopolyols with a high lignin content were prepared by oxypropylation and their full utilization in the polyurethane rigid foam formulation was successfully demonstrated. Moreover, selected technical specifications of different foam demonstrators were determined, including closed cell count, water uptake and compression characteristics. These specifications are within industrial standards for rigid foam applications. The lignin loading in the lignopolyol was a major factor determining the properties of the foam. In addition to polyurethane foam demonstrators, other examples of lignin-based products related to resins and sizing applications will be presented.

Keywords: enzyme, lignin valorisation, polyol, polyurethane foam

Procedia PDF Downloads 123

Authors: Semenov Matvei, Stoporev Andrey, Pavelyev Roman, Varfolomeev Mikhail

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Gas hydrates are host-guest compounds. Guest molecules can be low molecular weight components of associated petroleum gas (C1-C4 hydrocarbons), carbon dioxide, hydrogen sulfide, and nitrogen. Gas hydrates have a number of unique properties that make them interesting from a technological point of view, for example, for storing hydrocarbon gases in solid form under moderate thermobaric conditions. The hydrate form of gas has a number of advantages, including a significant gas content in the hydrate, relative safety and environmental friendliness of the process. Such technology could be especially useful in cold regions, where hydrate production, storage and transportation can be more energy efficient. Recently, new developments have been proposed that seek to reduce the number of steps to obtain the finished hydrate, for example, using a pressing device/screw inside the reactor. However, the energy consumption required for the hydrate formation process remains a challenge. Thus, the goal of the current work is to study the patterns and mechanisms of the hydrate formation process using small additions of hydrate formation promoters under static conditions. The study of these aspects will help solve the problem of accelerated production of gas hydrates with minimal energy consumption. Currently, new compounds have been developed that can accelerate the formation of methane hydrate with a small amount of promoter in water, not exceeding 0.1% by weight. To test the influence of promoters on the process of hydrate formation, standard experiments are carried out under dynamic conditions with stirring. During such experiments, the time at which hydrate formation begins (induction period), the temperature at which formation begins (supercooling), the rate of hydrate formation, and the degree of conversion of water to hydrate are assessed. This approach helps to determine the most effective compound in comparative experiments with different promoters and select their optimal concentration. These experimental studies made it possible to study the features of the formation of associated petroleum gas hydrate from promoter solutions under static conditions. Phase transformations were studied using high-pressure micro-differential scanning calorimetry under various experimental conditions. Visual studies of the growth mode of methane hydrate depending on the type of promoter were also carried out. The work is an extension of the methodology for studying the effect of promoters on the process of associated petroleum gas hydrate formation in order to identify new ways to accelerate the formation of gas hydrates without the use of mixing. This work presents the results of a study of the process of associated petroleum gas hydrate formation using high-pressure differential scanning micro-calorimetry, visual investigation, gas chromatography, autoclaves study and stability data. It was found that the synthesized compounds multiply the conversion of water into hydrate under static conditions up to 96% due to a change in the growth mechanism of associated petroleum gas hydrate.

Keywords: gas hydrate, gas storage, promotor, associated petroleum gas

Procedia PDF Downloads 31

Authors: Hui Long, Chun Yin Tang, Ping Kwong Cheng, Xin Yu Wang, Wayesh Qarony, Yuen Hong Tsang

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Two dimensional (2D) materials have recently attained substantial research interest since the discovery of graphene. However, the zero-bandgap feature of the graphene limits its nonlinear optical applications, e.g., saturable absorption for these applications require strong light-matter interaction. Nevertheless, the excellent optoelectronic properties, such as broad tunable bandgap energy and high carrier mobility of Group 10 transition metal dichalcogenides 2D materials, e.g., PtS2 introduce new degree of freedoms in the optoelectronic applications. This work reports our recent research findings regarding the saturable absorption property of PtS2 layered 2D material and its possibility to be used as saturable absorber (SA) for ultrafast mode locking fiber laser. The demonstration of mode locking operation by using the fabricated PtS2 as SA will be discussed. The PtS2/PVA SA used in this experiment is made up of some few layered PtS2 nanosheets fabricated via a simple ultrasonic liquid exfoliation. The operational wavelength located at ~1 micron is demonstrated from Yb-doped mode locking fiber laser ring cavity by using the PtS2 SA. The fabricated PtS2 saturable absorber offers strong nonlinear properties, and it is capable of producing regular mode locking laser pulses with pulse to pulse duration matched with the round-trip cavity time. The results confirm successful mode locking operation achieved by the fabricated PtS2 material. This work opens some new opportunities for these PtS2 materials for the ultrafast laser generation. Acknowledgments: This work is financially supported by Shenzhen Science and Technology Innovation Commission (JCYJ20170303160136888) and the Research Grants Council of Hong Kong, China (GRF 152109/16E, PolyU code: B-Q52T).

Keywords: platinum disulfide, PtS2, saturable absorption, saturable absorber, mode locking laser

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Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

Procedia PDF Downloads 241

Authors: L. Asaro, M. Gratton, S. Seghar, N. Poirot, N. Ait Hocine

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Rubber waste disposal is an environmental problem. Particularly, many researches are centered in the management of discarded tires. In spite of all different ways of handling used tires, the most common is to deposit them in a landfill, creating a stock of tires. These stocks can cause fire danger and provide ambient for rodents, mosquitoes and other pests, causing health hazards and environmental problems. Because of the three-dimensional structure of the rubbers and their specific composition that include several additives, their recycling is a current technological challenge. The technique which can break down the crosslink bonds in the rubber is called devulcanization. Strictly, devulcanization can be defined as a process where poly-, di-, and mono-sulfidic bonds, formed during vulcanization, are totally or partially broken. In the recent years, super critical carbon dioxide (scCO₂) was proposed as a green devulcanization atmosphere. This is because it is chemically inactive, nontoxic, nonflammable and inexpensive. Its critical point can be easily reached (31.1 °C and 7.38 MPa), and residual scCO₂ in the devulcanized rubber can be easily and rapidly removed by releasing pressure. In this study thermomechanical devulcanization of ground tire rubber (GTR) was performed in a twin screw extruder under diverse operation conditions. Supercritical CO₂ was added in different quantities to promote the devulcanization. Temperature, screw speed and quantity of CO₂ were the parameters that were varied during the process. The devulcanized rubber was characterized by its devulcanization percent and crosslink density by swelling in toluene. Infrared spectroscopy (FTIR) and Gel permeation chromatography (GPC) were also done, and the results were related with the Mooney viscosity. The results showed that the crosslink density decreases as the extruder temperature and speed increases, and, as expected, the soluble fraction increase with both parameters. The Mooney viscosity of the devulcanized rubber decreases as the extruder temperature increases. The reached values were in good correlation (R= 0.96) with de the soluble fraction. In order to analyze if the devulcanization was caused by main chains or crosslink scission, the Horikx's theory was used. Results showed that all tests fall in the curve that corresponds to the sulfur bond scission, which indicates that the devulcanization has successfully happened without degradation of the rubber. In the spectra obtained by FTIR, it was observed that none of the characteristic peaks of the GTR were modified by the different devulcanization conditions. This was expected, because due to the low sulfur content (~1.4 phr) and the multiphasic composition of the GTR, it is very difficult to evaluate the devulcanization by this technique. The lowest crosslink density was reached with 1 cm³/min of CO₂, and the power consumed in that process was also near to the minimum. These results encourage us to do further analyses to better understand the effect of the different conditions on the devulcanization process. The analysis is currently extended to monophasic rubbers as ethylene propylene diene monomer rubber (EPDM) and natural rubber (NR).

Keywords: devulcanization, recycling, rubber, waste

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Authors: Aida Safiera, Andika Dwi Rubyantoro, Muhammad Bagus Prakasa

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Indonesia’s needs of diesel oil each year are increasing and getting urge. However, that problems are not supported by the amount of oil production that still low and also influenced by the fact of oil reserve is reduced. Because of that, the government prefers to import from other countries than fulfill the needs of diesel. To anticipate that problem, development of fuel based on renewable diesel is started. Renewable diesel is renewable alternative fuel that is hydrocarbon derivative from decarbonylation of non-edible oil. Indonesia is rich with natural resources, including nyamplung oil (Calophyllum inophyllum oil) and zeolite. Nyamplung oil (Calophyllum inophyllum oil) has many stearic acids which are useful on renewable diesel synthesis meanwhile zeolite is cheap. Zeolite is many used on high temperature reaction and cracking process on oil industry. Zeolite also has advantages which are a high crystallization, surface area and pores. In this research, the main focus that becomes our attention is on preparation and characterization of metal nanocrystal. Active site that used in this research is Nickel Molybdenum (NiMo). The advantage of nanocrystal with nano scale is having larger surface area. The synthesis of metal nanocrystal will be done with conventional preparation modification method that is called simple heating. Simple heating method is a metal nanocrystal synthesis method using continuous media which is polymer liquid. This method is a simple method and produces a small particles size in a short time. Influence of metal nanocrystal growth on this method is the heating profile. On the synthesis of nanocrystal, the manipulated variables are temperature and calcination time. Results to achieve from this research are diameter size on nano scale (< 100 nm) and uniform size without any agglomeration. Besides that, the conversion of synthesis of renewable diesel is high and has an equal specification with petroleum diesel. Catalyst activities are tested by FT-IR and GC-TCD on decarbonylation process with a pressure 15 bar and temperature 375 °C. The highest conversion from this reaction is 35% with selectivity around 43%.

Keywords: renewable diesel, simple heating, metal nanocrystal, NiMo, zeolite

Procedia PDF Downloads 203

Authors: Kathryn Rooney, Kaitlyn Eddy, Evan Landers, Weihui Li

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The international mortality rate for neonates and infants has been declining at a disproportionally low rate when compared to the overall decline in child mortality in recent decades. A significant portion of infant deaths could be prevented with the implementation of low-cost and easy to use physiological monitoring devices, by enabling early identification of symptoms before they progress into life-threatening illnesses. The oral diagnostic device discussed in this paper serves to continuously monitor the key vital signs of body temperature, respiratory rate, heart rate, and oxygen saturation. The device mimics an infant pacifier, designed to be easily tolerated by infants as well as orthodontically inert. The fundamental measurements are gathered via thermistors and a pulse oximeter, each encapsulated in medical-grade silicone and wired internally to a microcontroller chip. The chip then translates the raw measurements into physiological values via an internal algorithm, before outputting the data to a liquid crystal display screen and an Android application. Additionally, a biological sample collection chamber is incorporated into the internal portion of the device. The movement within the oral chamber created by sucking on the pacifier-like device pushes saliva through a small check valve in the distal end, where it is accumulated and stored. The collection chamber can be easily removed, making the sample readily available to be tested for various diseases and analytes. With the vital sign monitoring and sample collection offered by this device, abnormal fluctuations in physiological parameters can be identified and appropriate medical care can be sought. This device enables preventative diagnosis for infants who may otherwise have gone undiagnosed, due to the inaccessibility of healthcare that plagues vast numbers of underprivileged populations.

Keywords: neonate mortality, infant mortality, low-cost diagnostics, vital signs, saliva testing, preventative care

Procedia PDF Downloads 119

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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Authors: R. Fardid, R. Coppes

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Introduction: In mediastinal radiotherapy and to a lesser extend also in total-body irradiation (TBI) radiation exposure may lead to development of cardiac diseases. Radiation-induced heart disease is dose-dependent and it is characterized by a loss of cardiac function, associated with progressive heart cells degeneration. We aimed to determine the in-vivo radiation effects on fibronectin, ColaA1, ColaA2, galectin and TGFb1 gene expression levels in left ventricle heart tissues of rats after irradiation. Material and method: Four non-treatment adult Wistar rats as control group (group A) were selected. In group B, 4 adult Wistar rats irradiated to 20 Gy single dose of 150 Mev proton beam locally in heart only. In heart plus lung irradiate group (group C) 4 adult rats was irradiated by 50% of lung laterally plus heart radiation that mentioned in before group. At 8 weeks after radiation animals sacrificed and left ventricle heart dropped in liquid nitrogen for RNA extraction by Absolutely RNA® Miniprep Kit (Stratagen, Cat no. 400800). cDNA was synthesized using M-MLV reverse transcriptase (Life Technologies, Cat no. 28025-013). We used Bio-Rad machine (Bio Rad iQ5 Real Time PCR) for QPCR testing by relative standard curve method. Results: We found that gene expression of fibronectin in group C significantly increased compared to control group, but it was not showed significant change in group B compared to group A. The levels of gene expressions of Cola1 and Cola2 in mRNA did not show any significant changes between normal and radiation groups. Changes of expression of galectin target significantly increased only in group C compared to group A. TGFb1 expressions in group C more than group B showed significant enhancement compared to group A. Conclusion: In summary we can say that 20 Gy of proton exposure of heart tissue may lead to detectable damages in heart cells and may distribute function of them as a component of heart tissue structure in molecular level.

Keywords: gene expression, heart damage, proton irradiation, radiotherapy

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Authors: Neha Devi, P. K. Joshi

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Fire fuel map is one of the most critical factors for planning and managing the fire hazard and risk. One of the most significant forms of global disturbance, impacting community dynamics, biogeochemical cycles and local and regional climate across a wide range of ecosystems ranging from boreal forests to tropical rainforest is wildfire Assessment of fire danger is a function of forest type, fuelwood stock volume, moisture content, degree of senescence and fire management strategy adopted in the ground. Remote sensing has potential of reduction the uncertainty in mapping fuels. Hyperspectral remote sensing is emerging to be a very promising technology for wildfire fuels characterization. Fine spectral information also facilitates mapping of biophysical and chemical information that is directly related to the quality of forest fire fuels including above ground live biomass, canopy moisture, etc. We used Hyperion imagery acquired in February, 2016 and analysed four fuel characteristics using Hyperion sensor data on-board EO-1 satellite, acquired over the Shiwalik Himalayas covering the area of Champawat, Uttarakhand state. The main objective of this study was to present an overview of methodologies for mapping fuel properties using hyperspectral remote sensing data. Fuel characteristics analysed include fuel biomass, fuel moisture, and fuel condition and fuel type. Fuel moisture and fuel biomass were assessed through the expression of the liquid water bands. Fuel condition and type was assessed using green vegetation, non-photosynthetic vegetation and soil as Endmember for spectral mixture analysis. Linear Spectral Unmixing, a partial spectral unmixing algorithm, was used to identify the spectral abundance of green vegetation, non-photosynthetic vegetation and soil.

Keywords: forest fire fuel, Hyperion, hyperspectral, linear spectral unmixing, spectral mixture analysis

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Authors: Kirti Hira, A. Sajeli Begum, S. Mahibalan, Poorna Chandra Rao

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Introduction: Cancer is one of the leading causes of death worldwide. Although existing therapy effectively kills cancer cells, they do affect normal growing cells leading to many undesirable side effects. Hence there is need to develop effective as well as safe drug molecules to combat cancer, which is possible through phyto-research. The currently available plant-derived blockbuster drugs are the example for this. In view of this, an investigation was done to identify cytotoxic lead molecules from Hedyotis umbellata (Family Rubiaceae), a widely distributed weed in India. Materials and Methods: The methanolic extract of the whole plant of H. umbellata (MHU), prepared through Soxhlet extraction method was further fractionated with diethyl ether and n-butanol, successively. MHU, ether fraction (EMHU) and butanol fraction (BMHU) were lyophilized and were tested for the cytotoxic effect using 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay against non-small cell lung cancer (NSCLC) A549 cell lines. The potentially active EMHU was subjected to chromatographic purification using normal-phase silica columns, in order to isolate the responsible bioactive compounds. The isolated pure compounds were tested for their cytotoxic effect by MTT assay against A549 cells. Compound-3, which was found to be most active, was characterized using IR, 1H- and 13C-NMR and MS analysis. The study was further extended to decipher the mechanism of action of cytotoxicity of compound-3 against A549 cells through various in vitro cellular models. Cell cycle analysis was done using flow cytometry following PI (Propidium Iodide) staining. Protein analysis was done using Western blot technique. Results: Among MHU, EMHU, and BMHU, the non-polar fraction EMHU demonstrated a significant dose-dependent cytotoxic effect with IC50 of 67.7μg/ml. Chromatography of EMHU yielded seven compounds. MTT assay of isolated compounds explored compound-3 as potentially active one, which inhibited the growth of A549 cells with IC50value of 14.2μM. Further, compound-3 was identified as cedrelopsin, a coumarin derivative having molecular weight of 260. Results of in vitro mechanistic studies explained that cedrelopsin induced cell cycle arrest at G2/M phase and down-regulated the expression of G2/M regulatory proteins such as cyclin B1, cdc2, and cdc25C, dose dependently. This is the first report that explores the cytotoxic mechanism of cedrelopsin. Conclusion: Thus a potential small lead molecule, cedrelopsin isolated from H. umbellata, showing antiproliferative effect mediated by G2/M arrest in A549 cells was discovered. The effect of cedrelopsin against other cancer cell lines followed by in vivo studies can be performed in future to develop a new drug candidate.

Keywords: A549, cedrelopsin, G2/M phase, Hedyotis umbellata

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Authors: Boor Singh Lalia, Yarjan Abdul Samad, Raed Hashaikeh

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Three different kinds of solid polymer electrolytes were prepared using polyethylene oxide (PEO) as a base polymer, networked cellulose (NC) as a physical support and LiClO4 as a conductive salt for the electrolytes. Networked cellulose, a modified form of cellulose, is a biodegradable and environmentally friendly additive which provides a strong fibrous networked support for structural stability of the electrolytes. Although the PEO/NC/LiClO4 electrolyte retains its structural integrity and mechanical properties at 100oC as compared to pristine PEO-based polymer electrolytes, it suffers from poor ionic conductivity. To improve the room temperature conductivity of the electrolyte, PEO is replaced by the polyethylene glycol (PEG) which is a liquid phase that provides high mobility for Li+ ions transport in the electrolyte. PEG/NC/LiClO4 shows improvement in ionic conductivity compared to PEO/NC/LiClO4 at room temperature, but it is brittle and tends to form cracks during processing. An advanced solid polymer electrolyte with optimum ionic conductivity and mechanical properties is developed by using a ternary system: TEGDME/PEO/NC+LiClO4. At room temperature, this electrolyte exhibits an ionic conductivity to the order of 10-5 S/cm, which is very high compared to that of the PEO/LiClO4 electrolyte. Pristine PEO electrolytes start melting at 65 °C and completely lose its mechanical strength. Dynamic mechanical analysis of TEGDME: PEO: NC (70:20:10 wt%) showed an improvement of storage modulus as compared to the pristine PEO in the 60–120 °C temperature range. Also, with an addition of NC, the electrolyte retains its mechanical integrity at 100 oC which is beneficial for Li-ion battery operation at high temperatures. Differential scanning calorimetry (DSC) and thermal gravimetry analysis (TGA) studies revealed that the ternary polymer electrolyte is thermally stable in the lithium ion battery operational temperature range. As-prepared polymer electrolyte was used to assemble LiFePO4/ TEGDME/PEO/NC+LiClO4/Li half cells and their electrochemical performance was studied via cyclic voltammetry and charge-discharge cycling.

Keywords: solid polymer electrolyte, ionic conductivity, mechanical properties, lithium ion batteries, cyclic voltammetry

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Authors: Sercan Altundemir, Pinar Eribol, A. Kerem Uguz

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Microfluidics systems allow many-large scale laboratory applications to be miniaturized on a single device in order to reduce cost and advance fluid control. Moreover, such systems enable to generate and control droplets which have a significant role on improved analysis for many chemical and biological applications. For example, they can be employed as the model for cells in microfluidic systems. In this work, the interfacial instability of two immiscible Newtonian liquids flowing in a microchannel is investigated. When two immiscible liquids are in laminar regime, a flat interface is formed between them. If a direct current electric field is applied, the interface may deform, i.e. may become unstable and it may be ruptured and form micro-droplets. First, the effect of thickness ratio, total flow rate, viscosity ratio of the silicone oil and ethylene glycol liquid couple on the critical voltage at which the interface starts to destabilize is investigated. Then the droplet sizes are measured under the effect of these parameters at various voltages. Moreover, the effect of total flow rate on the time elapsed for the interface to be ruptured to form droplets by hitting the wall of the channel is analyzed. It is observed that an increase in the viscosity or the thickness ratio of the silicone oil to the ethylene glycol has a stabilizing effect, i.e. a higher voltage is needed while the total flow rate has no effect on it. However, it is observed that an increase in the total flow rate results in shortening of the elapsed time for the interface to hit the wall. Moreover, the droplet size decreases down to 0.1 μL with an increase in the applied voltage, the viscosity ratio or the total flow rate or a decrease in the thickness ratio. In addition to these observations, two empirical models for determining the critical electric number, i.e., the dimensionless voltage and the droplet size and another model which is a combination of both models, for determining the droplet size at the critical voltage are established.

Keywords: droplet formation, electrohydrodynamics, microfluidics, two-phase flow

Procedia PDF Downloads 148