Search results for: ice binding proteins
Commenced in January 2007
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Edition: International
Paper Count: 1676

Search results for: ice binding proteins

356 The Understanding of Biochemical and Molecular Analysis of Diabetic Rats Treated with Andrographis paniculata and Erythrina indica Methanol Extract

Authors: Chakrapani Pullagummi, Arun Jyothi Bheemagani, B. Chandra Sekhar Singh, Prem Kumar, A. Roja Rani

Abstract:

Diabetes mellitus describes a metabolic disorder of multiple aetiology characterized by chronic hyperglycaemia with disturbances of carbohydrate, fat and protein metabolism resulting from defects in insulin secretion and its action. The objective of present study was alloxan induced diabetes in S.D (Sprague Dawley) rats, treated with leaf extract of Andrographis paniculata and bark extract of Erythrina indica. Plant extract treated rats were analyzed biochemically and molecularly. on normal and diabetic rats. The changes in MDA (lipid peroxidation) and glucose (by GOD method) levels in blood of both normal and diabetic rat were analyzed. Diabetes induced rats were treated with methanolic extracts of Andrographis paniculata leaf and Erythrina indica bark which are of medicinal importance. Later after inducing diabetes the rats were treated with medicinal plant extracts, Andrographis paniculata leaf and Erythrina indica bark which are well known for their anti diabetic and antioxidative property in order to control the glucose and MDA levels. The blood plasma of diabetic and normal rats was analyzed for the levels of MDA (lipid peroxidation) and glucose levels. Results of this study suggested that the Andrographis paniculata leaf and Erythrina indica can be used as a potential natural antidiabetic agent for treating and postponing the appearance of complications that arise due to Diabetes. Molecular study deals with the analysis of binding mechanism of 2 selected natural compounds from Andrographis and Erythrina extracts against the novel target for type T2D namely PPAR-γ compared with Rosiglitazone (standard compound). The results revealed that most of the selected herbal lead compounds were effective targets against the receptors. These compounds showed favorable interactions with the amino acid residues thereby substantiating their proven efficacy as anti-diabetic compounds.

Keywords: andrographis paniculata, erythrina indica, alloxan, lipid peroxidation, blood glucose level, PPAR-γ

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355 Characteristics of Smoked Edible Film Made from Myofibril, Collagen and Carrageenan

Authors: Roike Iwan Montolalu, Henny Adeleida Dien, Feny Mentang, Kristhina P. Rahael, Tomy Moga, Ayub Meko, Siegfried Berhimpon

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In the last 20 years, packaging materials derived from petrochemicals polymers were widely used as packaging materials. This due to various advantages such as flexible, strong, transparent, and the price is relatively cheap. However, the plastic polymer also has various disadvantages, such as the transmission monomer contamination into the material to be packed, and waste is non-biodegradable. Edible film (EF) is an up to date materials, generated after the biodegradable packaging materials. The advantages of the EF materials, is the materials can be eat together with food, and the materials can be applied as a coating materials for a widely kind of foods especially snack foods. The aims of this research are to produce and to analyze the characteristics of smoked EF made from carrageenan, myofibril and collagen of Black Marlin (Makaira indica) industrial waste. Smoked EF made with an addition of 0.8 % smoke liquid. Three biopolymers i.e. carrageenan, myofibril, and collagen were used as treatments, and homogenate for 1 hours at speed of 1500 rpm. The analysis carried out on the pH and physical properties i.e. thickness, solubility, tensile strength, % elongation, and water vapor transmission rate (WVTR), as well as on the sensory characteristics of texture i.e. wateriness, firmness, elasticity, hardness, and juiciness of the coated products. The result shown that the higher the concentration the higher the thickness of EF, where as for myofibril proteins appeared higher than carrageenan and collagen. Both of collagen and myofibril shown that concentration of 6% was most soluble, while for carrageenan were in concentration of 2 to 2.5%. For tensile strength, carrageenan was significantly higher than myofibril and collagen; while for elongation, collagen film more elastic than carragenan and myofibril protein. Water vapor transmission rate, shown that myofibril protein film lower than carrageenan and collagen film. From sensory assessment of texture, carrageenan has a high elasticity and juiciness, while collagen and myofibril have a high in firmness and hardness.

Keywords: edible film, collagen, myofibril, carrageenan

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354 Periplasmic Expression of Anti-RoxP Antibody Fragments in Escherichia Coli.

Authors: Caspar S. Carson, Gabriel W. Prather, Nicholas E. Wong, Jeffery R. Anton, William H. McCoy

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Cutibacterium acnes is a commensal bacterium found on human skin that has been linked to acne. C. acnes can also be an opportunistic pathogen when it infiltrates the body during surgery. This pathogen can cause dangerous infections of medical implants, such as shoulder replacements, leading to life-threatening blood infections. Compounding this issue, C. acnes resistance to many antibiotics has become an increasing problem worldwide, creating a need for special forms of treatment. C. acnes expresses the protein RoxP, and it requires this protein to colonize human skin. Though this protein is required for C. acnes skin colonization, its function is not yet understood. Inhibition of RoxP function might be an effective treatment for C. acnes infections. To develop such reagents, the McCoy Laboratory generated four unique anti-RoxP antibodies. Preliminary studies in the McCoy Lab have established that each antibody binds a distinct site on RoxP. To assess the potential of these antibodies as therapeutics, it is necessary to specifically characterize these antibody epitopes and evaluate them in assays that assess their ability to inhibit RoxP-dependent C. acnes growth. To provide material for these studies, an antibody expression construct, Fv-clasp(v2), was adapted to encode anti-RoxP antibody sequences. The author hypothesizes that this expression strategy can produce sufficient amounts of >95% pure antibody fragments for further characterization of these antibodies. Four anti-RoxP Fv-clasp(v2) expression constructs (pET vector-based) were transformed into E. coli BL21-Gold(DE3) cells and a small-scale expression and purification trial was performed for each construct to evaluate anti-RoxP Fv-clasp(v2) yield and purity. Successful expression and purification of these antibody constructs will allow for their use in structural studies, such as protein crystallography and cryogenic electron microscopy. Such studies would help to define the antibody binding sites on RoxP, which could then be leveraged in the development of certain methods to treat C. acnes infection through RoxP inhibition.

Keywords: structural biology, protein expression, infectious disease, antibody, therapeutics, E. coli

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353 Effect of a Muscarinic Antagonist Drug on Extracellular Lipase Activityof Pseudomonas aeruginosa

Authors: Zohreh Bayat, Dariush Minai-Tehrani

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Pseudomonas aeruginosa is a Gram-negative, rode shape and aerobic bacterium that has shown to be resistance to many antibiotics. This resistance makes the bacterium very harmful in some diseases. It can also generate diseases in any part of the gastrointestinal tract from oropharynx to rectum. P. aeruginosa has become an important cause of infection, especially in patients with compromised host defense mechanisms. One of the most important reasons that make P. aeruginosa an emerging opportunistic pathogen in patients is its ability to use various compounds as carbon sources. Lipase is an enzyme that catalyzes the hydrolysis of lipids. Most lipases act at a specific position on the glycerol backbone of lipid substrate. Some lipases are expressed and secreted by pathogenic organisms during the infection. Muscarinic antagonist used as an antispasmodic and in urinary incontinence. The drug has little effect on glandular secretion or the cardiovascular system. It does have some local anesthetic properties and is used in gastrointestinal, biliary, and urinary tract spasms. Aim: In this study the inhibitory effect of a muscarinic antagonist on lipase of P. aeruginosa was investigated. Methods: P. aeruginosa was cultured in minimal salt medium with 1% olive oil as carbon source. The cells were harvested and the supernatant, which contained lipase, was used for enzyme assay. Results: Our results showed that the drug can inhibit P. aeruginosa lipase by competitive manner. In the presence of different concentrations of the drug, the Vmax (2 mmol/min/mg protein) of enzyme did not change, while the Km raised by increasing the drug concentration. The Ki (inhibition constant) and IC50 (the half maximal inhibitory concentration) value of drug was estimated to be about 30 uM and 60 uM which determined that the drug binds to enzyme with high affinity. Maximum activity of the enzyme was observed at pH 8 in the absence and presence of muscarinic antagonist, respectively. The maximum activity of lipase was observed at 600C and the enzyme became inactive at 900C. Conclusion: The muscarinic antagonist drug could inhibit lipase of P. aeruginosa and changed the kinetic parameters of the enzyme. The drug binded to enzyme with high affinity and did not chang the optimum pH of the enzyme. Temperature did not affect the binding of drug to musmuscarinic antagonist.

Keywords: Pseudomonas aeruginosa, drug, enzyme, inhibition

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352 Organic Carbon Pools Fractionation of Lacustrine Sediment with a Stepwise Chemical Procedure

Authors: Xiaoqing Liu, Kurt Friese, Karsten Rinke

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Lacustrine sediment archives rich paleoenvironmental information in lake and surrounding environment. Additionally, modern sediment is used as an effective medium for the monitoring of lake. Organic carbon in sediment is a heterogeneous mixture with varying turnover times and qualities which result from the different biogeochemical processes in the deposition of organic material. Therefore, the isolation of different carbon pools is important for the research of lacustrine condition in the lake. However, the numeric available fractionation procedures can hardly yield homogeneous carbon pools on terms of stability and age. In this work, a multi-step fractionation protocol that treated sediment with hot water, HCl, H2O2 and Na2S2O8 in sequence was adopted, the treated sediment from each step were analyzed for the isotopic and structural compositions with Isotope Ratio Mass Spectrometer coupled with element analyzer (IRMS-EA) and Solid-state 13C Nuclear Magnetic Resonance (NMR), respectively. The sequential extractions with hot-water, HCl, and H2O2 yielded a more homogeneous and C3 plant-originating OC fraction, which was characterized with an atomic C/N ratio shift from 12.0 to 20.8, and 13C and 15N isotopic signatures were 0.9‰ and 1.9‰ more depleted than the original bulk sediment, respectively. Additionally, the H2O2- resistant residue was dominated with stable components, such as the lignins, waxes, cutans, tannins, steroids and aliphatic proteins and complex carbohydrates. 6M HCl in the acid hydrolysis step was much more effective than 1M HCl to isolate a sedimentary OC fraction with higher degree of homogeneity. Owing to the extremely high removal rate of organic matter, the step of a Na2S2O8 oxidation is only suggested if the isolation of the most refractory OC pool is mandatory. We conclude that this multi-step chemical fractionation procedure is effective to isolate more homogeneous OC pools in terms of stability and functional structure, and it can be used as a promising method for OC pools fractionation of sediment or soil in future lake research.

Keywords: 13C-CPMAS-NMR, 13C signature, lake sediment, OC fractionation

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351 Colour and Travel: Design of an Innovative Infrastructure for Travel Applications with Entertaining and Playful Features

Authors: Avrokomi Zavitsanou, Spiros Papadopoulos, Theofanis Alexandridis

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This paper presents the research project ‘Colour & Travel’, which is co-funded by the European Union and national resources through the Operational Programme “Competitiveness, Entrepreneurship and Innovation” 2014-2020, under the Single RTDI State Aid Action "RESEARCH - CREATE - INNOVATE". The research project proposes the design of an innovative, playful framework for exploring a variety of travel destinations and creating personalised travel narratives, aiming to entertain, educate, and promote culture and tourism. Gamification of the cultural and touristic environment can enhance its experiential, multi-sensory aspects and broaden the perception of the traveler. The latter's involvement in creating and shaping his personal travel narrations and the possibility of sharing it with others can offer him an alternative, more binding way of getting acquainted with a place. In particular, the paper presents the design of an infrastructure: (a) for the development of interactive travel guides for mobile devices, where sites with specific points of interest will be recommended, with which the user can interact in playful ways and then create his personal travel narratives, (b) for the development of innovative games within virtual reality environment, where the interaction will be offered while the user is moving within the virtual environment; and (c) for an online application where the content will be offered through the browser and the modern 3D imaging technologies (WebGL). The technological products that will be developed within the proposed project can strengthen important sectors of economic and social life, such as trade, tourism, exploitation and promotion of the cultural environment, creative industries, etc. The final applications delivered at the end of the project will guarantee an improved level of service for visitors and will be a useful tool for content creators with increased adaptability, expansibility, and applicability in many regions of Greece and abroad. This paper aims to present the research project by referencing the state of the art and the methodological scheme, ending with a brief reflection on the expected outcome in terms of results.

Keywords: gamification, culture, tourism, AR, VR, applications

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350 The Examination And Assurance Of The Microbiological Safety Pertaining To Raw Milk And its Derived Processed Products

Authors: Raana Babadi Fathipour

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The production of dairy holds significant importance in the sustenance of billions of individuals worldwide, as they rely on milk and its derived products for daily consumption. In addition to being a source of essential nutrients crucial for human well-being, such as proteins, fats, vitamins, and minerals; dairy items are witnessing an increasing demand worldwide. Amongst all the factors contributing to the quality and safety assurance of dairy products, the strong focus lies on maintaining high standards in raw milk procurement. Raw milk serves as an externally nutritious medium for various microorganisms due to its inherent properties. This poses a considerable challenge for the dairy industry in ensuring that microbial contamination is minimized throughout every stage of the value chain. Despite implementing diverse process technologies—both conventional and innovative—the occurrence of microbial spoilage still results in substantial losses within this industry context. Moreover, milk and dairy products have been associated with numerous cases of foodborne illnesses across the globe. Various pathogens such as Salmonella serovars, Campylobacter spp., Shiga toxin-producing Escherichia coli, Listeria monocytogenes, and enterotoxin producing Staphylococcus aureus are commonly identified as the culprits behind these outbreaks in the dairy industry. The effective management of food safety within this sector necessitates a proactive and risk-based approach to reform. However, this strategy presents difficulties for developing nations where informal value chains dominate the dairy sector. Whether operating on a small or large scale or falling within formal or informal realms, it is imperative that the dairy industry adheres to principles of good hygiene practices and good manufacturing practices. Additionally, identifying and managing potential sources of contamination is crucial in mitigating challenges pertaining to quality and safety precautions.

Keywords: dairy value chain, microbial contamination, food safety, hygiene

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349 Bioinformatic Design of a Non-toxic Modified Adjuvant from the Native A1 Structure of Cholera Toxin with Membrane Synthetic Peptide of Naegleria fowleri

Authors: Frida Carrillo Morales, Maria Maricela Carrasco Yépez, Saúl Rojas Hernández

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Naegleria fowleri is the causative agent of primary amebic meningoencephalitis, this disease is acute and fulminant that affects humans. It has been reported that despite the existence of therapeutic options against this disease, its mortality rate is 97%. Therefore, the need arises to have vaccines that confer protection against this disease and, in addition to developing adjuvants to enhance the immune response. In this regard, in our work group, we obtained a peptide designed from the membrane protein MP2CL5 of Naegleria fowleri called Smp145 that was shown to be immunogenic; however, it would be of great importance to enhance its immunological response, being able to co-administer it with a non-toxic adjuvant. Therefore, the objective of this work was to carry out the bioinformatic design of a peptide of the Naegleria fowleri membrane protein MP2CL5 conjugated with a non-toxic modified adjuvant from the native A1 structure of Cholera Toxin. For which different bioinformatics tools were used to obtain a model with a modification in amino acid 61 of the A1 subunit of the CT (CTA1), to which the Smp145 peptide was added and both molecules were joined with a 13-glycine linker. As for the results obtained, the modification in CTA1 bound to the peptide produces a reduction in the toxicity of the molecule in in silico experiments, likewise, the prediction in the binding of Smp145 to the receptor of B cells suggests that the molecule is directed in specifically to the BCR receptor, decreasing its native enzymatic activity. The stereochemical evaluation showed that the generated model has a high number of adequately predicted residues. In the ERRAT test, the confidence with which it is possible to reject regions that exceed the error values was evaluated, in the generated model, a high score was obtained, which determines that the model has a good structural resolution. Therefore, the design of the conjugated peptide in this work will allow us to proceed with its chemical synthesis and subsequently be able to use it in the mouse meningitis protection model caused by N. fowleri.

Keywords: immunology, vaccines, pathogens, infectious disease

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348 Developing Novel Bacterial Primase (DnaG) Inhibitors

Authors: Shanakr Bhattarai, V. S. Tiwari, Barak Akabayov

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The plummeting number of infections and death is due to the development of drug-resistant bacteria. In addition, the number of approved antibiotic drugs by the Food and Drug Administration (FDA) is insufficient. Therefore, developing new drugs and finding novel targets for central metabolic pathways in bacteria is urgently needed. One of the promising targets is DNA replication machinery which consists of many essential proteins and enzymes. DnaG primase is an essential enzyme and a central part of the DNA replication machinery. DnaG primase synthesizes short RNA primers that initiate the Okazaki fragments by the lagging strand DNA polymerase. Therefore, it is reasonable to assume that inhibition of primase activity will stall DNA replication and prevent bacterial proliferation. We did the expression and purification of eight different bacterial DnaGs (Mycobacterium tuberculosis(Mtb), Bacillus anthracis (Ba), Mycobacterium smegmatis (Msmeg), Francisella tularencis (Ft), Vibrio cholerae (Vc) and Yersinia pestis (Yp), Staphylococcus aureus(Saureus), Escherichia coli(Ecoli)) followed by the radioactive activity assay. After obtaining the pure and active protein DnaG, we synthesized the inhibitors for them. The inhibitors were divided into five different groups, each containing five molecules, and the cocktail inhibition assay was performed against each DnaGs. The groups of molecules inhibiting the DnaGs were further tested with individual molecules belonging to inhibiting groups. Each molecule showing inhibition was titrated against the corresponding DnaGs to find IC50. We got a molecule(VS167) that acted as broad inhibitors, inhibiting all eight DnaGs. Molecules VS180 and VS186 inhibited seven DnaGs (except Saureus). Similarly, two molecules(VS 173, VS176) inhibited five DnaGs (Mtb, Ba, Ft, Yp, Ecoli). VS261 inhibited four DnaGs (Mtb, Ba, Ft, Vc). MS50 inhibited Ba and Vc DnaGs. And some of the inhibitors inhibited only one DnaGs. Thus we found the broad and specific inhibitors for different bacterial DnaGs, and their Structure-activity analysis(SAR) was done. Further, We tried to explain the similarities among the enzyme DnaGs from different bacteria based on their inhibition pattern.

Keywords: DNA replication, DnaG, okazaki fragments, antibiotic drugs

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347 Three Issues for Integrating Artificial Intelligence into Legal Reasoning

Authors: Fausto Morais

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Artificial intelligence has been widely used in law. Programs are able to classify suits, to identify decision-making patterns, to predict outcomes, and to formalize legal arguments as well. In Brazil, the artificial intelligence victor has been classifying cases to supreme court’s standards. When those programs act doing those tasks, they simulate some kind of legal decision and legal arguments, raising doubts about how artificial intelligence can be integrated into legal reasoning. Taking this into account, the following three issues are identified; the problem of hypernormatization, the argument of legal anthropocentrism, and the artificial legal principles. Hypernormatization can be seen in the Brazilian legal context in the Supreme Court’s usage of the Victor program. This program generated efficiency and consistency. On the other hand, there is a feasible risk of over standardizing factual and normative legal features. Then legal clerks and programmers should work together to develop an adequate way to model legal language into computational code. If this is possible, intelligent programs may enact legal decisions in easy cases automatically cases, and, in this picture, the legal anthropocentrism argument takes place. Such an argument argues that just humans beings should enact legal decisions. This is so because human beings have a conscience, free will, and self unity. In spite of that, it is possible to argue against the anthropocentrism argument and to show how intelligent programs may work overcoming human beings' problems like misleading cognition, emotions, and lack of memory. In this way, intelligent machines could be able to pass legal decisions automatically by classification, as Victor in Brazil does, because they are binding by legal patterns and should not deviate from them. Notwithstanding, artificial intelligent programs can be helpful beyond easy cases. In hard cases, they are able to identify legal standards and legal arguments by using machine learning. For that, a dataset of legal decisions regarding a particular matter must be available, which is a reality in Brazilian Judiciary. Doing such procedure, artificial intelligent programs can support a human decision in hard cases, providing legal standards and arguments based on empirical evidence. Those legal features claim an argumentative weight in legal reasoning and should serve as references for judges when they must decide to maintain or overcome a legal standard.

Keywords: artificial intelligence, artificial legal principles, hypernormatization, legal anthropocentrism argument, legal reasoning

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346 Pale, Soft, Exudative (PSE) Turkey Meat in a Brazilian Commercial Processing Plant

Authors: Danielle C. B. Honorato, Rafael H. Carvalho, Adriana L. Soares, Ana Paula F. R. L. Bracarense, Paulo D. Guarnieri, Massami Shimokomaki, Elza I. Ida

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Over the past decade, the Brazilian production of turkey meat increased by more than 50%, indicating that the turkey meat is considered a great potential for the Brazilian economy contributing to the growth of agribusiness at the marketing international scenario. However, significant color changes may occur during its processing leading to the pale, soft and exudative (PSE) appearance on the surface of breast meat due to the low water holding capacity (WHC). Changes in PSE meat functional properties occur due to the myofibrils proteins denaturation caused by a rapid postmortem glycolysis resulting in a rapid pH decline while the carcass temperature is still warm. The aim of this study was to analyze the physical, chemical and histological characteristics of PSE turkey meat obtained from a Brazilian commercial processing plant. The turkey breasts samples were collected (n=64) at the processing line and classified as PSE at L* ≥ 53 value. The pH was also analyzed after L* measurement. In sequence, PSE meat samples were evaluated for WHC, cooking loss (CL), shear force (SF), myofibril fragmentation index (MFI), protein denaturation (PD) and histological evaluation. The abnormal color samples presented lower pH values, 16% lower fiber diameter, 11% lower SF and 2% lower WHC than those classified as normal. The CL, PD and MFI were, respectively, 9%, 18% and 4% higher in PSE samples. The Pearson correlation between the L* values and CL, PD and MFI was positive, while that SF and pH values presented negative correlation. Under light microscopy, a shrinking of PSE muscle cell diameter was approximately 16% shorter in relation to normal samples and an extracellular enlargement of endomysium and perimysium sheaths as the consequence of higher water contents lost as observed previously by lower WHC values. Thus, the results showed that PSE turkey breast meat presented significant changes in their physical, chemical and histological characteristics that may impair its functional properties.

Keywords: functional properties, histological evaluation, meat quality, PSE

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345 Improved Non-Ideal Effects in AlGaN/GaN-Based Ion-Sensitive Field-Effect Transistors

Authors: Wei-Chou Hsu, Ching-Sung Lee, Han-Yin Liu

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This work uses H2O2 oxidation technique to improve the pH sensitivity of the AlGaN/GaN-based ion-sensitive field-effect transistors (ISFETs). 10-nm-thick Al2O3 was grown on the surface of the AlGaN. It was found that the pH sensitivity was improved from 41.6 mV/pH to 55.2 mV/pH. Since the H2O2-grown Al2O3 was served as a passivation layer and the problem of Fermi-level pinning was suppressed for the ISFET with the H2O2 oxidation process. Hysteresis effect in the ISFET with the H2O2 treatment also became insignificant. The hysteresis effect was observed by dipping the ISFETs into different pH value solutions and comparing the voltage difference between the initial and final conditions. The hysteresis voltage (Vhys) of the ISFET with the H2O2 oxidation process was improved from 8.7 mV to 4.8 mV. The hysteresis effect is related to the buried binding sites which are related to the material defects like threading dislocations in the AlGaN/GaN heterostructure which was grown by the hetero-epitaxy technique. The H2O2-grown Al2O3 passivate these material defects and the Al2O3 has less material defects. The long-term stability of the ISFET is estimated by the drift effect measurement. The drift measurement was conducted by dipping the ISFETs into a specific pH value solution for 12 hours and the ISFETs were operating at a specific quiescent point. The drift rate is estimated by the drift voltage divided by the total measuring time. It was found that the drift rate of the ISFET was improved from 10.1 mV/hour to 1.91 mV/hour in the pH 7 solution, from 14.06 mV/hour to 6.38 mV/pH in the pH 2 solution, and from 12.8 mV/hour to 5.48 mV/hour in the pH 12 solution. The drift effect results from the capacitance variation in the electric double layer. The H2O2-grown Al2O3 provides an additional capacitance connection in series with the electric double layer. Therefore, the capacitance variation of the electric double layer became insignificant. Generally, the H2O2 oxidation process is a simple, fast, and cost-effective method for the AlGaN/GaN-based ISFET. Furthermore, the performance of the AlGaN/GaN ISFET was improved effectively and the non-ideal effects were suppressed.

Keywords: AlGaN/GaN, Al2O3, hysteresis effect, drift effect, reliability, passivation, pH sensors

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344 Rapid and Cheap Test for Detection of Streptococcus pyogenes and Streptococcus pneumoniae with Antibiotic Resistance Identification

Authors: Marta Skwarecka, Patrycja Bloch, Rafal Walkusz, Oliwia Urbanowicz, Grzegorz Zielinski, Sabina Zoledowska, Dawid Nidzworski

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Upper respiratory tract infections are one of the most common reasons for visiting a general doctor. Streptococci are the most common bacterial etiological factors in these infections. There are many different types of Streptococci and infections vary in severity from mild throat infections to pneumonia. For example, S. pyogenes mainly contributes to acute pharyngitis, palatine tonsils and scarlet fever, whereas S. Streptococcus pneumoniae is responsible for several invasive diseases like sepsis, meningitis or pneumonia with high mortality and dangerous complications. There are only a few diagnostic tests designed for detection Streptococci from the infected throat of patients. However, they are mostly based on lateral flow techniques, and they are not used as a standard due to their low sensitivity. The diagnostic standard is to culture patients throat swab on semi selective media in order to multiply pure etiological agent of infection and subsequently to perform antibiogram, which takes several days from the patients visit in the clinic. Therefore, the aim of our studies is to develop and implement to the market a Point of Care device for the rapid identification of Streptococcus pyogenes and Streptococcus pneumoniae with simultaneous identification of antibiotic resistance genes. In the course of our research, we successfully selected genes for to-species identification of Streptococci and genes encoding antibiotic resistance proteins. We have developed a reaction to amplify these genes, which allows detecting the presence of S. pyogenes or S. pneumoniae followed by testing their resistance to erythromycin, chloramphenicol and tetracycline. What is more, the detection of β-lactamase-encoding genes that could protect Streptococci against antibiotics from the ampicillin group, which are widely used in the treatment of this type of infection is also developed. The test is carried out directly from the patients' swab, and the results are available after 20 to 30 minutes after sample subjection, which could be performed during the medical visit.

Keywords: antibiotic resistance, Streptococci, respiratory infections, diagnostic test

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343 Lead Chalcogenide Quantum Dots for Use in Radiation Detectors

Authors: Tom Nakotte, Hongmei Luo

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Lead chalcogenide-based (PbS, PbSe, and PbTe) quantum dots (QDs) were synthesized for the purpose of implementing them in radiation detectors. Pb based materials have long been of interest for gamma and x-ray detection due to its high absorption cross section and Z number. The emphasis of the studies was on exploring how to control charge carrier transport within thin films containing the QDs. The properties of QDs itself can be altered by changing the size, shape, composition, and surface chemistry of the dots, while the properties of carrier transport within QD films are affected by post-deposition treatment of the films. The QDs were synthesized using colloidal synthesis methods and films were grown using multiple film coating techniques, such as spin coating and doctor blading. Current QD radiation detectors are based on the QD acting as fluorophores in a scintillation detector. Here the viability of using QDs in solid-state radiation detectors, for which the incident detectable radiation causes a direct electronic response within the QD film is explored. Achieving high sensitivity and accurate energy quantification in QD radiation detectors requires a large carrier mobility and diffusion lengths in the QD films. Pb chalcogenides-based QDs were synthesized with both traditional oleic acid ligands as well as more weakly binding oleylamine ligands, allowing for in-solution ligand exchange making the deposition of thick films in a single step possible. The PbS and PbSe QDs showed better air stability than PbTe. After precipitation the QDs passivated with the shorter ligand are dispersed in 2,6-difloupyridine resulting in colloidal solutions with concentrations anywhere from 10-100 mg/mL for film processing applications, More concentrated colloidal solutions produce thicker films during spin-coating, while an extremely concentrated solution (100 mg/mL) can be used to produce several micrometer thick films using doctor blading. Film thicknesses of micrometer or even millimeters are needed for radiation detector for high-energy gamma rays, which are of interest for astrophysics or nuclear security, in order to provide sufficient stopping power.

Keywords: colloidal synthesis, lead chalcogenide, radiation detectors, quantum dots

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342 Collagen Hydrogels Cross-Linked by Squaric Acid

Authors: Joanna Skopinska-Wisniewska, Anna Bajek, Marta Ziegler-Borowska, Alina Sionkowska

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Hydrogels are a class of materials widely used in medicine for many years. Proteins, such as collagen, due to the presence of a large number of functional groups are easily wettable by polar solvents and can create hydrogels. The supramolecular network capable to swelling is created by cross-linking of the biopolymers using various reagents. Many cross-linking agents has been tested for last years, however, researchers still are looking for a new, more secure reactants. Squaric acid, 3,4-dihydroxy 3-cyclobutene 1,2- dione, is a very strong acid, which possess flat and rigid structure. Due to the presence of two carboxyl groups the squaric acid willingly reacts with amino groups of collagen. The main purpose of this study was to investigate the influence of addition of squaric acid on the chemical, physical and biological properties of collagen materials. The collagen type I was extracted from rat tail tendons and 1% solution in 0.1M acetic acid was prepared. The samples were cross-linked by the addition of 5%, 10% and 20% of squaric acid. The mixtures of all reagents were incubated 30 min on magnetic stirrer and then dialyzed against deionized water. The FTIR spectra show that the collagen structure is not changed by cross-linking by squaric acid. Although the mechanical properties of the collagen material deteriorate, the temperature of thermal denaturation of collagen increases after cross-linking, what indicates that the protein network was created. The lyophilized collagen gels exhibit porous structure and the pore size decreases with the higher addition of squaric acid. Also the swelling ability is lower after the cross-linking. The in vitro study demonstrates that the materials are attractive for 3T3 cells. The addition of squaric acid causes formation of cross-ling bonds in the collagen materials and the transparent, stiff hydrogels are obtained. The changes of physicochemical properties of the material are typical for cross-linking process, except mechanical properties – it requires further experiments. However, the results let us to conclude that squaric acid is a suitable cross-linker for protein materials for medicine and tissue engineering.

Keywords: collagen, squaric acid, cross-linking, hydrogel

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341 Comparison of Inexpensive Cell Disruption Techniques for an Oleaginous Yeast

Authors: Scott Nielsen, Luca Longanesi, Chris Chuck

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Palm oil is obtained from the flesh and kernel of the fruit of oil palms and is the most productive and inexpensive oil crop. The global demand for palm oil is approximately 75 million metric tonnes, a 29% increase in global production of palm oil since 2016. This expansion of oil palm cultivation has resulted in mass deforestation, vast biodiversity destruction and increasing net greenhouse gas emissions. One possible alternative is to produce a saturated oil, similar to palm, from microbes such as oleaginous yeast. The yeasts can be cultured on sugars derived from second-generation sources and do not compete with tropical forests for land. One highly promising oleaginous yeast for this application is Metschnikowia pulcherrima. However, recent techno-economic modeling has shown that cell lysis and standard lipid extraction are major contributors to the cost of the oil. Typical cell disruption techniques to extract either single cell oils or proteins have been based around bead-beating, homogenization and acid lysis. However, these can have a detrimental effect on lipid quality and are energy-intensive. In this study, a vortex separator, which produces high sheer with minimal energy input, was investigated as a potential low energy method of lysing cells. This was compared to four more traditional methods (thermal lysis, acid lysis, alkaline lysis, and osmotic lysis). For each method, the yeast loading was also examined at 1 g/L, 10 g/L and 100 g/L. The quality of the cell disruption was measured by optical cell density, cell counting and the particle size distribution profile comparison over a 2-hour period. This study demonstrates that the vortex separator is highly effective at lysing the cells and could potentially be used as a simple apparatus for lipid recovery in an oleaginous yeast process. The further development of this technology could potentially reduce the overall cost of microbial lipids in the future.

Keywords: palm oil substitute, metschnikowia pulcherrima, cell disruption, cell lysis

Procedia PDF Downloads 182
340 Effect of Diazepam on Internal Organs of Chrysomya megacephala Using Micro-Computed Tomograph

Authors: Sangkhao M., Butcher B. A.

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Diazepam (known as valium) is a medication for calming effect. Many reports on committed suicide cases shown that diazepam is frequently used for this purpose. This research aims to study effect of diazepam on the development of forensically important blowflies, Chrysomya megacephala (Diptera: Calliphoridae) using micro-computed tomography (micro CT). In this study, four rabbits were treated with three different lethal doses of diazepam and one control (LD₀, LD₅₀, LD₁₀₀ and LC). The rabbit’s livers were removed for rearing the blowflies. Pupae were sampled for two series (ages; S1: 24h and S2: 120h) of development. After preparing the specimens, all samples were performed Micro CT using Skyscan 1172. The results shown the effect of diazepam on internal organs and tissues such as brain, cavity of the body, gas bubble, meconium and especially fat body. In the control group, in series 1 (LCS1), fat body was equally dispersed in the head, thorax, and abdomen, development of internal organs were not completed, however, brain, thoracic muscle, wings, legs and rectum were able to observe at 24h after developing into the pupal stage. Development of each organ in the control group in the series two was completed. In the treatment groups, LD₀, LD₅₀, LD₁₀₀ (Series 1 and Series 2), tissues are different, such as gas bubble in LD₀S1, was observed due to rapidity morphological changes during the metamorphosis of blowfly’s pupa in this treatment. Meconium was observed in LD₅₀S2 group because excretion of metabolic waste was not completed. All of the samples in the treatment groups had differentiation of fat bodies because metabolic activities were not completed and these changes affected on functions of every internal system. Discovering of differentiated fat bodies are important results because fat bodies of insect functions as liver in human, therefore it is shown that toxin eliminates from blowfly’s body and homeostatic maintenance of the hemolymph proteins, lipid and carbohydrates in each treatment group are abnormal.

Keywords: forensic toxicology, forensic entomology, diptera, diazepam

Procedia PDF Downloads 118
339 Transcriptome Analysis of Saffron (crocus sativus L.) Stigma Focusing on Identification Genes Involved in the Biosynthesis of Crocin

Authors: Parvaneh Mahmoudi, Ahmad Moeni, Seyed Mojtaba Khayam Nekoei, Mohsen Mardi, Mehrshad Zeinolabedini, Ghasem Hosseini Salekdeh

Abstract:

Saffron (Crocus sativus L.) is one of the most important spice and medicinal plants. The three-branch style of C. sativus flowers are the most important economic part of the plant and known as saffron, which has several medicinal properties. Despite the economic and biological significance of this plant, knowledge about its molecular characteristics is very limited. In the present study, we, for the first time, constructed a comprehensive dataset for C. sativus stigma through de novo transcriptome sequencing. We performed de novo transcriptome sequencing of C. sativus stigma using the Illumina paired-end sequencing technology. A total of 52075128 reads were generated and assembled into 118075 unigenes, with an average length of 629 bp and an N50 of 951 bp. A total of 66171unigenes were identified, among them, 66171 (56%) were annotated in the non-redundant National Center for Biotechnology Information (NCBI) database, 30938 (26%) were annotated in the Swiss-Prot database, 10273 (8.7%) unigenes were mapped to 141 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database, while 52560 (44%) and 40756 (34%) unigenes were assigned to Gen Ontology (GO) categories and Eukaryotic Orthologous Groups of proteins (KOG), respectively. In addition, 65 candidate genes involved in three stages of crocin biosynthesis were identified. Finally, transcriptome sequencing of saffron stigma was used to identify 6779 potential microsatellites (SSRs) molecular markers. High-throughput de novo transcriptome sequencing provided a valuable resource of transcript sequences of C. sativus in public databases. In addition, most of candidate genes potentially involved in crocin biosynthesis were identified which could be further utilized in functional genomics studies. Furthermore, numerous obtained SSRs might contribute to address open questions about the origin of this amphiploid spices with probable little genetic diversity.

Keywords: saffron, transcriptome, NGS, bioinformatic

Procedia PDF Downloads 76
338 Heavy Metals and Carcinogenic Risk Assessment in Free-Ranged Livestock of Lead-Contaminated Goldmine Communities of Zamfara State, Northern Nigeria

Authors: Sulaiman Rabiu, Muazu Gusau Abubakar, Jafar Usman Zakari

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The consumption of meat is of great importance as it provides a good source of proteins and significant amount of essential trace element to the body. However, contamination of meat and meat products with heavy metals is becoming a serious threat to food safety and public health. Therefore, the present study is aimed to evaluate the concentration of some heavy metals in muscles and entrails of free-ranged cattle, sheep and goats. A total of sixty (60) fresh samples of muscles, liver, kidney, small intestines and stomach of free ranged cattle, sheep and goats were collected from abattoirs of different goldmine communities of Anka, Bukkuyum, Maru andTalata-Mafara Local Government Areas of Zamfara State, Nigeria. The samples were digested using 10 mL of a mixed 70% high grade concentration of HNO₃ and 65% HCl (4:1 v/v); the mixture was heated until dense fumes disappeared forming a clear transparent solution and diluted to 50 mL with deionized water. Actual concentrations of Cd, Cr, Cu, Co, As, Ni, Mn, Pb and Zn were determined using Microwave Plasma Atomic Emission Spectrophotometer (MP-AES). From the results obtained, goat liver had the highest mean concentration of lead, arsenic, cobalt and manganese (12.43± 0.31, 14.25±0.32, 3.47± 0.86 and 12.68± 0.92 mg/kg respectively) while goat kidney had the highest concentration of copper and zinc (10.08±0.61 and 24.16±1.30 mg/kg respectively). The highest concentrations of cadmium and nickel were recorded in sheep kidney (7.75± 0.65 and 2.08±0.10 mg/kg respectively). Cattle muscles had the highest chromium concentration than all the organs analysed. The target hazard quotients (THQs) for all the metals were below 1.0, but TR which is a risk indices for carcinogenicity indicates an alarming result that requires stringent control to protect public health.Therefore, intensive public health awareness on the risk associated with contamination of heavy metals in meat should be advocated.

Keywords: contamination, goldmine, heavy metals, meat

Procedia PDF Downloads 74
337 Construction of Genetic Recombinant Yeasts with High Environmental Tolerance by Accumulation of Trehalose and Detoxication of Aldehyde

Authors: Yun-Chin Chung, Nileema Divate, Gen-Hung Chen, Pei-Ru Huang, Rupesh Divate

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Many environmental factors, such as glucose concentration, ethanol, temperature, osmotic pressure and pH, decrease the production rate of ethanol using yeast as a starter. Fermentation starters with high tolerance to various stresses are always demanded for brewing industry. Trehalose, a storage carbohydrate in cell wall of yeast, plays an important role in tolerance of environmental stress by preserving integrity of plasma membrane and stabilizing proteins. Furan aldehydes are toxic to yeast and the growth rate of yeast is significantly reduced if furan aldehydes were present in the fermentation medium. In yeast, aldehyde reductase is involved in the detoxification of reactive aldehydes and consequently the growth of yeast is improved. The aims of this study were to construct a genetic recombinant Saccharomyces cerevisiae or Pichia pastoris with furfural and HMF degrading and high ethanol tolerance capacities. Yeast strains were engineered by genetic recombination for overexpression of trehalose-6-phosphate synthase gene (tps1) and aldehyde reductase gene (ari1). TPS1 gene was cloned from S. cerevisiae by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated with pGAPZαC vector. The constructed vector, pGAPZC-tps1, was transformed to recombinant yeasts strain with overexpression of ari1. The transformants with pGAPZC-tps1-ari1 were generated called STA (S. cerevisiae) and PTA (P. pastoris) with overexpression of tps1, ari1. PCR with tps1-specific primers and western blot with his-tag confirmed the gene insertion and protein expression of tps1 in the transformants, respectively. The neutral trehalase gene (nth1) of STA was successfully deleted and the novel strain STAΔN will be used for further study, including the measurement of trehalose concentration and ethanol, furfural tolerance assay.

Keywords: genetic recombinant, yeast, ethanol tolerance, trehalase, aldehyde reductase

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336 Therapeutic Efficacy of Clompanus Pubescens Leaves Fractions via Downregulation of Neuronal Cholinesterases/NA⁺-K⁺ ATPase/IL-1 β and Improving the Neurocognitive and Antioxidants Status of Streptozotocin-Induced Diabetic Rats

Authors: Amos Sunday Onikanni, Bashir Lawal, Babatunji Emmanuel Oyinloye, Gomaa Mostafa-Hedeab, Mohammed Alorabi, Simona Cavalu, Augustine O. Olusola, Chih-Hao Wang, Gaber El-Saber Batiha

Abstract:

The increasing global burden of diabetes mellitus has called for the search for a therapeutic alternative that offers better activities and safety than conventional chemotherapy. Herein, we evaluated the neuroprotective and antioxidant properties of different fractions (ethyl acetate, N-butanol and residual aqueous) of Clompanus pubescens leaves in streptozotocin (STZ)-induced diabetic rats. Our results revealed a significant elevation in the levels of blood glucose, pro-inflammatory cytokines, lipid peroxidation, neuronal activities of acetylcholinesterase, butyrylcholinesterase, nitric oxide, epinephrine, norepinephrine, and Na+/K+-ATPase in diabetic non treated rats. In addition, decreased levels of enzymatic and non-enzymatic antioxidants were observed. Treatment with different fractions of C. pubescens leaves resulted in a significant reversal of the biochemical alteration and improved the neurocognitive deficit in STZ-induced diabetic rats. However, the ethyl-acetate fraction demonstrated higher activities than the other fractions and was characterized for its phytoconstituents, revealing the presence of Gallic acid (713.00 ppm), catechin (0.91 ppm), ferulic acid (0.98 ppm), rutin (59.82 ppm), quercetin (3.22 ppm) and kaempferol (4.07 ppm). Our molecular docking analysis revealed that these compounds exhibited different binding affinities and potentials for targeting BChE/AChE/ IL-1 β/Na+-K+-ATPase. However, only Kampferol and ferulic exhibited good drug-like, ADMET, and permeability properties suitable for use as a neuronal drug target agent. Hence, the ethyl-acetate fraction of C. pubescent leaves could be considered a source of promising bioactive metabolite for the treatment and management of cognitive impairments related to type II diabetes mellitus.

Keywords: diabetes mellitus, neuroprotective, antioxidant, pro-inflammatory cytokines

Procedia PDF Downloads 98
335 Wet Processing of Algae for Protein and Carbohydrate Recovery as Co-Product of Algal Oil

Authors: Sahil Kumar, Rajaram Ghadge, Ramesh Bhujade

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Historically, lipid extraction from dried algal biomass remained a focus area of the algal research. It has been realized over the past few years that the lipid-centric approach and conversion technologies that require dry algal biomass have several challenges. Algal culture in cultivation systems contains more than 99% water, with algal concentrations of just a few hundred milligrams per liter ( < 0.05 wt%), which makes harvesting and drying energy intensive. Drying the algal biomass followed by extraction also entails the loss of water and nutrients. In view of these challenges, focus has shifted toward developing processes that will enable oil production from wet algal biomass without drying. Hydrothermal liquefaction (HTL), an emerging technology, is a thermo-chemical conversion process that converts wet biomass to oil and gas using water as a solvent at high temperature and high pressure. HTL processes wet algal slurry containing more than 80% water and significantly reduces the adverse cost impact owing to drying the algal biomass. HTL, being inherently feedstock agnostic, i.e., can convert carbohydrates and proteins also to fuels and recovers water and nutrients. It is most effective with low-lipid (10--30%) algal biomass, and bio-crude yield is two to four times higher than the lipid content in the feedstock. In the early 2010s, research remained focused on increasing the oil yield by optimizing the process conditions of HTL. However, various techno-economic studies showed that simply converting algal biomass to only oil does not make economic sense, particularly in view of low crude oil prices. Making the best use of every component of algae is a key for economic viability of algal to oil process. On investigation of HTL reactions at the molecular level, it has been observed that sequential HTL has the potential to recover value-added products along with biocrude and improve the overall economics of the process. This potential of sequential HTL makes it a most promising technology for converting wet waste to wealth. In this presentation, we will share our experience on the techno-economic and engineering aspects of sequential HTL for conversion of algal biomass to algal bio-oil and co-products.

Keywords: algae, biomass, lipid, protein

Procedia PDF Downloads 192
334 Molecular Characterisation and Expression of Glutathione S-Transferase of Fasciola Gigantica

Authors: J. Adeppa, S. Samanta, O. K. Raina

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Fasciolosis is a widespread economically important parasitic infection throughout the world caused by Fasciola hepatica and F. gigantica. In order to identify novel immunogen conferring significant protection against fasciolosis, currently, research has been focused on the defined antigens viz. glutathione S-transferase, fatty acid binding protein, cathepsin-L, fluke hemoglobin, paramyosin, myosin and F. hepatica- Kunitz Type Molecule. Among various antigens, GST which plays a crucial role in detoxification processes, i.e. phase II defense mechanism of this parasite, has a unique position as a novel vaccine candidate and a drug target in the control of this disease. For producing the antigens in large quantities and their purification to complete homogeneity, the recombinant DNA technology has become an important tool to achieve this milestone. RT- PCR was carried out using F. gigantica total RNA as template, and an amplicon of 657 bp GST gene was obtained. TA cloning vector was used for cloning of this gene, and the presence of insert was confirmed by blue-white selection for recombinant colonies. Sequence analysis of the present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin (accession no. AF112657), with six nucleotide changes at 72, 74, 423, 513, 549 and 627th bp found in the present isolate, causing an overall change of 4 amino acids. The 657 bp GST gene was cloned at BamH1 and HindIII restriction sites of the prokaryotic expression vector pPROEXHTb in frame with six histidine residues and expressed in E. coli DH5α. Recombinant protein was purified from the bacterial lysate under non-denaturing conditions by the process of sonication after lysozyme treatment and subjecting the soluble fraction of the bacterial lysate to Ni-NTA affinity chromatography. Western blotting with rabbit hyper-immune serum showed immuno-reactivity with 25 kDa recombinant GST. Recombinant protein detected F. gigantica experimental as well as field infection in buffaloes by dot-ELISA. However, cross-reactivity studies on Fasciola gigantica GST antigen are needed to evaluate the utility of this protein in the serodiagnosis of fasciolosis.

Keywords: fasciola gigantic, fasciola hepatica, GST, RT- PCR

Procedia PDF Downloads 170
333 Materials and Techniques of Anonymous Egyptian Polychrome Cartonnage Mummy Mask: A Multiple Analytical Study

Authors: Hanaa A. Al-Gaoudi, Hassan Ebeid

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The research investigates the materials and processes used in the manufacturing of an Egyptian polychrome cartonnage mummy mask with the aim of dating this object and establishing trade patterns of certain materials that were used and available at the time of ancient Egypt. This anonymous-source object was held in the basement storage of the Egyptian Museum in Cairo (EMC) and has never been on display. Furthermore, there is no information available regarding its owner, provenance, date, and even the time of its possession by the museum. Moreover, the object is in a very poor condition where almost two-thirds of the mask was bent and has never received any previous conservation treatment. This research has utilized well-established multi-analytical methods to identify the considerable diversity of materials that have been used in the manufacturing of this object. These methods include Computed Tomography Scan (CT scan) to acquire detailed pictures of the inside physical structure and condition of the bended layers. Dino-Lite portable digital microscope, scanning electron microscopy with energy dispersive X-ray spectrometer (SEM-EDX), and the non-invasive imaging technique of multispectral imaging (MSI) to obtain information about the physical characteristics and condition of the painted layers and to examine the microstructure of the materials. Portable XRF Spectrometer (PXRF) and X-Ray powder diffraction (XRD) to identify mineral phases and the bulk element composition in the gilded layer, ground, and pigments; Fourier-transform infrared (FTIR) to identify organic compounds and their molecular characterization; accelerator mass spectrometry (AMS 14C) to date the object. Preliminary results suggest that there are no human remains inside the object, and the textile support is linen fibres with tabby weave 1/1 and these fibres are in a very bad condition. Several pigments have been identified, such as Egyptian blue, Magnetite, Egyptian green frit, Hematite, Calcite, and Cinnabar; moreover, the gilded layers are pure gold and the binding media in the pigments is Arabic gum and animal glue in the textile support layer.

Keywords: analytical methods, Egyptian museum, mummy mask, pigments, textile

Procedia PDF Downloads 113
332 An Electrochemical Enzymatic Biosensor Based on Multi-Walled Carbon Nanotubes and Poly (3,4 Ethylenedioxythiophene) Nanocomposites for Organophosphate Detection

Authors: Navpreet Kaur, Himkusha Thakur, Nirmal Prabhakar

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The most controversial issue in crop production is the use of Organophosphate insecticides. This is evident in many reports that Organophosphate (OP) insecticides, among the broad range of pesticides are mainly involved in acute and chronic poisoning cases. OPs detection is of crucial importance for health protection, food and environmental safety. In our study, a nanocomposite of poly (3,4 ethylenedioxythiophene) (PEDOT) and multi-walled carbon nanotubes (MWCNTs) has been deposited electrochemically onto the surface of fluorine doped tin oxide sheets (FTO) for the analysis of malathion OP. The -COOH functionalization of MWCNTs has been done for the covalent binding with amino groups of AChE enzyme. The use of PEDOT-MWCNT films exhibited an excellent conductivity, enables fast transfer kinetics and provided a favourable biocompatible microenvironment for AChE, for the significant malathion OP detection. The prepared biosensors were characterized by Fourier transform infrared spectrometry (FTIR), Field emission-scanning electron microscopy (FE-SEM) and electrochemical studies. Various optimization studies were done for different parameters including pH (7.5), AChE concentration (50 mU), substrate concentration (0.3 mM) and inhibition time (10 min). Substrate kinetics has been performed and studied for the determination of Michaelis Menten constant. The detection limit for malathion OP was calculated to be 1 fM within the linear range 1 fM to 1 µM. The activity of inhibited AChE enzyme was restored to 98% of its original value by 2-pyridine aldoxime methiodide (2-PAM) (5 mM) treatment for 11 min. The oxime 2-PAM is able to remove malathion from the active site of AChE by means of trans-esterification reaction. The storage stability and reusability of the prepared biosensor is observed to be 30 days and seven times, respectively. The application of the developed biosensor has also been evaluated for spiked lettuce sample. Recoveries of malathion from the spiked lettuce sample ranged between 96-98%. The low detection limit obtained by the developed biosensor made them reliable, sensitive and a low cost process.

Keywords: PEDOT-MWCNT, malathion, organophosphates, acetylcholinesterase, biosensor, oxime (2-PAM)

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331 Human TP53 Three Dimentional (3D) Core Domain Hot Spot Mutations at Codon, 36, 72 and 240 are Associated with Oral Squamous Cell Carcinoma

Authors: Saima Saleem, Zubair Abbasi, Abdul Hameed, Mansoor Ahmed Khan, Navid Rashid Qureshi, Abid Azhar

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Oral Squamous Cell Carcinoma (OSCC) is the leading cause of death in the developing countries like Pakistan. This problem aggravates because of the excessive use of available chewing products. In spite of widespread information on their use and purported legislations against their use the Pakistani markets are classical examples of selling chewable carcinogenic mutagens. Reported studies indicated that these products are rich in reactive oxygen species (ROS) and polyphenols. TP53 gene is involved in the suppression of tumor. It has been reported that somatic mutations caused by TP53 gene are the foundation of the cancer. This study aims to find the loss of TP53 functions due to mutation/polymorphism caused by genomic alteration and interaction with tobacco and its related ingredients. Total 260 tissues and blood specimens were collected from OSCC patients and compared with age and sex matched controls. Mutations in exons 2-11 of TP53 were examined by PCR-SSCP. Samples showing mobility shift were directly sequenced. Two mutations were found in exon 4 at nucleotide position 108 and 215 and one in exon 7 at nucleotide position 719 of the coding sequences in patient’s tumor samples. These results show that substitution of proline with arginine at codon 72 and serine with threonine at codon 240 of p53 protein. These polymorphic changes, found in tumor samples of OSCC, could be involved in loss of heterozygocity and apoptotic activity in the binding domain of TP53. The model of the mutated TP53 gene elaborated a nonfunctional unfolded p53 protein, suggesting an important role of these mutations in p53 protein inactivation and malfunction. This nonfunctional 3D model also indicates that exogenous tobacco related carcinogens may act as DNA-damaging agents affecting the structure of DNA. The interpretations could be helpful in establishing the pathways responsible for tumor formation in OSCC patients.

Keywords: TP53 mutation/polymorphism, OSCC, PCR-SSCP, direct DNA sequencing, 3D structure

Procedia PDF Downloads 353
330 Experimental Investigation on Strengthening of Timber Beam Using Glass Fibers and Steel Plates

Authors: Sisaynew Tesfaw Admassu

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The strengthening of timber beams can be necessary for several reasons including the increase of live loads (possible in a historical building for a change of destination of use or upgrading to meet new requirements), the reduction of the resistant cross-sections following deterioration (attacks of biological agents such as fungi, and insects) or traumatic events (fires) and the excess of deflection in the members. The main purpose of strengthening an element is not merely to repair it, but also to prevent and minimize the appearance of future problems. This study did an experimental investigation on the behavior of reference and strengthened solid timber beams. The strengthening materials used in this study were CSM-450 glass fiber and steel materials for both flexural and shear strengthening techniques. Twenty-two solid timber beams of Juniperus procera (TID) species with the dimensions of 60 x 90 x 780 mm were used in the present study. The binding material to bond the strengthening materials with timber was general-purpose resin with Luperox® K10 MEKP catalyst. Three beams were used as control beams (unstrengthen beams) while the remaining nineteen beams were strengthened using the strengthening materials for flexure and shear. All the beams were tested for three points loading to failure by using a Universal Testing Machine, UTM-600kN machine. The experimental results showed that the strengthened beams performed better than the unstrengthen beams. The experimental result of flexural strengthened beams showed that the load-bearing capacity of strengthened beams increased between 16.34 – 42.55%. Four layers of Glass Fiber Reinforced polymer on the tension side of the beams was shown to be the most effective way to enhance load-bearing capacity. The strengthened beams also have an enhancement in their flexural stiffness. The stiffness of flexural strengthened beams was increased between 1.18 – 65.53% as compared to the control beams. The highest increment in stiffness has occurred on beams strengthened using 2x60 mm steel plates. The shear-strengthened beams showed a relatively small amount of performance as compared to flexural-strengthened beams; the reason is that the beams are sufficient for shear. The polyester resin used in the experimental work showed good performance in bonding agents between materials. The resin showed more effectiveness in GFRP materials than steel materials.

Keywords: heritage structures, strengthening, stiffness, adhesive, polyester resin, steel plates

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329 Dielectric Study of Ethanol Water Mixtures at Different Concentration Using Hollow Channel Cantilever Platform

Authors: Maryam S. Ghoraishi, John E. Hawk, Thomas Thundat

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Understanding liquid properties in small scale has become important in recent decades as immerging new microelectromechanical systems (MEMS) devices have been widely used for micro pumps, drug delivery, and many other laboratory-on-microchips analysis. Often in microfluidic devices, fluids are transported electrokinetically. Therefore, extensive knowledge of fluid flow, heat transport, electrokinetics and electrochemistry are key to successful lab on a chip design. Among different microfluidic devices, recently developed hollow channel cantilever offers an ideal platform to study different fluid properties simultaneously without drastic decrease in quality factor which normally occurs when traditional cantilevers operate in the liquid phase. Using hollow channel cantilever, we monitor changes in density and viscosity of liquid while simultaneously investigating dielectric properties of alcohol water binary mixtures. Considerable research has been conducted on alcohol-water mixtures since such a mixture is a typical prototype for biomolecules, Micelle formation, and structural stability of proteins (to name a few). Here we show that hollow channel cantilever can be employed to investigate dielectric properties of ethanol/water mixtures in different concentrations. We study dynamic amplitude shifts of hollow channel cantilever oscillation at different concentrations of ethanol/water for different voltages. Our results show how interactions between solute and solvent, and possibly cluster formation, could change dielectric properties and dipole reorientation of the mixture, as well as the resulting force on the hollow cantilever. For comparison, we also examine higher conductivity ionic mixtures of sodium sulfate solution under the same conditions as low conductivity ethanol/water mixtures. We will show the results from systematic investigation of solvent effects on dielectric properties of the binary mixture. We will also address the question of resolution limits in dielectric study of analyte molecules imposed by solvent concentrations.

Keywords: dielectric constant, cantilever sensors, ethanol water mixtures, low frequency

Procedia PDF Downloads 186
328 Corporate Social Responsibility and the Legal Framework of Foreign Direct Investment: Time for Conceptual Innovation

Authors: Agata Ferreira

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Rapidly increasing debates and initiatives in the area of Corporate Social Responsibility (“CSR”) have reached the world of international investment law. CSR standards that focus on the operations of multinational companies are increasingly relevant in the context of international investment policy making. In the past, the connection between CSR standards and legal framework for foreign direct investment has been largely non-existent. Recently, however, there is a growing trend of a more balance approach to rights and obligations as between investors and states under investment treaties. CSR principles join other social and environmental measures slowly being included in the investment treaties to enhance their sustainable development dimension. Issues of CSR are present on negotiation tables of new mega regional investment treaties like TTIP for example. To date, only a very few bilateral investment treaties and a handful of other international treaties with investment provisions include CSR clauses. In addition, the existing provisions tend to be of a soft type, where parties merely acknowledge importance of good corporate governance and CSR for sustainable development or generally affirm their aim to encourage enterprises to observe internationally recognised guidelines and principles of CSR. The relevant provisions often leave it up to the states to encourage enterprises operating within their territories to voluntarily incorporate CSR principles. The interaction between general non-binding CSR standards, domestic laws and policies and provisions of international investment treaties have not been tested by investment tribunals yet. The role of investment treaties in raising awareness and promoting CSR is still in its infancy. The use of CSR standards in the international investment protection regime for promotion of CSR standards, and as a tool for disciplining investors into complying with such standards, pose a number of questions and is met with resistance from investors` lobbies. Integration of these two areas, CSR and international investment law, both consisting of multilayered, diverse and often overlapping instruments is by no means an easy task. Whether international investment world is ready to embrace CSR standards or shrug them off is a matter of uncertain future. The subject however has been raised, first introductions have been made and the time will show whether the relationship between legal framework of international investment and CSR will flourish or remain dormant.

Keywords: corporate social responsibility, foreign direct investment, investment treaties, sustainable development

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327 Substituted Thiazole Analogues as Anti-Tumor Agents

Authors: Menna Ewida, Dalal Abou El-Ella, Dina Lasheen, Huessin El-Subbagh

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Introduction: Vascular Endothelial Growth Factor receptor (VEGF) is a signal protein produced by cells that stimulates vasculogenesis to create new blood vessels. VEGF family binds to three trans-membrane tyrosine kinase receptors,Dihydrofolate reductase (DHFR) is an enzyme of crucial importance in medicinal chemistry. DHFR catalyzes the reduction 7,8 dihydro-folate to tetrahydrofolate and intimately couples with thymidylate synthase which is a pivotal enzyme that catalysis the reductive methylation of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP) utilizing N5,N10-methylene tetrahydrofolate as a cofactor which functions as the source of the methyl group. Purpose: Novel substituted Thiazole agents were designed as DHFR and VEGF-TK inhibitors with increased synergistic activity and decreased side effects. Methods: Five series of compounds were designed with a rational that mimic the pharmacophoric features present in the reported active compounds that target DHFR & VEGFR. These molecules were docked against Methotrexate & Sorafenib as controls. An in silico ADMET study was also performed to validate the bioavailability of the newly designed compounds. The in silico molecular docking & ADMET study were also applied to the non-classical antifolates for comparison. The interaction energy comparable to that of MTX for DHFRI and Sorafenib for VEGF-TKI activity were recorded. Results: Compound 5 exhibited the highest interaction energy when docked against Sorafenib, While Compound 9 showed the highest interaction energy when docked against MTX with the perfect binding mode. Comparable results were also obtained for the ADMET study. Most of the compounds showed absorption within (95-99) zone which varies according to the type of substituents. Conclusions: The Substituted Thiazole Analogues could be a suitable template for antitumor drugs that possess enhanced bioavailability and act as DHFR and VEGF-TK inhibitors.

Keywords: anti-tumor agents, DHFR, drug design, molecular modeling, VEGFR-TKIs

Procedia PDF Downloads 213