Search results for: bioactive compounds interaction
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 6240

Search results for: bioactive compounds interaction

4440 Detection of Important Biological Elements in Drug-Drug Interaction Occurrence

Authors: Reza Ferdousi, Reza Safdari, Yadollah Omidi

Abstract:

Drug-drug interactions (DDIs) are main cause of the adverse drug reactions and nature of the functional and molecular complexity of drugs behavior in human body make them hard to prevent and treat. With the aid of new technologies derived from mathematical and computational science the DDIs problems can be addressed with minimum cost and efforts. Market basket analysis is known as powerful method to identify co-occurrence of thing to discover patterns and frequency of the elements. In this research, we used market basket analysis to identify important bio-elements in DDIs occurrence. For this, we collected all known DDIs from DrugBank. The obtained data were analyzed by market basket analysis method. We investigated all drug-enzyme, drug-carrier, drug-transporter and drug-target associations. To determine the importance of the extracted bio-elements, extracted rules were evaluated in terms of confidence and support. Market basket analysis of the over 45,000 known DDIs reveals more than 300 important rules that can be used to identify DDIs, CYP 450 family were the most frequent shared bio-elements. We applied extracted rules over 2,000,000 unknown drug pairs that lead to discovery of more than 200,000 potential DDIs. Analysis of the underlying reason behind the DDI phenomena can help to predict and prevent DDI occurrence. Ranking of the extracted rules based on strangeness of them can be a supportive tool to predict the outcome of an unknown DDI.

Keywords: drug-drug interaction, market basket analysis, rule discovery, important bio-elements

Procedia PDF Downloads 296
4439 Effect of Different Contaminants on Mineral Insulating Oil Characteristics

Authors: H. M. Wilhelm, P. O. Fernandes, L. P. Dill, C. Steffens, K. G. Moscon, S. M. Peres, V. Bender, T. Marchesan, J. B. Ferreira Neto

Abstract:

Deterioration of insulating oil is a natural process that occurs during transformers operation. However, this process can be accelerated by some factors, such as oxygen, high temperatures, metals and, moisture, which rapidly reduce oil insulating capacity and favor transformer faults. Parts of building materials of a transformer can be degraded and yield soluble compounds and insoluble particles that shorten the equipment life. Physicochemical tests, dissolved gas analysis (including propane, propylene and, butane), volatile and furanic compounds determination, besides quantitative and morphological analyses of particulate are proposed in this study in order to correlate transformers building materials degradation with insulating oil characteristics. The present investigation involves tests of medium temperature overheating simulation by means of an electric resistance wrapped with the following materials immersed in mineral insulating oil: test I) copper, tin, lead and, paper (heated at 350-400 °C for 8 h); test II) only copper (at 250 °C for 11 h); and test III) only paper (at 250 °C for 8 h and at 350 °C for 8 h). A different experiment is the simulation of electric arc involving copper, using an electric welding machine at two distinct energy sets (low and high). Analysis results showed that dielectric loss was higher in the sample of test I, higher neutralization index and higher values of hydrogen and hydrocarbons, including propane and butane, were also observed. Test III oil presented higher particle count, in addition, ferrographic analysis revealed contamination with fibers and carbonized paper. However, these particles had little influence on the oil physicochemical parameters (dielectric loss and neutralization index) and on the gas production, which was very low. Test II oil showed high levels of methane, ethane, and propylene, indicating the effect of metal on oil degradation. CO2 and CO gases were formed in the highest concentration in test III, as expected. Regarding volatile compounds, in test I acetone, benzene and toluene were detected, which are oil oxidation products. Regarding test III, methanol was identified due to cellulose degradation, as expected. Electric arc simulation test showed the highest oil oxidation in presence of copper and at high temperature, since these samples had huge concentration of hydrogen, ethylene, and acetylene. Particle count was also very high, showing the highest release of copper in such conditions. When comparing high and low energy, the first presented more hydrogen, ethylene, and acetylene. This sample had more similar results to test I, pointing out that the generation of different particles can be the cause for faults such as electric arc. Ferrography showed more evident copper and exfoliation particles than in other samples. Therefore, in this study, by using different combined analytical techniques, it was possible to correlate insulating oil characteristics with possible contaminants, which can lead to transformers failure.

Keywords: Ferrography, gas analysis, insulating mineral oil, particle contamination, transformer failures

Procedia PDF Downloads 206
4438 Theoretical Investigation of the Origin of Interfacial Ferromagnetism of (LaNiO₃)n/(CaMnO₃)m Superlattices

Authors: Jiwuer Jilili, Iogann Tolbatov, Mousumi U. Kahaly

Abstract:

Metal to insulator transition and interfacial magnetism of the LaNiO₃ based superlattice are main interest due to thickness dependent electronic response and tunable magnetic behavior. We investigate the structural, electronic, and magnetic properties of recently experimentally synthesized (LaNiO₃)n/(CaMnO₃)m superlattices with varying LaNiO₃ thickness using density functional theory. The effect of the on-site Coulomb interaction is discussed. In switching from zero to finite U value for Ni atoms, LaNiO₃ shows transitions from half-metallic to metallic character, while spinning ordering changes from paramagnetic to ferromagnetic (FM). For CaMnO₃, U < 3 eV on Mn atoms results in G-type anti-FM spin ordering whereas increasing U value yields FM ordering. In superlattices, metal to insulator transition was achieved with a reduction of LaNiO₃ thickness. The system with one layer of LaNiO₃ yields insulating character. Increasing LaNiO₃ to two layers and above results in the onset of the metallic character with a major contribution from Ni and Mn 3d eg states. Our results for interfacial ferromagnetism, induced Ni magnetic moments and novel antiferromagnetically coupled Ni atoms are consistent with the recent experimental findings. The possible origin of the emergent magnetism is proposed in terms of the exchange interaction and Anderson localization.

Keywords: density functional theory, interfacial magnetism, metal-insulator transition, Ni magnetism.

Procedia PDF Downloads 210
4437 Assessing the Impact of Antiretroviral Mediated Drug-Drug Interactions on Piperaquine Antimalarial Treatment in Pregnant Women Using Physiologically Based Pharmacokinetic Modelling

Authors: Olusola Omolola Olafuyi, Michael Coleman, Raj Kumar Singh Badhan

Abstract:

Introduction: Malaria in pregnancy has morbidity and mortality implication on both mother and unborn child. Piperaquine (PQ) based antimalarial treatment is emerging as a choice antimalarial for pregnant women in the face of resistance to current antimalarial treatment recommendation in pregnancy. Physiological and biochemical changes in pregnant women may affect the pharmacokinetics of the antimalarial drug in these. In malaria endemic regions other infectious diseases like HIV/AIDs are prevalent. Pregnant women who are co-infected with malaria and HIV/AID are at even more greater risk of death not only due to complications of the diseases but also due to drug-drug interactions (DDIs) between antimalarials (AMT) and antiretroviral (ARVs). In this study, physiologically based pharmacokinetic (PBPK) modelling was used to investigate the effect of physiological and biochemical changes on the impact of ARV mediated DDIs in pregnant women in three countries. Method: A PBPK model for PQ was developed on SimCYP® using published physicochemical and pharmacokinetic data of PQ from literature, this was validated in three customized population groups from Thailand, Sudan and Papua New Guinea with clinical data. Validation of PQ model was also done in presence of interaction with efavirenz (pre-validated on SimCYP®). Different albumin levels and pregnancy stages was simulated in the presence of interaction with standard doses of efavirenz and ritonavir. PQ day 7 concentration of 30ng/ml was used as the efficacy endpoint for PQ treatment.. Results: The median day 7 concentration of PQ remained virtually consistent throughout pregnancy and were satisfactory across the three population groups ranging from 26-34.1ng/ml; this implied the efficacy of PQ throughout pregnancy. DDI interaction with ritonavir and efavirenz resulted in modest effect on the day 7 concentrations of PQ with AUCratio ranging from 0.56-0.8 and 1.64-1.79 for efavirenz and ritonavir respectively over 10-40 gestational weeks, however, a reduction in human serum albumin level reflective of severe malaria resulted in significantly reduced the number of subjects attaining the PQ day 7 concentration in the presence of both DDIs. The model demonstrated that the DDI between PQ and ARV in pregnant women with different malaria severities can alter the pharmacokinetic of PQ.

Keywords: antiretroviral, malaria, piperaquine, pregnancy, physiologically-based pharmacokinetics

Procedia PDF Downloads 165
4436 Breast Cancer and BRCA Gene: A Study on Genetic and Environmental Interaction

Authors: Abhishikta Ghosh Roy

Abstract:

Breast cancer is the most common malignancy among women globally, including India. Human breast cancer results from the genetic and environmental interaction. The present study attempts to understand the molecular heterogeneity of BRCA1 and BRCA2 genes, as well as to understand the association of various lifestyle and reproductive variables for the Breast Cancer risk. The study was conducted amongst 110 patients and 128 controls with total DNA sequencing of flanking and coding regions of BRCA1 BRCA2 genes that revealed ten Single Nucleotide Polymorphisms (SNPs) (6 novels). The controls selected for the study were age, sex and ethnic group matched. After written and informed consent biological samples were collected from the subjects. After detailed molecular analysis, significant (p < 0.005) molecular heterogeneity is revealed in terms of SNPs in BRCA1 (4 Exonic & 1 Intronic) and BRCA2 (2exonic and 3 Intronic) genes. The augmentation study investigated significant (p < 0.05) association with positive family history, early age at menarche, irregular menstrual periods, menopause, prolong contraceptive use, nulliparity, history of abortions, consumption of alcohol and smoking for breast cancer risk. To the best of authors knowledge, this study is the first of its kind, envisaged that the identification of the SNPs and modification of the lifestyle factors might aid to minimize the risk among the Bengalee Hindu females.

Keywords: breast cancer, BRCA, lifestyle, India

Procedia PDF Downloads 103
4435 Efficient Synthesis of Thiourea Based Iminothiazoline Heterocycles

Authors: Hummera Rafique, Aamer Saeed

Abstract:

Thioureas are highly biologically active compounds, as many important applications are associated with this nucleus. They serve as exceptionally versatile building block for the synthesis of wide variety of heterocyclic systems, which also possess extensive range of bioactivities. These thioureas were converted into five-membered heterocycles with imino moiety like ethyl 4-[2-benzamido-4-methylthiazol-3(2H)-yl)]benzoates (2a-j) by base catalyzed cyclization of corresponding thioureas with 2-bromoacetone and triethylamine in good yields.

Keywords: ethyl 4-[2-benzamido-4-methylthiazol-3(2H)-yl)]benzoates, ethyl 4-(3-benzoylthioureido) benzoates, antibacterial activity

Procedia PDF Downloads 343
4434 Drying Kinetics, Energy Requirement, Bioactive Composition, and Mathematical Modeling of Allium Cepa Slices

Authors: Felix U. Asoiro, Meshack I. Simeon, Chinenye E. Azuka, Harami Solomon, Chukwuemeka J. Ohagwu

Abstract:

The drying kinetics, specific energy consumed (SEC), effective moisture diffusivity (EMD), flavonoid, phenolic, and vitamin C contents of onion slices dried under convective oven drying (COD) were compared with microwave drying (MD). Drying was performed with onion slice thicknesses of 2, 4, 6, and 8 mm; air drying temperatures of 60, 80, and 100°C for COD, and microwave power of 450 W for MD. A decrease in slice thickness and an increase in drying air temperature led to a drop in the drying time. As thickness increased from 2 – 8 mm, EMD rose from 1.1-4.35 x 10⁻⁸ at 60°C, 1.1-5.6 x 10⁻⁸ at 80°C, and 1.25-6.12 x 10⁻⁸ at 100°C with MD treatments yielding the highest mean value (6.65 x 10⁻⁸ m² s⁻¹) at 8 mm. Maximum SEC for onion slices in COD was 238.27 kWh/kg H₂O (2 mm thickness), and the minimum was 39.4 kWh/kg H₂O (8 mm thickness) whereas maximum during MD was 25.33 kWh/kg H₂O (8 mm thickness) and minimum, 18.7 kWh/kg H₂O (2 mm thickness). MD treatment gave a significant (p 0.05) increase in the flavonoid (39.42 – 64.4%), phenolic (38.0 – 46.84%), and vitamin C (3.7 – 4.23 mg 100 g⁻¹) contents, while COD treatment at 60°C and 100°C had positive effects on only vitamin C and phenolic contents, respectively. In comparison, the Weibull model gave the overall best fit (highest R²=0.999; lowest SSE=0.0002, RSME=0.0123, and χ²= 0.0004) when drying 2 mm onion slices at 100°C.

Keywords: allium cepa, drying kinetics, specific energy consumption, flavonoid, vitamin C, microwave oven drying

Procedia PDF Downloads 118
4433 Preparation and Characterization of AlkylAmines’ Surface Functionalized Activated Carbons for Dye Removal

Authors: Said M. AL-Mashaikhi, El-Said I. El-Shafey, Fakhreldin O. Suliman, Saleh Al-Busafi

Abstract:

Activated carbon (AC) was prepared from date palm leaflets via NaOH activation. AC was oxidized using nitric acid, producing oxidized activated carbon (OAC). OAC was surface functionalized using different amine surfactants, including methylamine (ONM), ethylamine (ONE), and diethylamine (ONDE) using the amide coupling process. Produced carbons were surface characterized for surface area and porosity, X-ray diffraction, SEM, FTIR, and TGA. AC surface area (580 m²/g) has shown a decrease in oxidation to 260 m²/g for OAC. On amine functionalization, the surface area has further decreased to 218, 108, and 20 m²/g on functionalization with methylamine, ethylamine, and diethylamine, respectively. FTIR and TGA showed that the nature of amine functionalization of AC is chemical. Methylene blue sorption was tested on these carbons in terms of kinetics and equilibrium. Sorption was found faster on amine-functionalized carbons than both AC and OAC, and this is due to hydrophobic interaction with the alkyl groups immobilized with data following pseudo second-order reaction. On the other hand, AC showed the slowest adsorption kinetic process due to the diffusion in the porous structure of AC. Sorption equilibrium data was found to follow the Langmuir sorption isotherm with maximum sorption found on ONE. Regardless of its lower surface area than activated carbon, ethylamine functionalized AC showed better performance than AC in terms of kinetics and equilibrium for dye removal.

Keywords: activated carbon, dye removal, functionalization, hydrophobic interaction, water treatment

Procedia PDF Downloads 147
4432 Molecular Topology and TLC Retention Behaviour of s-Triazines: QSRR Study

Authors: Lidija R. Jevrić, Sanja O. Podunavac-Kuzmanović, Strahinja Z. Kovačević

Abstract:

Quantitative structure-retention relationship (QSRR) analysis was used to predict the chromatographic behavior of s-triazine derivatives by using theoretical descriptors computed from the chemical structure. Fundamental basis of the reported investigation is to relate molecular topological descriptors with chromatographic behavior of s-triazine derivatives obtained by reversed-phase (RP) thin layer chromatography (TLC) on silica gel impregnated with paraffin oil and applied ethanol-water (φ = 0.5-0.8; v/v). Retention parameter (RM0) of 14 investigated s-triazine derivatives was used as dependent variable while simple connectivity index different orders were used as independent variables. The best QSRR model for predicting RM0 value was obtained with simple third order connectivity index (3χ) in the second-degree polynomial equation. Numerical values of the correlation coefficient (r=0.915), Fisher's value (F=28.34) and root mean square error (RMSE = 0.36) indicate that model is statistically significant. In order to test the predictive power of the QSRR model leave-one-out cross-validation technique has been applied. The parameters of the internal cross-validation analysis (r2CV=0.79, r2adj=0.81, PRESS=1.89) reflect the high predictive ability of the generated model and it confirms that can be used to predict RM0 value. Multivariate classification technique, hierarchical cluster analysis (HCA), has been applied in order to group molecules according to their molecular connectivity indices. HCA is a descriptive statistical method and it is the most frequently used for important area of data processing such is classification. The HCA performed on simple molecular connectivity indices obtained from the 2D structure of investigated s-triazine compounds resulted in two main clusters in which compounds molecules were grouped according to the number of atoms in the molecule. This is in agreement with the fact that these descriptors were calculated on the basis of the number of atoms in the molecule of the investigated s-triazine derivatives.

Keywords: s-triazines, QSRR, chemometrics, chromatography, molecular descriptors

Procedia PDF Downloads 376
4431 Information Communication Technologies and Renewable Technologies' Impact on Irish People's Lifestyle: A Constructivist Grounded Theory Study

Authors: Hamilton V. Niculescu

Abstract:

This paper discusses findings relating to people's engagement with mobile communication technologies and remote automated systems. This interdisciplinary study employs a constructivist grounded theory methodology, with qualitative data that was generated following in-depth semi-structured interviews with 18 people living in Ireland being corroborated with participants' observations and quantitative data. Additional data was collected following participants' remote interaction with six custom-built automated enclosures, located at six different sites around Dublin, Republic of Ireland. This paper argues that ownership and education play a vital role in people engaging with and adoption of new technologies. Analysis of participants' behavior and attitude towards Information Communication Technologies (ICT) suggests that innovations do not always improve peoples' social inclusion. Technological innovations are sometimes perceived as destroying communities and create a dysfunctional society. Moreover, the findings indicate that a lack of public information and support from Irish governmental institutions, as well as limited off-the-shelves availability, has led to low trust and adoption of renewable technologies. A limited variation in participants' behavior and interaction patterns with technologies was observed during the study. This suggests that people will eventually adopt new technologies according to their needs and experience, even though they initially rejected the idea of changing their lifestyle.

Keywords: automation, communication, ICT, renewables

Procedia PDF Downloads 94
4430 Phytoremediation-A Plant Based Cleansing Method to Obtain Quality Medicinal Plants and Natural Products

Authors: Hannah S. Elizabeth, D. Gnanasekaran, M. R. Manju Gowda, Antony George

Abstract:

Phytoremediation a new technology of remediating the contaminated soil, water and air using plants and serves as a green technology with environmental friendly approach. The main aim of this technique is cleansing and detoxifying of organic compounds, organo-phosphorous pesticides, heavy metals like arsenic, iron, cadmium, gold, radioactive elements which cause teratogenic and life threatening diseases to mankind and animal kingdom when consume the food crops, vegetables, fruits, cerals, and millets obtained from the contaminated soil. Also, directly they may damage the genetic materials thereby alters the biosynthetic pathways of secondary metabolites and other phytoconstituents which may have different pharmacological activities which lead to lost their efficacy and potency as well. It would reflect in mutagenicity, drug resistance and affect other antagonistic properties of normal metabolism. Is the technology for real clean-up of contaminated soils and the contaminants which are potentially toxic. It reduces the risks produced by a contaminated soil by decreasing contaminants using plants as a source. The advantages are cost-effectiveness and less ecosystem disruption. Plants may also help to stabilize contaminants by accumulating and precipitating toxic trace elements in the roots. Organic pollutants can potentially be chemically degraded and ultimately mineralized into harmless biological compounds. Hence, the use of plants to revitalize contaminated sites is gaining more attention and preferred for its cost-effective when compared to other chemical methods. The introduction of harmful substances into the environment has been shown to have many adverse effects on human health, agricultural productivity, and natural ecosystems. Because the costs of growing a crop are minimal compared to those of soil removal and replacement, the use of plants to remediate hazardous soils is seen as having great promise.

Keywords: cost effective, eco-friendly, phytoremediation, secondary metabolites

Procedia PDF Downloads 265
4429 A Brief of Survey on Use of Videoconferencing in Teaching during Quarantine Conducted in Sao Paulo

Authors: Fernanda Laureti T. Ferreira, Kazuo Nishimoto

Abstract:

This paper presents a summary of the experience on videoconferencing tools that have been used to teach regular classes during this pandemic period in educational institutions in São Paulo, which tools and applications are most used and the challenges related to this mode of delivery. At this moment, the massive online education is not a choice of students or a structured development of education system, but a solution that emerged to attend urgent needs and it presents the opportunity to teach and learning available for the most students in this single time of social isolation that forced among others, this significant change for education, students, teachers, institutions and families. Distance education enables synchronous and asynchronous mode classes, and even though the current circumstances generate discomfort and uncertainty, on the other hand, there is a chance to promote a 'learning to learn'. The videoconference is a preferred choice of schools because synchronous mode to give more interaction between a group of students and teachers, but this mode requires specifics teacher competencies and skills, in addition to equipment and provision of adequate internet signal for all participants of the process. The approach is making use of known technical information about video conference tools and the results of search answered by a group of students, teachers, schools, and parents. The results presented refer to the perspectives of students and parents as respondents.

Keywords: distance education, interaction on education, online classes, synchronous e-learning, videoconference

Procedia PDF Downloads 111
4428 Emergentist Metaphorical Creativity: Towards a Model of Analysing Metaphorical Creativity in Interactive Talk

Authors: Afef Badri

Abstract:

Metaphorical creativity does not constitute a static property of discourse. It is an interactive dynamic process created online. There has been a lack of research concerning online produced metaphorical creativity. This paper intends to account for metaphorical creativity in online talk-in-interaction as a dynamic process that emerges as discourse unfolds. It brings together insights from the emergentist approach to the study of metaphor in verbal interactions and insights from conceptual blending approach as a model for analysing online metaphorical constructions to propose a model for studying metaphorical creativity in interactive talk. The model is based on three focal points. First, metaphorical creativity is a dynamic emergent and open-to-change process that evolves in real time as interlocutors constantly blend and re-blend previous metaphorical contributions. Second, it is not a product of isolated individual minds but a joint achievement that is co-constructed and co-elaborated by interlocutors. The third and most important point is that the emergent process of metaphorical creativity is tightly shaped by contextual variables surrounding talk-in-interaction. It is grounded in the framework of interpretation of interlocutors. It is constrained by preceding contributions in a way that creates textual cohesion of the verbal exchange and it is also a goal-oriented process predefined by the communicative intention of each participant in a way that reveals the ideological coherence/incoherence of the entire conversation.

Keywords: communicative intention, conceptual blending, the emergentist approach, metaphorical creativity

Procedia PDF Downloads 247
4427 Designed Purine Molecules and in-silico Evaluation of Aurora Kinase Inhibition in Breast Cancer

Authors: Pooja Kumari, Anandkumar Tengli

Abstract:

Aurora kinase enzyme, a protein on overexpression, leads to metastasis and is extremely important for women’s health in terms of prevention or treatment. While creating a targeted technique, the aim of the work is to design purine molecules that inhibit in aurora kinase enzyme and helps to suppress breast cancer. Purine molecules attached to an amino acid in DNA block protein synthesis or halt the replication and metastasis caused by the aurora kinase enzyme. Various protein related to the overexpression of aurora protein was docked with purine molecule using Biovia Drug Discovery, the perpetual software. Various parameters like X-ray crystallographic structure, presence of ligand, Ramachandran plot, resolution, etc., were taken into consideration for selecting the target protein. A higher negative binding scored molecule has been taken for simulation studies. According to the available research and computational analyses, purine compounds may be powerful enough to demonstrate a greater affinity for the aurora target. Despite being clinically effective now, purines were originally meant to fight breast cancer by inhibiting the aurora kinase enzyme. In in-silico studies, it is observed that purine compounds have a moderate to high potency compared to other molecules, and our research into the literature revealed that purine molecules have a lower risk of side effects. The research involves the design, synthesis, and identification of active purine molecules against breast cancer. Purines are structurally similar to the normal metabolites of adenine and guanine; hence interfere/compete with protein synthesis and suppress the abnormal proliferation of cells/tissues. As a result, purine target metastasis cells and stop the growth of kinase; purine derivatives bind with DNA and aurora protein which may stop the growth of protein or inhibits replication and stop metastasis of overexpressed aurora kinase enzyme.

Keywords: aurora kinases, in silico studies, medicinal chemistry, combination therapies, chronic cancer, clinical translation

Procedia PDF Downloads 71
4426 The Regulation of the Cancer Epigenetic Landscape Lies in the Realm of the Long Non-coding RNAs

Authors: Ricardo Alberto Chiong Zevallos, Eduardo Moraes Rego Reis

Abstract:

Pancreatic adenocarcinoma (PDAC) patients have a less than 10% 5-year survival rate. PDAC has no defined diagnostic and prognostic biomarkers. Gemcitabine is the first-line drug in PDAC and several other cancers. Long non-coding RNAs (lncRNAs) contribute to the tumorigenesis and are potential biomarkers for PDAC. Although lncRNAs aren’t translated into proteins, they have important functions. LncRNAs can decoy or recruit proteins from the epigenetic machinery, act as microRNA sponges, participate in protein translocation through different cellular compartments, and even promote chemoresistance. The chromatin remodeling enzyme EZH2 is a histone methyltransferase that catalyzes the methylation of histone 3 at lysine 27, silencing local expression. EZH2 is ambivalent, it can also activate gene expression independently of its histone methyltransferase activity. EZH2 is overexpressed in several cancers and interacts with lncRNAs, being recruited to a specific locus. EZH2 can be recruited to activate an oncogene or silence a tumor suppressor. The lncRNAs misregulation in cancer can result in the differential recruitment of EZH2 and in a distinct epigenetic landscape, promoting chemoresistance. The relevance of the EZH2-lncRNAs interaction to chemoresistant PDAC was assessed by Real Time quantitative PCR (RT-qPCR) and RNA Immunoprecipitation (RIP) experiments with naïve and gemcitabine-resistant PDAC cells. The expression of several lncRNAs and EZH2 gene targets was evaluated contrasting naïve and resistant cells. Selection of candidate genes was made by bioinformatic analysis and literature curation. Indeed, the resistant cell line showed higher expression of chemoresistant-associated lncRNAs and protein coding genes. RIP detected lncRNAs interacting with EZH2 with varying intensity levels in the cell lines. During RIP, the nuclear fraction of the cells was incubated with an antibody for EZH2 and with magnetic beads. The RNA precipitated with the beads-antibody-EZH2 complex was isolated and reverse transcribed. The presence of candidate lncRNAs was detected by RT-qPCR, and the enrichment was calculated relative to INPUT (total lysate control sample collected before RIP). The enrichment levels varied across the several lncRNAs and cell lines. The EZH2-lncRNA interaction might be responsible for the regulation of chemoresistance-associated genes in multiple cancers. The relevance of the lncRNA-EZH2 interaction to PDAC was assessed by siRNA knockdown of a lncRNA, followed by the analysis of the EZH2 target expression by RT-qPCR. The chromatin immunoprecipitation (ChIP) of EZH2 and H3K27me3 followed by RT-qPCR with primers for EZH2 targets also assess the specificity of the EZH2 recruitment by the lncRNA. This is the first report of the interaction of EZH2 and lncRNAs HOTTIP and PVT1 in chemoresistant PDAC. HOTTIP and PVT1 were described as promoting chemoresistance in several cancers, but the role of EZH2 is not clarified. For the first time, the lncRNA LINC01133 was detected in a chemoresistant cancer. The interaction of EZH2 with LINC02577, LINC00920, LINC00941, and LINC01559 have never been reported in any context. The novel lncRNAs-EZH2 interactions regulate chemoresistant-associated genes in PDAC and might be relevant to other cancers. Therapies targeting EZH2 alone weren’t successful, and a combinatorial approach also targeting the lncRNAs interacting with it might be key to overcome chemoresistance in several cancers.

Keywords: epigenetics, chemoresistance, long non-coding RNAs, pancreatic cancer, histone modification

Procedia PDF Downloads 74
4425 Development of a Real-Time Simulink Based Robotic System to Study Force Feedback Mechanism during Instrument-Object Interaction

Authors: Jaydip M. Desai, Antonio Valdevit, Arthur Ritter

Abstract:

Robotic surgery is used to enhance minimally invasive surgical procedure. It provides greater degree of freedom for surgical tools but lacks of haptic feedback system to provide sense of touch to the surgeon. Surgical robots work on master-slave operation, where user is a master and robotic arms are the slaves. Current, surgical robots provide precise control of the surgical tools, but heavily rely on visual feedback, which sometimes cause damage to the inner organs. The goal of this research was to design and develop a real-time simulink based robotic system to study force feedback mechanism during instrument-object interaction. Setup includes three Velmex XSlide assembly (XYZ Stage) for three dimensional movement, an end effector assembly for forceps, electronic circuit for four strain gages, two Novint Falcon 3D gaming controllers, microcontroller board with linear actuators, MATLAB and Simulink toolboxes. Strain gages were calibrated using Imada Digital Force Gauge device and tested with a hard-core wire to measure instrument-object interaction in the range of 0-35N. Designed simulink model successfully acquires 3D coordinates from two Novint Falcon controllers and transfer coordinates to the XYZ stage and forceps. Simulink model also reads strain gages signal through 10-bit analog to digital converter resolution of a microcontroller assembly in real time, converts voltage into force and feedback the output signals to the Novint Falcon controller for force feedback mechanism. Experimental setup allows user to change forward kinematics algorithms to achieve the best-desired movement of the XYZ stage and forceps. This project combines haptic technology with surgical robot to provide sense of touch to the user controlling forceps through machine-computer interface.

Keywords: surgical robot, haptic feedback, MATLAB, strain gage, simulink

Procedia PDF Downloads 520
4424 Automatic Fluid-Structure Interaction Modeling and Analysis of Butterfly Valve Using Python Script

Authors: N. Guru Prasath, Sangjin Ma, Chang-Wan Kim

Abstract:

A butterfly valve is a quarter turn valve which is used to control the flow of a fluid through a section of pipe. Generally, butterfly valve is used in wide range of applications such as water distribution, sewage, oil and gas plants. In particular, butterfly valve with larger diameter finds its immense applications in hydro power plants to control the fluid flow. In-lieu with the constraints in cost and size to run laboratory setup, analysis of large diameter values will be mostly studied by computational method which is the best and inexpensive solution. For fluid and structural analysis, CFD and FEM software is used to perform large scale valve analyses, respectively. In order to perform above analysis in butterfly valve, the CAD model has to recreate and perform mesh in conventional software’s for various dimensions of valve. Therefore, its limitation is time consuming process. In-order to overcome that issue, python code was created to outcome complete pre-processing setup automatically in Salome software. Applying dimensions of the model clearly in the python code makes the running time comparatively lower and easier way to perform analysis of the valve. Hence, in this paper, an attempt was made to study the fluid-structure interaction (FSI) of butterfly valves by varying the valve angles and dimensions using python code in pre-processing software, and results are produced.

Keywords: butterfly valve, flow coefficient, automatic CFD analysis, FSI analysis

Procedia PDF Downloads 227
4423 From Wave-Powered Propulsion to Flight with Membrane Wings: Insights Powered by High-Fidelity Immersed Boundary Methods based FSI Simulations

Authors: Rajat Mittal, Jung Hee Seo, Jacob Turner, Harshal Raut

Abstract:

The perpetual advancement in computational capabilities, coupled with the continuous evolution of software tools and numerical algorithms, is creating novel avenues for research, exploration, and application at the nexus of computational fluid and structural mechanics. Fish leverage their remarkably flexible bodies and fins to harness energy from vortices, propelling themselves with an elegance and efficiency that captivates engineers. Bats fly with unparalleled agility and speed by using their flexible membrane wings. Wave-assisted propulsion (WAP) systems, utilizing elastically mounted hydrofoils, convert wave energy into thrust. Each of these problems involves a complex and elegant interplay between fluid dynamics and structural mechanics. Historically, investigations into such phenomena were constrained by available tools, but modern computational advancements now facilitate exploration of these multi-physics challenges with an unprecedented level of fidelity, precision, and realism. In this work, the author will discuss projects that harness the capabilities of high-fidelity sharp-interface immersed boundary methods to address a spectrum of engineering and biological challenges involving fluid-structure interaction.

Keywords: immersed boundary methods, CFD, bioflight, fluid structure interaction

Procedia PDF Downloads 47
4422 Conformation Prediction of Human Plasmin and Docking on Gold Nanoparticle

Authors: Wen-Shyong Tzou, Chih-Ching Huang, Chin-Hwa Hu, Ying-Tsang Lo, Tun-Wen Pai, Chia-Yin Chiang, Chung-Hao Li, Hong-Jyuan Jian

Abstract:

Plasmin plays an important role in the human circulatory system owing to its catalytic ability of fibrinolysis. The immediate injection of plasmin in patients of strokes has intrigued many scientists to design vectors that can transport plasmin to the desired location in human body. Here we predict the structure of human plasmin and investigate the interaction of plasmin with the gold-nanoparticle. Because the crystal structure of plasminogen has been solved, we deleted N-terminal domain (Pan-apple domain) of plasminogen and generate a mimic of the active form of this enzyme (plasmin). We conducted a simulated annealing process on plasmin and discovered a very large conformation occurs. Kringle domains 1, 4 and 5 had been observed to leave its original location relative to the main body of the enzyme and the original doughnut shape of this enzyme has been transformed to a V-shaped by opening its two arms. This observation of conformational change is consistent with the experimental results of neutron scattering and centrifugation. We subsequently docked the plasmin on the simulated gold surface to predict their interaction. The V-shaped plasmin could utilize its Kringle domain and catalytic domain to contact the gold surface. Our findings not only reveal the flexibility of plasmin structure but also provide a guide for the design of a plasmin-gold nanoparticle.

Keywords: docking, gold nanoparticle, molecular simulation, plasmin

Procedia PDF Downloads 462
4421 [Keynote Speech]: Risk Management during the Rendition Process: Use of Screen-Voice Recordings in Translator Training

Authors: Maggie Hui

Abstract:

Risk management is not a new concept; however, it is an uncharted area as applied to the translation process and translator training. Serving as one of the self-discovery activities in their practicum course, a two-cycle experiment was carried out with a class of 13 MA translation students with an attempt to explore their risk management while translating in a simulated setting that involves translator-client relations. To test the effects of the main variable of translators’ interaction with the simulated clients, the researcher employed control-group translators and two experiment groups (with Group A being the translator in Cycle 1 and the client in Cycle 2, and Group B on the client position in Cycle 1 and the translator position in Cycle 2). Experiment cycle 1 aims to explore if there would be any behavioral difference in risk management between translators with interaction with the simulated clients, i.e. experiment group A, and their counterparts without such interaction, i.e. control group. Design of Cycle 2 concerns the order of playing different roles of the translator and client in the experiment, and provides information to compare behavior of translators of the two experiment groups. Since this is process-oriented research, it is necessary to hypothesize what was happening in the translators’ minds. The researcher made use of a user-friendly screen-voice recording freeware to record subjects’ screen activities, including every word the translator typed and every change they made to the rendition, the websites they browsed and the reference tools they used, in addition to the verbalization of their thoughts throughout the process. The research observes the translation procedures subjects considered and finally adopted, and looks into the justifications for their procedures, in order to interpret their risk management. The qualitative and quantitative results of this study have some implications for translator training: (a) the experience of being a client seems to reinforce the translator’s risk aversion; (b) the use of role-playing simulation can empower students’ learning by enhancing their attitudinal or psycho-physiological competence, interpersonal competence and strategic competence; and (c) the screen-voice recordings serve as a helpful tool for learners to reflect on their rendition processes, i.e. what they performed satisfactorily and unsatisfactorily while translating and what they could do for improvement in future translation tasks.

Keywords: risk management, screen-voice recordings, simulated translator-client relations, translation pedagogy, translation process-oriented research

Procedia PDF Downloads 252
4420 Computational Approach for Grp78–Nf-ΚB Binding Interactions in the Context of Neuroprotective Pathway in Brain Injuries

Authors: Janneth Gonzalez, Marco Avila, George Barreto

Abstract:

GRP78 participates in multiple functions in the cell during normal and pathological conditions, controlling calcium homeostasis, protein folding and unfolded protein response. GRP78 is located in the endoplasmic reticulum, but it can change its location under stress, hypoxic and apoptotic conditions. NF-κB represents the keystone of the inflammatory process and regulates the transcription of several genes related with apoptosis, differentiation, and cell growth. The possible relationship between GRP78-NF-κB could support and explain several mechanisms that may regulate a variety of cell functions, especially following brain injuries. Although several reports show interactions between NF-κB and heat shock proteins family members, there is a lack of information on how GRP78 may be interacting with NF-κB, and possibly regulating its downstream activation. Therefore, we assessed the computational predictions of the GRP78 (Chain A) and NF-κB complex (IkB alpha and p65) protein-protein interactions. The interaction interface of the docking model showed that the amino acids ASN 47, GLU 215, GLY 403 of GRP78 and THR 54, ASN 182 and HIS 184 of NF-κB are key residues involved in the docking. The electrostatic field between GRP78-NF-κB interfaces and molecular dynamic simulations support the possible interaction between the proteins. In conclusion, this work shed some light in the possible GRP78-NF-κB complex indicating key residues in this crosstalk, which may be used as an input for better drug design strategy targeting NF-κB downstream signaling as a new therapeutic approach following brain injuries.

Keywords: computational biology, protein interactions, Grp78, bioinformatics, molecular dynamics

Procedia PDF Downloads 330
4419 High Level Expression of Fluorinase in Escherichia Coli and Pichia Pastoris

Authors: Lee A. Browne, K. Rumbold

Abstract:

The first fluorinating enzyme, 5'-fluoro-5'-deoxyadenosine synthase (fluorinase) was isolated from the soil bacterium Streptomyces cattleya. Such an enzyme, with the ability to catalyze a C-F bond, presents great potential as a biocatalyst. Naturally fluorinated compounds are extremely rare in nature. As a result, the number of fluorinases identified remains relatively few. The field of fluorination is almost completely synthetic. However, with the increasing demand for fluorinated organic compounds of commercial value in the agrochemical, pharmaceutical and materials industries, it has become necessary to utilize biologically based methods such as biocatalysts. A key step in this crucial process is the large-scale production of the fluorinase enzyme in considerable quantities for industrial applications. Thus, this study aimed to optimize expression of the fluorinase enzyme in both prokaryotic and eukaryotic expression systems in order to obtain high protein yields. The fluorinase gene was cloned into the pET 41b(+) and pPinkα-HC vectors and used to transform the expression hosts, E.coli BL21(DE3) and Pichia pastoris (PichiaPink™ strains) respectively. Expression trials were conducted to select optimal conditions for expression in both expression systems. Fluorinase catalyses a reaction between S-adenosyl-L-Methionine (SAM) and fluoride ion to produce 5'-fluorodeoxyadenosine (5'FDA) and L-Methionine. The activity of the enzyme was determined using HPLC by measuring the product of the reaction 5'FDA. A gradient mobile phase of 95:5 v/v 50mM potassium phosphate buffer to a final mobile phase containing 80:20 v/v 50mM potassium phosphate buffer and acetonitrile were used. This resulted in the complete separation of SAM and 5’-FDA which eluted at 1.3 minutes and 3.4 minutes respectively. This proved that the fluorinase enzyme was active. Optimising expression of the fluorinase enzyme was successful in both E.coli and PichiaPink™ where high expression levels in both expression systems were achieved. Protein production will be scaled up in PichiaPink™ using fermentation to achieve large-scale protein production. High level expression of protein is essential in biocatalysis for the availability of enzymes for industrial applications.

Keywords: biocatalyst, expression, fluorinase, PichiaPink™

Procedia PDF Downloads 539
4418 The Impact of Garlic and Citrus Extracts on Energy Retention and Methane Production in Ruminants in vitro

Authors: Michael Graz, Natasha Hurril, Andrew Shearer

Abstract:

Research on feed supplementation with natural compounds is currently being intensively pursued with a view to improving energy utilisation in ruminants and mitigating the production of methane by these animals. Towards this end, a novel combination of extracts from garlic and bitter orange was therefore selected for trials on the basis of their previously published in vitro anti-methanogenic potential. Three separate in vitro experiments were conducted to determine energy utilisation and greenhouse gas production. These included use of rumen fluid from fistulated cows and sheep in batch culture, the Hohenheim gas test, and the Rusitec technique. Experimental and control arms were utilised, with 5g extracts per kilogram of total dietary dry matter (0.05g/kg active compounds) being used to supplement or not supplement the in vitro systems. Respiratory measurements were conducted on experimental day 1 for the batch culture and Hohenheim gas test and on day 14-21 for the Rusitec Technique (in a 21-day trial). Measurements included methane (CH4) production, total volatile fatty acid (VFA) concentration, molar proportions of acetate, propionate and butyrate and degradation of organic matter (Rusitec). CH4 production was reduced by 82% (±16%), 68% (±11%) and 37% (±4%) in the batch culture, Hohenheim gas test and Rusitec, respectively. Total VFA production was reduced by 13% (±2%) and 2% (±0.1%) in the batch culture and Hohenheim gas test whilst it was increased by 8% (±2%) in the Rusitec. Total VFA production was reduced in all tests between 2 and 10%, whilst acetate production was reduced between 10% and 29%. Propionate production which is an indicator of weight gain was increased in all cases between 16% and 30%. Butyrate production which is considered an indicator of potential milk yield was increased by between 6 and 11%. Degradation of organic matter in the Rusitec experiments was improved by 10% (±0.1%). In conclusion, the study demonstrated the potential of the combination of garlic and citrus extracts to improve digestion, enhance body energy retention and limit CH4 formation in relation to feed intake.

Keywords: citrus, garlic, methane, ruminants

Procedia PDF Downloads 319
4417 Comparative Study of Antimicrobial, Antioxidant and Physicochemical Properties of Four Culinary Herbs Grown in Sri Lanka

Authors: Thilini Kananke

Abstract:

Culinary herbs have long been considered as significant dietary sources of many potential health-promoting compounds. The present research focused on analysis of antimicrobial, antioxidant and physicochemical properties in selected four culinary herbs namely Murraya koenigii (Curry leaves), Pandanus amaryllifolius (Pandan leaves), Cymbopogon citrates (Lemon grass leaves), and Mentha Piperita (Minchi leaves) obtained from several market sites in Ratnapura District, Sri Lanka. The antimicrobial activity of ethanolic, chloroform and distilled water extracts of culinary herbs were evaluated against the strains of Staphylococcus aureus, Salmonella typhi and Shigella spp. Total phenolic content and the radical scavenging activity (using DPPH assay) of culinary herbs were determined. Four heavy metals (Cu, Cd, Pb and Fe) were analyzed in the selected culinary herbs using the atomic absorption spectroscopy (AAS). Proximate compositions of the selected herbs were analyzed using AOAC official methods. Antimicrobial activity of all selected culinary herbs showed relativity high inhibition zones against S. aureus. Pandan leaves showed the least antimicrobial activity against selected bacterial strains compared with other culinary herbs. Both the highest radical scavenging activity (lower IC50 value) and the total phenolic content (25.57 ±3.54µg GAE/100g) were reported in Mentha piperita extract. The highest concentrations of Cu, Fe and Cd were reported in Curry leaves (29.15 mg/kg), Lemon grass leaves (257.98 mg/kg) and Pandan leaves (6.05 mg/kg) respectively. The heavy metal contents detected in all culinary herbs were below the permitted limits set by WHO/FAO, except Cd. The highest moisture (85.00±0.00%) and fiber (10.66± 2.00%) contents were found in Pandan leaves, while the highest protein (8.94±0.29%), fat (12.3± 2.52%) and ash (3.50± 0.17%) contents were reported in curry leaves. The information obtained from this study highlights the importance of further investigation of other antioxidant, antimicrobial and health promoting compounds of culinary herbs available in Sri Lanka for a detailed comparison.

Keywords: antimicrobial, antioxidant, culinary herbs, proximate analysis

Procedia PDF Downloads 157
4416 Seismic Integrity Determination of Dams in Urban Areas

Authors: J. M. Mayoral, M. Anaya

Abstract:

The urban and economic development of cities demands the construction of water use and flood control infrastructure. Likewise, it is necessary to determine the safety level of the structures built with the current standards and if it is necessary to define the reinforcement actions. The foregoing is even more important in structures of great importance, such as dams, since they imply a greater risk for the population in case of failure or undesirable operating conditions (e.g., seepage, cracks, subsidence). This article presents a methodology for determining the seismic integrity of dams in urban areas. From direct measurements of the dynamic properties using geophysical exploration and ambient seismic noise measurements, the seismic integrity of the concrete-faced rockfill dam selected as a case of study is evaluated. To validate the results, two accelerometer stations were installed (e.g., free field and crest of the dam). Once the dynamic properties were determined, three-dimensional finite difference models were developed to evaluate the dam seismic performance for different intensities of movement, considering the site response and soil-structure interaction effects. The seismic environment was determined from the uniform hazard spectra for several return periods. Based on the results obtained, the safety level of the dam against different seismic actions was determined, and the effectiveness of ambient seismic noise measurements in dynamic characterization and subsequent evaluation of the seismic integrity of urban dams was evaluated.

Keywords: risk, seismic, soil-structure interaction, urban dams

Procedia PDF Downloads 92
4415 GC-MS Analysis of Essential Oil from the Leaves and Fruits of Artemesia Campestris from Algeria

Authors: B. Bakchiche, H. Guenane, M. Bireche, A. Noureddinne, A. Gherib

Abstract:

The chemical composition of the essential oils obtained by hydrodistillation from Artemisia campestris L (family Asteraceae) collected in Djebel Amour (Sahara Atlas, Algeria). Aerial parts were also evaluated by gas chromatography (GC) and gas chromatography coupled to mass spectrometry (GC-MS). The analyses for leaves and fruits of A. campestris resulted in the identification of thirty-one compounds, representing 91.8 % of the total oil and the yields were 0.33% (v/dry weight). The main components were β-pinene and sabinene (25.6% and 17% respectively) followed by α-pinene (9.9%), limonene (6.6 %) and p-cymene (4.1%).

Keywords: essential oil, GC-MS, Artemesia campestris, Algeria

Procedia PDF Downloads 419
4414 Integrating Microcontroller-Based Projects in a Human-Computer Interaction Course

Authors: Miguel Angel Garcia-Ruiz, Pedro Cesar Santana-Mancilla, Laura Sanely Gaytan-Lugo

Abstract:

This paper describes the design and application of a short in-class project conducted in Algoma University’s Human-Computer Interaction (HCI) course taught at the Bachelor of Computer Science. The project was based on the Maker Movement (people using and reusing electronic components and everyday materials to tinker with technology and make interactive applications), where students applied low-cost and easy-to-use electronic components, the Arduino Uno microcontroller board, software tools, and everyday objects. Students collaborated in small teams by completing hands-on activities with them, making an interactive walking cane for blind people. At the end of the course, students filled out a Technology Acceptance Model version 2 (TAM2) questionnaire where they evaluated microcontroller boards’ applications in HCI classes. We also asked them about applying the Maker Movement in HCI classes. Results showed overall students’ positive opinions and response about using microcontroller boards in HCI classes. We strongly suggest that every HCI course should include practical activities related to tinkering with technology such as applying microcontroller boards, where students actively and constructively participate in teams for achieving learning objectives.

Keywords: maker movement, microcontrollers, learning, projects, course, technology acceptance

Procedia PDF Downloads 159
4413 Covalent Binding of Cysteine to a Sol-Gel Material for Cadmium Biosorption from Aqueous Solutions

Authors: Claudiu Marcu, Cristina Paul, Adelina Andelescu, Corneliu Mircea Davidescu, Francisc Péter

Abstract:

Heavy metal pollution has become a more serious environmental problem in the last several decades as a result of its toxicity and insusceptibility to the environment. Methods for removing metal ions from aqueous solution mainly consist of physical, chemical and biochemical procedures. Biosorption is defined as the removal of metal or metalloid species, compounds and particulates from solution by a biological material. Biosorption represents a very attractive method for the removal of toxic metal ions from aqueous effluents because it uses the ability of various biomass to bind the metal ions without the risk of releasing other toxic chemical compounds into the environment. The problem with using biomass or living cells as biosorbents is that their regeneration/reuse is often either impossible or very laborious. One of the most common chelating group found in biosorbents is the thiol group in cysteine. Therefore, we immobilized cysteine using covalent binding using glutaraldehyde as a linker on a synthetic sol-gel support obtained using 3-amino-propyl-trimetoxysilane and trimetoxysilane as precursors. The obtained adsorbents were used for removal of cadmium from aqueous solutions and the removal capacity was investigated in relation to the composition of the sol-gel hybrid composite, the loading of the biomolecule and the physical parameters of the biosorption process. In the same conditions, the bare sol-gel support without cysteine had no Cd removal effect, while the adsorbent with cysteine had an adsorption capacity up to 25.8 mg Cd/g adsorbent at pH 2.0 and 119 mg Cd/g adsorbent at pH 6.6, depending on cadmium concentration and adsorption conditions. We used atomic adsorption spectrometry to assess the cadmium concentration in the samples after the biosorbtion process. The parameters for the Freundlich and Langmuir adsorption isotherms where calculated from plotting the results of the adsorption experiments. The results for cysteine immobilization show a good loading capacity of the sol-gel support which indicates it could be used to immobilize metal binding proteins and by doing so boosting the heavy metal adsorption capacity of the biosorbent.

Keywords: biosorbtion, cadmium, cysteine covalent binding, sol-gel

Procedia PDF Downloads 283
4412 Caged Compounds as Light-Dependent Initiators for Enzyme Catalysis Reactions

Authors: Emma Castiglioni, Nigel Scrutton, Derren Heyes, Alistair Fielding

Abstract:

By using light as trigger, it is possible to study many biological processes, such as the activity of genes, proteins, and other molecules, with precise spatiotemporal control. Caged compounds, where biologically active molecules are generated from an inert precursor upon laser photolysis, offer the potential to initiate such biological reactions with high temporal resolution. As light acts as the trigger for cleaving the protecting group, the ‘caging’ technique provides a number of advantages as it can be intracellular, rapid and controlled in a quantitative manner. We are developing caging strategies to study the catalytic cycle of a number of enzyme systems, such as nitric oxide synthase and ethanolamine ammonia lyase. These include the use of caged substrates, caged electrons and the possibility of caging the enzyme itself. In addition, we are developing a novel freeze-quench instrument to study these reactions, which combines rapid mixing and flashing capabilities. Reaction intermediates will be trapped at low temperatures and will be analysed by using electron paramagnetic resonance (EPR) spectroscopy to identify the involvement of any radical species during catalysis. EPR techniques typically require relatively long measurement times and very often, low temperatures to fully characterise these short-lived species. Therefore, common rapid mixing techniques, such as stopped-flow or quench-flow are not directly suitable. However, the combination of rapid freeze-quench (RFQ) followed by EPR analysis provides the ideal approach to kinetically trap and spectroscopically characterise these transient radical species. In a typical RFQ experiment, two reagent solutions are delivered to the mixer via two syringes driven by a pneumatic actuator or stepper motor. The new mixed solution is then sprayed into a cryogenic liquid or surface, and the frozen sample is then collected and packed into an EPR tube for analysis. The earliest RFQ instrument consisted of a hydraulic ram unit as a drive unit with direct spraying of the sample into a cryogenic liquid (nitrogen, isopentane or petroleum). Improvements to the RFQ technique have arisen from the design of new mixers in order to reduce both the volume and the mixing time. In addition, the cryogenic isopentane bath has been coupled to a filtering system or replaced by spraying the solution onto a surface that is frozen via thermal conductivity with a cryogenic liquid. In our work, we are developing a novel RFQ instrument which combines the freeze-quench technology with flashing capabilities to enable the studies of both thermally-activated and light-activated biological reactions. This instrument also uses a new rotating plate design based on magnetic couplings and removes the need for mechanical motorised rotation, which can otherwise be problematic at cryogenic temperatures.

Keywords: caged compounds, freeze-quench apparatus, photolysis, radicals

Procedia PDF Downloads 195
4411 Phenolic Composition and Antioxidant Activity of Sorbus L. Fruits and Leaves

Authors: Raudone Lina, Raudonis Raimondas, Gaivelyte Kristina, Pukalskas Audrius, Janulis Valdimaras, Viskelis Pranas

Abstract:

Sorbus L. species are widely distributed in the Northern hemisphere and have been used for medicinal purposes in various traditional medicine systems and as food ingredients. Various Sorbus L. raw materials, fruits, leaves, inflorescences, barks, possess diuretic, anti-inflammatory, hypoglycemic, anti-diarrheal and vasoprotective activities. Phenolics, to whom main pharmacological activities are attributed, are compounds of interest due to their notable antioxidant activity. The aim of this study was to determine the antioxidant profiles of fruits and leaves of selected Sorbus L. species (S. anglica, S. aria f. latifolia, S. arranensis, S. aucuparia, S. austriaca, S. caucasica, S. commixta, S. discolor, S. gracilis, S. hostii, S. semi-incisa, S. tianschanica) and to identify the phenolic compounds with potent contribution to antioxidant activity. Twenty two constituents were identified in Sorbus L. species using ultra high performance liquid chromatography coupled to quadruple and time-of-flight mass spectrometers (UPLC–QTOF–MS). Reducing activity of individual constituents was determined using high performance liquid chromatography (HPLC) coupled to post-column FRAP assay. Signicantly greatest trolox equivalent values corresponding up to 45% of contribution to antioxidant activity were assessed for neochlorogenic and chlorogenic acids, which were determined as markers of antioxidant activity in samples of leaves and fruits. Characteristic patterns of antioxidant profiles obtained using HPLC post-column FRAP assay significantly depend on specific Sorbus L. species and raw materials and are suitable for equivalency research of Sorbus L. fruits and leaves. Selecting species and target plant organs with richest phenolic composition and strongly expressed antioxidant power is the first step in further research of standardized extracts.

Keywords: FRAP, antioxidant, phenolic, Sorbus L., chlorogenic acid, neochlorogenic acid

Procedia PDF Downloads 437