Search results for: cell saver
1550 Establishments of an Efficient Platform for Genome Editing in Grapevine
Authors: S. Najafi, E. Bertini, M. Pezzotti, G.B. Tornielli, S. Zenoni
Abstract:
Grapevine is an important agricultural fruit crop plant consumed worldwide and with a key role in the global economy. Grapevine is strongly affected by both biotic and abiotic stresses, which impact grape growth at different stages, such as during plant and berry development and pre- and post-harvest, consequently causing significant economic losses. Recently global warming has propelled the anticipation of the onset of berry ripening, determining the reduction of a grape color and increased volatilization of aroma compounds. Climate change could negatively alter the physiological characteristics of the grape and affect the berry and wine quality. Modern plant breeding can provide tools such as genome editing for improving grape resilience traits while maintaining intact the viticultural and oenological quality characteristics of the genotype. This study aims at developing a platform for genome editing application in grapevine plants with the final goal to improve berry quality, biotic, and abiotic resilience traits. We chose to directly deliver ribonucleoproteins (RNP, preassembled Cas protein and guide RNA) into plant protoplasts, and, from these cell structures, regenerate grapevine plants edited in specific selected genes controlling traits of interest. Edited plants regenerated by somatic embryogenesis from protoplasts will then be sequenced and molecularly characterized. Embryogenic calli of Sultana and Shiraz cultivars were initiated from unopened leaves of in-vitro shoot tip cultures and from stamens, respectively. Leaves were placed on NB2 medium while stamens on callus initiation medium (PIV) medium and incubated in the dark at 28 °C for three months. Viable protoplasts, tested by FDA staining, isolated from embryogenic calli were cultured by disc method at 1*105 protoplasts/ml. Mature well-shaped somatic embryos developed directly in the protoplast culture medium two months later and were transferred in the light into to shooting medium for further growth. Regenerated plants were then transferred to the greenhouse; no phenotypic alterations were observed when compared to non in-vitro cultured plants. The performed experiments allowed to established an efficient protocol of embryogenic calli production, protoplast isolation, and regeneration of the whole plant through somatic embryogenesis in both Sultana and Shiraz. Regenerated plants, through direct somatic embryogenesis deriving from a single cell, avoid the risk of chimerism during the regeneration process, therefore improving the genome editing process. As pre-requisite of genome editing, an efficient method for transfection of protoplast by yellow fluorescent protein (YFP) marker genes was also established and experiments of direct delivery of CRISPR–Cas9 ribonucleoproteins (RNPs) in protoplasts to achieve efficient DNA-free targeted mutations are in progress.Keywords: CRISPR-cas9, plant regeneration, protoplast isolation, Vitis vinifera
Procedia PDF Downloads 1521549 A Low-Power, Low-Noise and High-Gain 58~66 GHz CMOS Receiver Front-End for Short-Range High-Speed Wireless Communications
Authors: Yo-Sheng Lin, Jen-How Lee, Chien-Chin Wang
Abstract:
A 60-GHz receiver front-end using standard 90-nm CMOS technology is reported. The receiver front-end comprises a wideband low-noise amplifier (LNA), and a double-balanced Gilbert cell mixer with a current-reused RF single-to-differential (STD) converter, an LO Marchand balun and a baseband amplifier. The receiver front-end consumes 34.4 mW and achieves LO-RF isolation of 60.7 dB, LO-IF isolation of 45.3 dB and RF-IF isolation of 41.9 dB at RF of 60 GHz and LO of 59.9 GHz. At IF of 0.1 GHz, the receiver front-end achieves maximum conversion gain (CG) of 26.1 dB at RF of 64 GHz and CG of 25.2 dB at RF of 60 GHz. The corresponding 3-dB bandwidth of RF is 7.3 GHz (58.4 GHz to 65.7 GHz). The measured minimum noise figure was 5.6 dB at 64 GHz, one of the best results ever reported for a 60 GHz CMOS receiver front-end. In addition, the measured input 1-dB compression point and input third-order inter-modulation point are -33.1 dBm and -23.3 dBm, respectively, at 60 GHz. These results demonstrate the proposed receiver front-end architecture is very promising for 60 GHz direct-conversion transceiver applications.Keywords: CMOS, 60 GHz, direct-conversion transceiver, LNA, down-conversion mixer, marchand balun, current-reused
Procedia PDF Downloads 4531548 Synthesis, Characterization and Cytotoxic Effect of Eu2O3-doped ZnO Nanostructures
Authors: Otilia R. Vasile, Florina C. Ilie, Irina F. Nicoara, Cristina D. Ghitulica, Roxana Trusca, Ovidiu Oprea, Vasile A. Surdu, Bogdan S. Vasile, Ecaterina Adronescu
Abstract:
In this work ZnO nanostructures (nanopowders and nanostars) have been synthesized via a simple sol-gel method. The used methods for synthesizing the nanostructures involve two steps as follows: (1) precipitation of zinc acetate precursor for the synthesis of ZnO nanopowders and zinc chloride precursor for the synthesis of ZnO nanostars and (2) addition of Eu2O3 in different concentrations (1%, 3%, and 5%) using europium acetate as precursor. Detailed crystalline parameters for each of the synthetized species were analysed using X-ray diffraction. Structural transitions were also discussed. The structure and morphology of the as-prepared ZnO nanopowders and nanostars were investigated by electron microscopy. TEM investigations have shown an average particle size range from 23 to 29 nm and polyhedral and spherical morphology with tendency to form aggregates for nanopowders. For nanostars structures, a star-like morphology could be observed. Cytotoxicity tests on MG-63 cell lines were also performed. Photocatalytic activity of ZnO nanopowders have reached higher values compared to ZnO nanostars.Keywords: cytotoxicity, photocatalytic activity, TEM, ZnO
Procedia PDF Downloads 5641547 Microfluidic Method for Measuring Blood Viscosity
Authors: Eunseop Yeom
Abstract:
Many cardiovascular diseases, such as thrombosis and atherosclerosis, can change biochemical molecules in plasma and red blood cell. These alterations lead to excessive increase of blood viscosity contributing to peripheral vascular diseases. In this study, a simple microfluidic-based method is used to measure blood viscosity. Microfluidic device is composed of two parallel side channels and a bridge channel. To estimate blood viscosity, blood samples and reference fluid are separately delivered into each inlet of two parallel side channels using pumps. An interfacial line between blood samples and reference fluid occurs by blocking the outlet of one side-channel. Since width for this interfacial line is determined by pressure ratio between blood and reference flows, blood viscosity can be estimated by measuring width for this interfacial line. This microfluidic-based method can be used for evaluating variations in the viscosity of animal models with cardiovascular diseases under flow conditions.Keywords: blood viscosity, microfluidic chip, pressure, shear rate
Procedia PDF Downloads 3751546 Capability of a Single Antigen to Induce Both Protective and Disease Enhancing Antibody: An Obstacle in the Creation of Vaccines and Passive Immunotherapies
Authors: Parul Kulshreshtha, Subrata Sinha, Rakesh Bhatnagar
Abstract:
This study was conducted by taking B. anthracis as a model pathogen. On infecting a host, B. anthracis secretes three proteins, namely, protective antigen (PA, 83kDa), edema factor (EF, 89 kDa) and lethal factor (LF, 90 kDa). These three proteins are the components of two anthrax toxins. PA binds to the cell surface receptors, namely, tumor endothelial marker (TEM) 8 and capillary morphogenesis protein (CMG) 2. TEM8 and CMG2 interact with LDL-receptor related protein (LRP) 6 for endocytosis of EF and LF. On entering the cell, EF acts as a calmodulin-dependent adenylate cyclase that causes a prolonged increase of cytosolic cyclic adenosine monophosphate (cAMP). LF is a metalloprotease that cleaves most isoforms of mitogen-activated protein kinase kinases (MAPKK/MEK) close to their N-terminus. By secreting these two toxins, B.anthracis ascertains death of the host. Once the systemic levels of the toxins rise, antibiotics alone cannot save the host. Therefore, toxin-specific inhibitors have to be developed. In this wake, monoclonal antibodies have been developed for the neutralization of toxic effects of anthrax toxins. We created hybridomas by using spleen of mice that were actively immunized with rLFn (recombinant N-terminal domain of lethal factor of B. anthracis) to obtain anti-toxin antibodies. Later on, separate group of mice were immunized with rLFn to obtain a polyclonal control for passive immunization studies of monoclonal antibodies. This led to the identification of one cohort of rLFn-immunized mice that harboured disease-enhancing polyclonal antibodies. At the same time, the monoclonal antibodies from all the hybridomas were being tested. Two hybridomas secreted monoclonal antibodies (H8 and H10) that were cross-reactive with EF (edema factor) and LF (lethal factor), while the other two hybridomas secreted LF-specific antibodies (H7 and H11). The protective efficacy of H7, H8, H10 and H11 was investigated. H7, H8 and H10 were found to be protective. H11 was found to have disease enhancing characteristics in-vitro and in mouse model of challenge with B. anthracis. In this study the disease enhancing character of H11 monoclonal antibody and anti-rLFn polyclonal sera was investigated. Combination of H11 with protective monoclonal antibodies (H8 and H10) reduced its disease enhancing nature both in-vitro and in-vivo. But combination of H11 with LETscFv (an scFv with VH and VL identical to H10 but lacking Fc region) could not abrogate the disease-enhancing character of H11 mAb. Therefore it was concluded that for suppression of disease enhancement, Fc portion was absolutely essential for interaction of H10 with H11. Our study indicates that the protective potential of an antibody depends equally on its idiotype/ antigen specificity and its isotype. A number of monoclonal and engineered antibodies are being explored as immunotherapeutics but it is absolutely essential to characterize each one for their individual and combined protective potential. Although new in the sphere of toxin-based diseases, it is extremely important to characterize the disease-enhancing nature of polyclonal as well as monoclonal antibodies. This is because several anti-viral therapeutics and vaccines have failed in the face of this phenomenon. The passive –immunotherapy thus needs to be well formulated to avoid any contraindications.Keywords: immunotherapy, polyclonal, monoclonal, antibody-dependent disease enhancement
Procedia PDF Downloads 3871545 Ab Initio Study of Structural, Elastic, Electronic and Thermal Properties of Full Heusler
Authors: M. Khalfa, H. Khachai, F. Chiker, K. Bougherara, R. Khenata, G. Murtaza, M. Harmel
Abstract:
A theoretical study of structural, elastic, electronic and thermodynamic properties of Fe2VX, (with X = Al and Ga), were studied by means of the full-relativistic version of the full-potential augmented plane wave plus local orbitals method. For exchange and correlation potential we used both generalized-gradient approximation (GGA) and local-density approximation (LDA). Our calculated ground state properties like as lattice constants, bulk modulus and elastic constants appear more accurate when we employed the GGA rather than the LDA approximation, and these results agree very well with the available experimental and theoretical data. Further, prediction of the thermal effects on some macroscopic properties of Fe2VAl and Fe2VGa are given in this paper using the quasi-harmonic Debye model in which the lattice vibrations are taken into account. We have obtained successfully the variations of the primitive cell volume, volume expansion coefficient, heat capacities and Debye temperature with pressure and temperature in the ranges of 0–40 GPa and 0–1500 K.Keywords: full Heusler, FP-LAPW, electronic properties, thermal properties
Procedia PDF Downloads 4951544 Asymmetric Synthesis and Biological Study of Suberosanes
Authors: Mohammad Kousara, Françoise Dumas, Rama Ibrahim, Joëlle Dubois, Joël Raingeaud
Abstract:
Suberosanes are a small group of marine natural sesquiterpenes discovered since 1996 by Boyd, Sheu and Qi from three gorgonians. Their skeleton was previously found in quadranes produced by the terrestrial fungus Aspergillus terreus. Up to date, eleven suberosanes are described from which (-)-suberosanone and (-)-suberosenol A are reaching the picomolar cytotoxicity level on human solid tumors cell lines. Due to their impressive cytotoxic properties and their limited availability, we undertook an asymmetric synthesis of the most active members of this family in order to get insight into their absolute configurations and their biological properties. The challenge of their synthesis is the regio- and stereoselective elaboration of the compact bridged tricyclic skeleton with up to five all adjacent asymmetric centers, including a central quaternary carbon one. Our strategy is based on an aza-ene-synthesis key step which is regio-and stereo-controlled by the choice of a chiral amine enantiomer. it approach is concise and flexible, the enantiopur ABC tricyclic intermediate that have been synthesized being the common precursor of suberosanes.Keywords: suberosanes, asymmetric synthesis, sesquiterpenes, quadranes
Procedia PDF Downloads 941543 3D Label-Free Bioimaging of Native Tissue with Selective Plane Illumination Optical Microscopy
Authors: Jing Zhang, Yvonne Reinwald, Nick Poulson, Alicia El Haj, Chung See, Mike Somekh, Melissa Mather
Abstract:
Biomedical imaging of native tissue using light offers the potential to obtain excellent structural and functional information in a non-invasive manner with good temporal resolution. Image contrast can be derived from intrinsic absorption, fluorescence, or scatter, or through the use of extrinsic contrast. A major challenge in applying optical microscopy to in vivo tissue imaging is the effects of light attenuation which limits light penetration depth and achievable imaging resolution. Recently Selective Plane Illumination Microscopy (SPIM) has been used to map the 3D distribution of fluorophores dispersed in biological structures. In this approach, a focused sheet of light is used to illuminate the sample from the side to excite fluorophores within the sample of interest. Images are formed based on detection of fluorescence emission orthogonal to the illumination axis. By scanning the sample along the detection axis and acquiring a stack of images, 3D volumes can be obtained. The combination of rapid image acquisition speeds with the low photon dose to samples optical sectioning provides SPIM is an attractive approach for imaging biological samples in 3D. To date all implementations of SPIM rely on the use of fluorescence reporters be that endogenous or exogenous. This approach has the disadvantage that in the case of exogenous probes the specimens are altered from their native stage rendering them unsuitable for in vivo studies and in general fluorescence emission is weak and transient. Here we present for the first time to our knowledge a label-free implementation of SPIM that has downstream applications in the clinical setting. The experimental set up used in this work incorporates both label-free and fluorescent illumination arms in addition to a high specification camera that can be partitioned for simultaneous imaging of both fluorescent emission and scattered light from intrinsic sources of optical contrast in the sample being studied. This work first involved calibration of the imaging system and validation of the label-free method with well characterised fluorescent microbeads embedded in agarose gel. 3D constructs of mammalian cells cultured in agarose gel with varying cell concentrations were then imaged. A time course study to track cell proliferation in the 3D construct was also carried out and finally a native tissue sample was imaged. For each sample multiple images were obtained by scanning the sample along the axis of detection and 3D maps reconstructed. The results obtained validated label-free SPIM as a viable approach for imaging cells in a 3D gel construct and native tissue. This technique has the potential use in a near-patient environment that can provide results quickly and be implemented in an easy to use manner to provide more information with improved spatial resolution and depth penetration than current approaches.Keywords: bioimaging, optics, selective plane illumination microscopy, tissue imaging
Procedia PDF Downloads 2511542 Detection and Quantification of Viable but Not Culturable Vibrio Parahaemolyticus in Frozen Bivalve Molluscs
Authors: Eleonora Di Salvo, Antonio Panebianco, Graziella Ziino
Abstract:
Background: Vibrio parahaemolyticus is a human pathogen that is widely distributed in marine environments. It is frequently isolated from raw seafood, particularly shellfish. Consumption of raw or undercooked seafood contaminated with V. parahaemolyticus may lead to acute gastroenteritis. Vibrio spp. has excellent resistance to low temperatures so it can be found in frozen products for a long time. Recently, the viable but non-culturable state (VBNC) of bacteria has attracted great attention, and more than 85 species of bacteria have been demonstrated to be capable of entering this state. VBNC cells cannot grow in conventional culture medium but are viable and maintain metabolic activity, which may constitute an unrecognized source of food contamination and infection. Also V. parahaemolyticus could exist in VBNC state under nutrient starvation or low-temperature conditions. Aim: The aim of the present study was to optimize methods and investigate V. parahaemolyticus VBNC cells and their presence in frozen bivalve molluscs, regularly marketed. Materials and Methods: propidium monoazide (PMA) was integrated with real-time polymerase chain reaction (qPCR) targeting the tl gene to detect and quantify V. parahaemolyticus in the VBNC state. PMA-qPCR resulted highly specific to V. parahaemolyticus with a limit of detection (LOD) of 10-1 log CFU/mL in pure bacterial culture. A standard curve for V. parahaemolyticus cell concentrations was established with the correlation coefficient of 0.9999 at the linear range of 1.0 to 8.0 log CFU/mL. A total of 77 samples of frozen bivalve molluscs (35 mussels; 42 clams) were subsequently subjected to the qualitative (on alkaline phosphate buffer solution) and quantitative research of V. parahaemolyticus on thiosulfate-citrate-bile salts-sucrose (TCBS) agar (DIFCO) NaCl 2.5%, and incubation at 30°C for 24-48 hours. Real-time PCR was conducted on homogenate samples, in duplicate, with and without propidium monoazide (PMA) dye, and exposed for 45 min under halogen lights (650 W). Total DNA was extracted from cell suspension in homogenate samples according to bolliture protocol. The Real-time PCR was conducted with species-specific primers for V. parahaemolitycus. The RT-PCR was performed in a final volume of 20 µL, containing 10 µL of SYBR Green Mixture (Applied Biosystems), 2 µL of template DNA, 2 µL of each primer (final concentration 0.6 mM), and H2O 4 µL. The qPCR was carried out on CFX96 TouchTM (Bio-Rad, USA). Results: All samples were negative both to the quantitative and qualitative detection of V. parahaemolyticus by the classical culturing technique. The PMA-qPCR let us individuating VBNC V. parahaemolyticus in the 20,78% of the samples evaluated with a value between the Log 10-1 and Log 10-3 CFU/g. Only clams samples were positive for PMA-qPCR detection. Conclusion: The present research is the first evaluating PMA-qPCR assay for detection of VBNC V. parahaemolyticus in bivalve molluscs samples, and the used method was applicable to the rapid control of marketed bivalve molluscs. We strongly recommend to use of PMA-qPCR in order to identify VBNC forms, undetectable by the classic microbiological methods. A precise knowledge of the V.parahaemolyticus in a VBNC form is fundamental for the correct risk assessment not only in bivalve molluscs but also in other seafood.Keywords: food safety, frozen bivalve molluscs, PMA dye, Real-time PCR, VBNC state, Vibrio parahaemolyticus
Procedia PDF Downloads 1401541 Control of Stability for PV and Battery Hybrid System in Partial Shading
Authors: Weiying Wang, Qi Li, Huiwen Deng, Weirong Chen
Abstract:
The abrupt light change and uneven illumination will make the PV system get rid of constant output power, which will affect the efficiency of the grid connected inverter as well as the stability of the system. To solve this problem, this paper presents a strategy to control the stability of photovoltaic power system under the condition of partial shading of PV array, leading to constant power output, improving the capacity of resisting interferences. Firstly, a photovoltaic cell model considering the partial shading is established, and the backtracking search algorithm is used as the maximum power point to track algorithm under complex illumination. Then, the energy storage system based on the constant power control strategy is used to achieve constant power output. Finally, the effectiveness and correctness of the proposed control method are verified by the joint simulation of MATLAB/Simulink and RTLAB simulation platform.Keywords: backtracking search algorithm, constant power control, hybrid system, partial shading, stability
Procedia PDF Downloads 2981540 Screening Active Components in YPFS for Regulating Initiative Key Factors in Allergic Inflammation
Authors: Dandan Shen, Hui-zhu Wang, Xi Yu, LiLi Gui, Xiao Wei, Xiao-yan Jiang, Da-wei Wang, Min Hong
Abstract:
Yu-ping-feng-san (YPFS) is a clinical medicine for asthma and other allergic diseases, but the mechanism of YPFS on relapse of allergy is unclear. Currently, people come to realize the epithelial cells(EC) play a key role in stimulating and regulating local immune response. The study of thymic stromal lymphopoietin(TSLP derived from EC provides an important evidence that the EC can regulate immune response to stimulate allergic response. In this study, we observed the effect of YPFS on TSLP in vivo and in vitro. We established a method by using bronchial epithelial cells (16HBE) for screening potential bioactive components by HPLC-MS in YPFS and then analyzed the components in serum containing YPFS by UPLC-MS. The results showed that YPFS could decrease TSLP protein level in OVA-sensitized mice and 16HBE cells. Five components combing with the 16HBE cells were both detected in the serum.Keywords: TSLP, bronchial epithelial cells, cell-binding, drug-containing serum
Procedia PDF Downloads 5151539 Toxicity of Solenstemma Argel (Hargal ) on Nubian Goats
Authors: Amna B. Medani, M. A. Elbadwi Samia, Hassan A. Khalid
Abstract:
In our study, nine Nubian goat kids were obtained, allotted into three groups, and healthily adapted in pens within the premises of the Veterinary Teaching Hospital, University of Khartoum to be given the oral doses of the dried herb shoots at daily doses of 1 and 5 gm/kg/day with drinking water, while the kids of the control group were left undosed. All goats were slaughtered,if not died, after 35 days. S. argel at the given doses caused signs of arched posture, ruffled hair, shivering and paralysis of limbs. On post mortem, lesions were seen to be hepatic fatty changes, renal necrosis, congested lungs and inflamed intestines. Serum chemistry investigations revealed significant increase (P< 0.05-0.01) in the activities of ALP(alkaline phosphates) and AST( aspartate-aminotransferase) in goats dosed with 5 gm /kg/ day. Also observed were significant increases in inorganic phosphorus and urea concentrations (P < 0.05-0.01) in both dosed goat groups. .Other investigations including the activity of GGT( gamma glutamyltransferase), creatinine, calcium, total protein and albumin illustrated no significant difference from that of the undosed controls. On haematological evaluation , the goat kids dosed with 5 gm/kg/dayshowed a decrease in haemoglobin concentration and red blood cells count of (P < 0.05-0.01).Both groups of dosed goats showed a higher packed cell volume values of (P < 0.05) when compared to the control goats .Mean corpuscular haemoglobin values were not different from those of the control kids. S. argel at the given doses caused signs of arched posture, ruffled hair, shivering and paralysis of limbs. On post mortem, lesions were seen to be hepatic fatty changes, renal necrosis, congested lungs and inflamed intestines. Serum chemistry investigations revealed significant increase (P < 0.05-0.01) in the activities of ALP(alkaline phosphates) and AST( aspartate-aminotransferase) in goats dosed with 5 gm /kg/ day. Also observed were significant increases in inorganic phosphorus and urea concentrations (P < 0.05-0.01) in both dosed goat groups. .Other investigations including the activity of GGT( gamma-glutamyltransferase), creatinine, calcium, total protein and albumin illustrated no significant difference from that of the undosed controls. calcium, total protein and albumin illustrated no significant difference from that of the undosed controls. On haematological evaluation , the goat kids dosed with 5 gm/kg/dayshowed a decrease in haemoglobin concentration and red blood cells count of (P < 0.05-0.01).Both groups of dosed goats showed a higher packed cell volume values of (P < 0.05) when compared to the control goats .Mean corpuscular haemoglobin values were not different from those of the control kids. Data obtained were then discussed to find S. argel irritable to intestines , toxic to the kidney and liver and a haematological mild toxin.Suggestions for future were forwarded.Keywords: hargal, nubian goats, solenstemma argel, toxicity
Procedia PDF Downloads 3221538 Contraction and Membrane Potential of C2C12 with GTXs
Authors: Bayan Almofty, Yuto Yamaki, Tadamasa Terai, Sadahito Uto
Abstract:
Culture techniques of skeletal muscle cells are advanced in the field of regenerative medicine and applied research of cultured muscle. As applied research of cultured muscle, myopathy (muscles disease) treatment is expected and development bio of actuator is also expected in biomedical engineering. Grayanotoxins (GTXs) is known as neurotoxins that enhance the permeability of cell membrane for Na ions. Grayanotoxins are extracted from a famous Pieris japonica and Ericaceae as well as a phytotoxin. In this study, we investigated the effect of GTXs on muscle cells (C2C12) contraction and membrane potential. Contraction of myotubes is induced by applied external electrical stimulation. Contraction and membrane potential change of skeletal muscle cells are induced by injection of current. We, therefore, concluded that effect of Grayanotoxins on contraction and membrane potential of C2C12 relate to acute toxicity of GTXs.Keywords: skeletal muscle cells C2C12, grayanotoxins, contraction, membrane potential, acute toxicity, pytotoxin, motubes
Procedia PDF Downloads 5061537 Synthesis of Quinazoline Derivatives as Selective Inhibitors of Cyclooxygenase-1 Enzyme
Authors: Marcela Dvorakova, Lenka Langhansova, Premysl Landa
Abstract:
A series of quinazoline derivatives bearing aromatic rings in 2- and 4-positions were prepared and tested for their biological activity. Firstly, the compounds were evaluated for their potential to inhibit various kinases, such as autophagy activating kinase ULK1, 3-Phosphoinositide-dependent kinase 1, and TANK-binding kinase 1. None of the compounds displayed any activity on these kinases. Secondly, the compounds were tested for their anti-inflammatory activity expressed as cyclooxygenase (COX) isoforms and 5-lipoxygenase (5-LOX) inhibition. Three of the compounds showed significant selectivity towards COX-1 isoform (COX-2/COX-1 SI = 20-30). They inhibited COX-1 in a single-digit µM range. There was also one compound that exhibited inhibitory activity towards all three tested enzymes in µM range (IC50COX-1 = 1.9 µM; IC50COX-2 and 5-LOX = 10.1µM. COX-1 inhibition was until recently considered undesirable due to COX-1 constitutive expression in most cell types and tissues. Thus, there are not many compounds known with selective COX-1 activity. However, it is now believed that COX-1 plays an important role in the pathophysiology of several acute and chronic disorders, including cancer or neurodegenerative diseases. Thus, the discovery of effective COX-1 selective inhibitors is desirable and important.Keywords: cyclooxygenases, kinases, lipoxygenases, quinazolines
Procedia PDF Downloads 1371536 Biofilm Formation Due to the Proteome Changes Of Enterococcus Faecium in Response to Sub-Mic of Gentamicin
Authors: Amin Abbasi, Mahdi Asghari Ozma
Abstract:
Background and Objective:Enterococcus faecium is a normal flora of the human gastrointestinal tract that causes infection in the host body under conditions such as biofilm formation, in which the use of antibiotics causes changes in these pathogenic mechanisms. In this study, we aimed to evaluate comprehensively the changes in E.faecium when exposed to sub-MIC of the gentamicin,especiallythe biofilm formation rate. Materials and Methods: For this study, the keywords "Enterococcus faecium ", "Biofilm", and "Gentamicin" in the databases PubMed, Google Scholar, Sid, and MagIran between 2015 and 2021 were searched, and 14 articles were chosen, studied, and analyzed. Results: Gentamicin significantly had increased biofilm formation in most of the isolates in the studies. Increased expression of the genes (efaA and esp) and proteins involved in biofilm formation and decreased expression of the genes (gelE and cylA) involved in spreading and proteins involved in metabolism and cell division in E.faecium were the most significant cause of the biofilm formation, which were increased in sub-MIC gentamicin-treated situation. Conclusion: Inadequate use of gentamicin intensify biofilm formation of E.faecium, which can make the treatment of infections caused by this bacterium difficult.Keywords: biofilm, enterococcus faecium, gentamicin, proteome
Procedia PDF Downloads 1121535 Fabrication and Properties of Al2O3/Si Quantum Well-Structured Silicon Solar Cells
Authors: Kwang-Ho Kim, Kwan-Hong Min, Pyungwoo Jang, Chisup Jung, Kyu Seomoon
Abstract:
By restricting the dimensions of silicon to less than Bohr radius of bulk crystalline silicon (∼5 nm), quantum confinement causes its effective bandgap to increase. Therefore, silicon quantum wells (QWs) using these quantum phenomena could be a good candidate to achieve high performance silicon solar cells. The Al2O3/Si QW structures were fabricated by using the successive deposition technique, as a quantum confinement device to increase the effective energy bandgap and passivation effect in Si surface for the 3rd generation solar cell applications. In Si/Al2O3 QWs, the thicknesses of Si layers and Al2O3 layers were varied between 1 to 5 nm, respectively. The roughness of deposited Si on Al2O3 was less than 4 Å in the thickness of 2 nm. By using the Al2O3/Si QW structures on Si surfaces, the lifetime measured by u-PCD technique increased as a result of passivated surface effects. The discussion about the other properties such as electrical and optical properties of the QWs structures as well as the fabricated solar cells will be presented in this paper.Keywords: Al2O3/Si quantum well, quantum confinement, solar cells, third generation, successive deposition technique
Procedia PDF Downloads 3401534 Active Learning in Engineering Courses Using Excel Spreadsheet
Authors: Promothes Saha
Abstract:
Recently, transportation engineering industry members at the study university showed concern that students lacked the skills needed to solve real-world engineering problems using spreadsheet data analysis. In response to the concerns shown by industry members, this study investigated how to engage students in a better way by incorporating spreadsheet analysis during class - also, help them learn the course topics. Helping students link theoretical knowledge to real-world problems can be a challenge. In this effort, in-class activities and worksheets were redesigned to integrate with Excel to solve example problems using built-in tools including cell referencing, equations, data analysis tool pack, solver tool, conditional formatting, charts, etc. The effectiveness of this technique was investigated using students’ evaluations of the course, enrollment data, and students’ comments. Based on the data of those criteria, it is evident that the spreadsheet activities may increase student learning.Keywords: civil, engineering, active learning, transportation
Procedia PDF Downloads 1391533 Mobile Microscope for the Detection of Pathogenic Cells Using Image Processing
Authors: P. S. Surya Meghana, K. Lingeshwaran, C. Kannan, V. Raghavendran, C. Priya
Abstract:
One of the most basic and powerful tools in all of science and medicine is the light microscope, the fundamental device for laboratory as well as research purposes. With the improving technology, the need for portable, economic and user-friendly instruments is in high demand. The conventional microscope fails to live up to the emerging trend. Also, adequate access to healthcare is not widely available, especially in developing countries. The most basic step towards the curing of a malady is the diagnosis of the disease itself. The main aim of this paper is to diagnose Malaria with the most common device, cell phones, which prove to be the immediate solution for most of the modern day needs with the development of wireless infrastructure allowing to compute and communicate on the move. This opened up the opportunity to develop novel imaging, sensing, and diagnostics platforms using mobile phones as an underlying platform to address the global demand for accurate, sensitive, cost-effective, and field-portable measurement devices for use in remote and resource-limited settings around the world.Keywords: cellular, hand-held, health care, image processing, malarial parasites, microscope
Procedia PDF Downloads 2681532 Visual Detection of Escherichia coli (E. coli) through Formation of Beads Aggregation in Capillary Tube by Rolling Circle Amplification
Authors: Bo Ram Choi, Ji Su Kim, Juyeon Cho, Hyukjin Lee
Abstract:
Food contaminated by bacteria (E.coli), causes food poisoning, which occurs to many patients worldwide annually. We have introduced an application of rolling circle amplification (RCA) as a versatile biosensor and developed a diagnostic platform composed of capillary tube and microbeads for rapid and easy detection of Escherichia coli (E. coli). When specific mRNA of E.coli is extracted from cell lysis, rolling circle amplification (RCA) of DNA template can be achieved and can be visualized by beads aggregation in capillary tube. In contrast, if there is no bacterial pathogen in sample, no beads aggregation can be seen. This assay is possible to detect visually target gene without specific equipment. It is likely to the development of a genetic kit for point of care testing (POCT) that can detect target gene using microbeads.Keywords: rolling circle amplification (RCA), Escherichia coli (E. coli), point of care testing (POCT), beads aggregation, capillary tube
Procedia PDF Downloads 3701531 The Five Aggregates in Buddhism and Natural Sciences: A Revolutionary Perspective of Nature
Authors: Choo Fatt Foo
Abstract:
The Five Aggregates is core to Buddhism teaching. According to Buddhism, human beings and all sentient beings are made up of nothing but the Five Aggregates. If that is the case, the Five Aggregates must be found in all natural sciences. So far, there has not been any systematic connection between the Five Aggregates and natural sciences. This study aims at identifying traces of the Five Aggregates in various levels of natural sciences and pointing possible directions for future research. The following areas are briefly explored to identify the connection with the Five Aggregates: physics, chemistry, organic chemistry, DNA, cell, and human body and brain. Traces of the Five Aggregates should be found in each level of this hierarchy of natural sciences for human and sentient beings to be said to be made up of the Five Aggregates. This study proposes a hierarchical structure of nature cutting every level with the Five Aggregates and the Four Great Elements as its basis. The structure proposed by this study would revolutionize how we look at nature. Hopefully, better understanding of sciences in this manner will steer the application of scientific methods and technology towards a brighter future with compassion and tolerance.Keywords: the five aggregates, Buddhism, four great elements, physics, calabi-yau manifold
Procedia PDF Downloads 1941530 Collagen/Hydroxyapatite Compositions Doped with Transitional Metals for Bone Tissue Engineering Applications
Authors: D. Ficai, A. Ficai, D. Gudovan, I. A. Gudovan, I. Ardelean, R. Trusca, E. Andronescu, V. Mitran, A. Cimpean
Abstract:
In the last years, scientists struggled hardly to mimic bone structures to develop implants and biostructures which present higher biocompatibility and reduced rejection rate. One way to obtain this goal is to use similar materials as that of bone, namely collagen/hydroxyapatite composite materials. However, it is very important to tailor both compositions but also the microstructure of the bone that would ensure both the optimal osteointegartion and the mechanical properties required by the application. In this study, new collagen/hydroxyapatites composite materials doped with Cu, Li, Mn, Zn were successfully prepared. The synthesis method is described below: weight the Ca(OH)₂ mass, i.e., 7,3067g, and ZnCl₂ (0.134g), CuSO₄ (0.159g), LiCO₃ (0.133g), MnCl₂.4H₂O (0.1971g), and suspend in 100ml distilled water under magnetic stirring. The solution thus obtained is added a solution of NaH₂PO₄*H2O (8.247g dissolved in 50ml distilled water) under slow dropping of 1 ml/min followed by adjusting the pH to 9.5 with HCl and finally filter and wash until neutral pH. The as-obtained slurry was dried in the oven at 80°C and then calcined at 600°C in order to ensure a proper purification of the final product of organic phases, also inducing a proper sterilization of the mixture before insertion into the collagen matrix. The collagen/hydroxyapatite composite materials are tailored from morphological point of view to optimize their biocompatibility and bio-integration against mechanical properties whereas the addition of the dopants is aimed to improve the biological activity of the samples. The addition of transitional metals can improve the biocompatibility and especially the osteoblasts adhesion (Mn²⁺) or to induce slightly better osteoblast differentiation of the osteoblast, Zn²⁺ being a cofactor for many enzymes including those responsible for cell differentiation. If the amount is too high, the final material can become toxic and lose all of its biocompatibility. In order to achieve a good biocompatibility and not reach the cytotoxic effect, the amount of transitional metals added has to be maintained at low levels (0.5% molar). The amount of transitional metals entering into the elemental cell of HA will be verified using inductively-coupled plasma mass spectrometric system. This highly sensitive technique is necessary, because, at such low levels of transitional metals, the difference between biocompatible and cytotoxic is a very thin line, thus requiring proper and thorough investigation using a precise technique. In order to determine the structure and morphology of the obtained composite materials, IR spectroscopy, X-Ray diffraction (XRD), scanning electron microscopy (SEM), and Energy Dispersive X-Ray Spectrometry (EDS) were used. Acknowledgment: The present work was possible due to the EU-funding grant POSCCE-A2O2.2.1-2013-1, Project No. 638/12.03.2014, code SMIS-CSNR 48652. The financial contribution received from the national project “Biomimetic porous structures obtained by 3D printing developed for bone tissue engineering (BIOGRAFTPRINT), No. 127PED/2017 is also highly acknowledged.Keywords: collagen, composite materials, hydroxyapatite, bone tissue engineering
Procedia PDF Downloads 2091529 Numerical Model for Investigation of Recombination Mechanisms in Graphene-Bonded Perovskite Solar Cells
Authors: Amir Sharifi Miavaghi
Abstract:
It is believed recombination mechnisms in graphene-bonded perovskite solar cells based on numerical model in which doped-graphene structures are employed as anode/cathode bonding semiconductor. Moreover, the dark-light current density-voltage density-voltage curves are investigated by regression analysis. Loss mechanisms such as back contact barrier, deep surface defect in the adsorbent layer is determined by adapting the simulated cell performance to the measurements using the differential evolution of the global optimization algorithm. The performance of the cell in the connection process includes J-V curves that are examined at different temperatures and open circuit voltage (V) under different light intensities as a function of temperature. Based on the proposed numerical model and the acquired loss mechanisms, our approach can be used to improve the efficiency of the solar cell further. Due to the high demand for alternative energy sources, solar cells are good alternatives for energy storage using the photovoltaic phenomenon.Keywords: numerical model, recombination mechanism, graphen, perovskite solarcell
Procedia PDF Downloads 721528 Ultrasound Disintegration as a Potential Method for the Pre-Treatment of Virginia Fanpetals (Sida hermaphrodita) Biomass before Methane Fermentation Process
Authors: Marcin Dębowski, Marcin Zieliński, Mirosław Krzemieniewski
Abstract:
As methane fermentation is a complex series of successive biochemical transformations, its subsequent stages are determined, to a various extent, by physical and chemical factors. A specific state of equilibrium is being settled in the functioning fermentation system between environmental conditions and the rate of biochemical reactions and products of successive transformations. In the case of physical factors that influence the effectiveness of methane fermentation transformations, the key significance is ascribed to temperature and intensity of biomass agitation. Among the chemical factors, significant are pH value, type, and availability of the culture medium (to put it simply: the C/N ratio) as well as the presence of toxic substances. One of the important elements which influence the effectiveness of methane fermentation is the pre-treatment of organic substrates and the mode in which the organic matter is made available to anaerobes. Out of all known and described methods for organic substrate pre-treatment before methane fermentation process, the ultrasound disintegration is one of the most interesting technologies. Investigations undertaken on the ultrasound field and the use of installations operating on the existing systems result principally from very wide and universal technological possibilities offered by the sonication process. This physical factor may induce deep physicochemical changes in ultrasonicated substrates that are highly beneficial from the viewpoint of methane fermentation processes. In this case, special role is ascribed to disintegration of biomass that is further subjected to methane fermentation. Once cell walls are damaged, cytoplasm and cellular enzymes are released. The released substances – either in dissolved or colloidal form – are immediately available to anaerobic bacteria for biodegradation. To ensure the maximal release of organic matter from dead biomass cells, disintegration processes are aimed to achieve particle size below 50 μm. It has been demonstrated in many research works and in systems operating in the technical scale that immediately after substrate supersonication the content of organic matter (characterized by COD, BOD5 and TOC indices) was increasing in the dissolved phase of sedimentation water. This phenomenon points to the immediate sonolysis of solid substances contained in the biomass and to the release of cell material, and consequently to the intensification of the hydrolytic phase of fermentation. It results in a significant reduction of fermentation time and increased effectiveness of production of gaseous metabolites of anaerobic bacteria. Because disintegration of Virginia fanpetals biomass via ultrasounds applied in order to intensify its conversion is a novel technique, it is often underestimated by exploiters of agri-biogas works. It has, however, many advantages that have a direct impact on its technological and economical superiority over thus far applied methods of biomass conversion. As for now, ultrasound disintegrators for biomass conversion are not produced on the mass-scale, but by specialized groups in scientific or R&D centers. Therefore, their quality and effectiveness are to a large extent determined by their manufacturers’ knowledge and skills in the fields of acoustics and electronic engineering.Keywords: ultrasound disintegration, biomass, methane fermentation, biogas, Virginia fanpetals
Procedia PDF Downloads 3701527 An AFM Approach of RBC Micro and Nanoscale Topographic Features During Storage
Authors: K. Santacruz-Gomez, E. Silva-Campa, S. Álvarez-García, V. Mata-Haro, D. Soto-Puebla, M. Pedroza-Montero
Abstract:
Blood gamma irradiation is the only available method to prevent transfusion-associated graft versus host disease (TA-GVHD). However, when blood is irradiated, determine blood shelf time is crucial. Non-irradiated blood has a self-time from 21 to 35 days when is preserved with an anticoagulated solution and stored at 4°C. During their storage, red blood cells (RBC) undergo a series of biochemical, biomechanical and molecular changes involving what is known as storage lesion (SL). SL include loss of structural integrity of RBC, a decrease of 2,3-diphosphatidylglyceric acid levels, and an increase of both ion potassium concentration and hemoglobin (Hb). On the other hand, Atomic force Microscopy (AFM) represents a versatile tool for a nano-scale high-resolution topographic analysis in biological systems. In order to evaluate SL in irradiated and non-irradiated blood, RBC topography and morphometric parameters were obtained from an AFM XE-BIO system. Cell viability was followed using flow cytometry. Our results showed that early markers as nanoscale roughness, allow us to evaluate blood quality since another perspective.Keywords: AFM, blood γ-irradiation, roughness, storage lesion
Procedia PDF Downloads 5331526 The Effect of Pre-Cracks on Structural Strength of the Nextel Fibers: A Multiscale Modeling Approach
Authors: Seyed Mohammad Mahdi Zamani, Kamran Behdinan
Abstract:
In this study, a multiscale framework is performed to model the strength of Nextel fibers in presence of an atomistic scale pre-crack at finite temperatures. The bridging cell method (BCM) is the multiscale technique applied in this study, which decomposes the system into the atomistic, bridging and continuum domains; solves the whole system in a finite element framework; and incorporates temperature dependent calculations. Since Nextel is known to be structurally stable and retain 70% of its initial strength up to 1100°C; simulations are conducted at both of the room temperatures, 25°C, and fire temperatures, 1200°C. Two cases are modeled for a pre-crack present in either phases of alumina or mullite of the Nextel structure. The materials’ response is studied with respect to deformation behavior and ultimate tensile strength. Results show different crack growth trends for the two cases, and as the temperature increases, the crack growth resistance and material’s strength decrease.Keywords: Nextel fibers, multiscale modeling, pre-crack, ultimate tensile strength
Procedia PDF Downloads 4211525 Hafnium Doped Zno Nanostructures: An Eco-Friendly Synthesis for Optoelectronic Applications
Authors: Mohamed Achehboune, Mohammed Khenfouch, Issam Boukhoubza, Bakang Mothudi, Izeddine Zorkani, Anouar Jorio
Abstract:
Zinc Oxide (ZnO) nanostructures have been attracting growing interest in recent years; their optical and electrical properties make them useful as attractive and promising materials for optoelectronic applications. In this study, pure and Hafnium doped ZnO nanostructures were synthesized using a green processing method. The structural, optical and electrical properties of samples were investigated structural and optical spectroscopies and electrical measurements. The synthesis and chemical composition of pure and Hafnium doped ZnO were confirmed by SEM observation. The XRD studies of Hafnium doped ZnO demonstrate the formation of wurtzite structure with preferred c-axis orientation. Moreover, the optical and electrical properties of doped material have improved after the doping process. The experimental results obtained for our material show that Hf doped ZnO nanostructures could be a promising material in optoelectronic applications such as photovoltaic cell and light emitting diode devices.Keywords: green synthesis, hafnium-doped-zinc oxide, nanostructures, optoelectronic
Procedia PDF Downloads 2741524 Simulation of Red Blood Cells in Complex Micro-Tubes
Authors: Ting Ye, Nhan Phan-Thien, Chwee Teck Lim, Lina Peng, Huixin Shi
Abstract:
In biofluid flow systems, often the flow problems of fluids of complex structures, such as the flow of red blood cells (RBCs) through complex capillary vessels, need to be considered. In this paper, we aim to apply a particle-based method, Smoothed Dissipative Particle Dynamics (SDPD), to simulate the motion and deformation of RBCs in complex micro-tubes. We first present the theoretical models, including SDPD model, RBC-fluid interaction model, RBC deformation model, RBC aggregation model, and boundary treatment model. After that, we show the verification and validation of these models, by comparing our numerical results with the theoretical, experimental and previously-published numerical results. Finally, we provide some simulation cases, such as the motion and deformation of RBCs in rectangular, cylinder, curved, bifurcated, and constricted micro-tubes, respectively.Keywords: aggregation, deformation, red blood cell, smoothed dissipative particle dynamics
Procedia PDF Downloads 1751523 Role of GM1 in the Interaction between Amyloid Prefibrillar Oligomers of Salmon Calcitonin and Model Membranes
Authors: Cristiano Giordani, Marco Diociaiuti, Cecilia Bombelli, Laura Zanetti-Polzi, Marcello Belfiore, Raoul Fioravanti, Gianfranco Macchia
Abstract:
We investigated induced functional effects by evaluating Ca2+-influx in liposomes and cell viability in HT22-DIFF neurons. Only solutions rich in unstructured Prefibrillar-Oligomers (PFOs) were able, in the presence of Monosialoganglioside-GM1 (GM1), to induce Ca2+-influx and were also neurotoxic, suggesting a correlation between the two phenomena. Thus, in the presence of GM1, we investigated the protein conformation and liposome modification due to the interaction. Circular Dichroism showed that GM1 fostered the formation of β-structures and Energy Filtered-Transmission Electron Microscopy that PFOs formed “amyloid-channels” as reported for Aβ. We speculate that electrostatic forces occurring between the positive PFOs and negative GM1 drive the initial binding, while the hydrophobic profile of the flexible PFO is responsible for the subsequent pore formation. Conversely, the rigid β-structured mature/fibers (MFs) and proto-fibers (PFs) were unable to induce membrane damage and Ca2+- influx.Keywords: amyloid proteins, neurotoxicity, lipid-rafts, GM1
Procedia PDF Downloads 1911522 Harnessing Nature's Fury: Hyptis Suaveolens Loaded Bioactive Liposome for Photothermal Therapy of Lung Cancer
Authors: Sajmina Khatun, Monika Pebam, Aravind Kumar Rengan
Abstract:
Photothermal therapy, a subset of nanomedicine, takes advantage of light-absorbing agents to generate localized heat, selectively eradicating cancer cells. This innovative approach minimizes damage to healthy tissues and offers a promising avenue for targeted cancer treatment. Unlike conventional therapies, photothermal therapy harnesses the power of light to combat malignancies precisely and effectively, showcasing its potential to revolutionize cancer treatment paradigms. The combined strengths of nanomedicine and photothermal therapy signify a transformative shift toward more effective, targeted, and tolerable cancer treatments in the medical landscape. Utilizing natural products becomes instrumental in formulating diverse bioactive medications owing to their various pharmacological properties attributed to the existence of phenolic structures, triterpenoids, and similar compounds. Hyptis suaveolens, commonly known as pignut, stands as an aromatic herb within the Lamiaceae family and represents a valuable therapeutic plant. Flourishing in swamps and alongside tropical and subtropical roadsides, these noxious weeds impede the development of adjacent plants. Hyptis suaveolens ranks among the most globally distributed alien invasive species. The present investigation revealed that a versatile, biodegradable liposome nanosystem (HIL NPs), incorporating bioactive molecules from Hyptis suaveolens, exhibits effective bioavailability to cancer cells, enabling tumor ablation upon near-infrared (NIR) laser exposure. The components within the nanosystem, specifically the bioactive molecules from Hyptis, function as anticancer agents, aiding in the photothermal ablation of highly metastatic lung cancer cells. Despite being a prolific weed impeding neighboring plant growth, Hyptis suaveolens showcases therapeutic benefits through its bioactive compounds. The obtained HIL NPs, characterized as a photothermally active liposome nanosystem, demonstrate a pronounced fluorescence absorption peak in the NIR range and achieve a high photothermal conversion efficiency under NIR laser irradiation. Transmission electron microscopy (TEM) and particle size analysis reveal that HIL NPs possess a spherical shape with a size of 141 ± 30 nm. Moreover, in vitro assessments of HIL NPs against lung cancer cell lines (A549) indicate effective anticancer activity through a combined cytotoxic effect and hyperthermia. Tumor ablation is facilitated by apoptosis induced by the overexpression of ɣ-H2AX, arresting cancer cell proliferation. Consequently, the multifunctional and biodegradable nanosystem (HIL NPs), incorporating bioactive compounds from Hyptis, provides valuable perspectives for developing an innovative therapeutic strategy originating from a challenging weed. This approach holds promise for potential applications in both bioimaging and the combined use of phyto-photothermal therapy for cancer treatment.Keywords: bioactive liposome, hyptis suaveolens, photothermal therapy, lung cancer
Procedia PDF Downloads 971521 Electro-Mechanical Response and Engineering Properties of Piezocomposite with Imperfect Interface
Authors: Rattanan Tippayaphalapholgul, Yasothorn Sapsathiarn
Abstract:
Composites of piezoelectric materials are widely use in practical applications such as nondestructive testing devices, smart adaptive structures and medical devices. A thorough understanding of coupled electro-elastic response and properties of piezocomposite are crucial for the development and design of piezoelectric composite materials used in advanced applications. The micromechanics analysis is employed in this paper to determine the response and engineering properties of the piezocomposite. A mechanical imperfect interface bonding between piezoelectric inclusion and polymer matrix is taken into consideration in the analysis. The micromechanics analysis is based on the Boundary Element Method (BEM) together with the periodic micro-field micromechanics theory. A selected set of numerical results is presented to investigate the influence of volume ratio and interface bonding condition on effective piezocomposite material coefficients and portray basic features of coupled electroelastic response within the domain of piezocomposite unit cell.Keywords: effective engineering properties, electroelastic response, imperfect interface, piezocomposite
Procedia PDF Downloads 234