Search results for: primary cell line establishment

Authors: Shanshan Guo, Xiaoying Zhu, Dominik Jańczewski, Koon Gee Neoh

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Surface charge and wettability are two prominent physical factors governing cell adhesion and have been extensively studied in the literature. However, a comparison between the two driving forces in terms of their independent and cooperative effects in affecting cell adhesion is rarely explored on a systematic and quantitative level. Herein, we formulate a protocol which allows two-dimensional and independent control over both surface charge and wettability. This protocol enables the unambiguous comparison of the effects of these two properties on cell adhesion. This strategy is implemented by controlling both the relative thickness of polyion layers in the layer-by-layer assembly and the polyion side chain chemical structures. The 2D property matrix spans surface isoelectric point ranging from 5 to 9 and water contact angle from 35º to 70º, with other interferential factors (e.g. roughness) eliminated. The interplay between these two surface variables influences 3T3 fibroblast cell adhesion. The results show that both surface charge and wettability have an effect on its adhesion. The combined effects of positive charge and hydrophilicity led to the highest cell adhesion whereas negative charge and hydrophobicity led to the lowest cell adhesion. Our design strategy can potentially form the basis for studying the distinct behaviors of electrostatic force or wettability driven interfacial phenomena and serving as a reference in future studies assessing cell adhesion to surfaces with known charge and wettability within the property range studied here.

Keywords: cell adhesion, layer-by-layer, surface charge, surface wettability

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Authors: Masood Sattari

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Using fuel cell technology to generate electricity for large commercial and industrial sites is a growing segment in the fuel cell industry. The installation of these systems involves design, permitting, procurement of long-lead electrical equipment, and construction involving multiple utilities. The installation of each fuel cell system requires the same amount of coordination as the construction of a new structure requiring a foundation, gas, water, and electricity. Each of these components provide variables that can delay and possibly eliminate a new project. As the manufacturing process and efficiency of fuel cell systems improves, so must the installation methods to prevent a ‘bottle-neck’ in the installation phase of the deployment. Installation methodologies to install the systems vary among companies and this paper will examine the methodologies, describe the benefits and drawbacks for each, and provide guideline for the industry to improve overall installation efficiency.

Keywords: construction, installation, methodology, procurement

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Authors: K. Urukundu, K. A. Aravind, Pradeep M. Nirgude, K. Sandhya

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Transmission of bulk power over a long distance through HVDC transmission lines is gaining importance. This is because the transfer of bulk power through HVDC, from generating stations to load centers over long distances is more economical. However, these HVDC transmission lines create environmental and interference effects under the right of way of the line due to the ionization of the surrounding atmosphere in the vicinity of HVDC lines. The measurement of ground-level electric field and ionic current density is essential for the evaluation of human effects due to electromagnetic interference of the HVDC transmission line. In this paper, experimental laboratory results of the ground-level electric field under the miniature model of 800 kV monopole HVDC line of length 8 meters are presented in lateral configuration with different heights of the conductor from the ground plane. The results are compared with the simulated test results obtained through Finite Element based software.

Keywords: bundle, conductor, hexagonal, transmission line, ground-level electric field

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Authors: M. M. Rahman, A. Liu, A. Frostegard, J. Frostegard

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The human immune system plays an essential role in cardiovascular disease (CVD) and atherosclerosis. Our earlier studies showed that major immunocompetent cells including T cells are activated by phosphorylcholine epitope. Further, we have determined for the first time in a clinical cohort that antibodies against phosphorylcholine (anti-PC) are negatively and independently associated with the development of atherosclerosis and thus a low risk of cardiovascular diseases. It is still unknown whether activated T cells play a role in anti-PC production. Here we aim to clarify the role of T cells in anti-PC production. B cell alone, or with CD3 T, CD4 T or with CD8 T cells were cultured in polystyrene plates to examine anti-PC IgM production. In addition to mixed B cell with CD3 T cell culture, B cells with CD3 T cells were also cultured in transwell co-culture plates. Further, B cells alone and mixed B cell with CD3 T cell cultures with or without anti-HLA 2 antibody were cultured for 6 days. Anti-PC IgM was detected by ELISA in independent experiments. More than 8 fold higher levels of anti-PC IgM were detected by ELISA in mixed B cell with CD3 T cell cultures in comparison to B cells alone. After the co-culture of B and CD3 T cells in transwell plates, there were no increased antibody levels indicating that B and T cells need to interact to augment anti-PC IgM production. Furthermore, anti-PC IgM was abolished by anti-HLA 2 blocking antibody in mixed B and CD3 T cells culture. In addition, the lack of increased anti-PC IgM in mixed B with CD8 T cells culture and the increased levels of anti-PC in mixed B with CD4 T cells culture support the role of helper T cell for the anti-PC IgM production. Atherosclerosis is a major cause of cardiovascular diseases, but anti-PC IgM is a protection marker for atherosclerosis development. Understanding the mechanism involved in the anti-PC IgM regulation could play an important role in strategies to raise anti-PC IgM. Studies suggest that anti-PC is T-cell independent antibody, but our study shows the major role of T cell in anti-PC IgM production. Activation of helper T cells by immunization could be a possible mechanism for raising anti-PC levels.

Keywords: anti-PC, atherosclerosis, aardiovascular diseases, phosphorylcholine

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Authors: Celine Shi Ying Lee, Jong Jian Lee

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Background: Infectious keratitis is one of the leading causes of blindness worldwide. Prompt treatment with effective medication will control the infection early, preventing corneal scarring and visual loss. fluoroquinolones ophthalmic medication is used because of its broad-spectrum properties, potency, good intraocular penetration, and low toxicity. The study aims to evaluate the treatment outcome of corneal ulcers using Levofloxacin 1.5% ophthalmic solution (LVFX) with adjuvant oral ciprofloxacin when indicated and apply this treatment strategy in primary health care as first-line treatment. Methods: Patients with infective corneal ulcer treated in an eye center were recruited. Inclusion criteria includes Corneal infection consistent with bacterial keratitis, single or multiple small corneal ulcers. Treatment regime: LVFX hourly for the first 2 days, 2 hourly from the 3rd day, and 3 hourly on the 5th day of review. Adjuvant oral ciprofloxacin 500mg BD was administered for 5 days if there were multiple corneal ulcers or when the location of the cornea ulcer was central or paracentral. Results: 47 subjects were recruited. There were 16 (34%) males and 31 (66%) females. 40 subjects (85%) were contact lens (CL) related to corneal ulcer, and 7 subjects (15%) were non-contact lens related. 42 subjects (89%) presented with one ulcer, of which 20 of them (48%) needed adjuvant therapy. 5 subjects presented with 2 or 3 ulcers, of which 3 needed adjuvant therapy. A total of 23 subjects (49%) was given adjuvant therapy (oral ciprofloxacin 500mg BD for 5 days).21 of them (91%) were CL related. All subjects recovered fully, and the average duration of treatment was 3.7 days, with 49% of the subjects resolved on the 3rd day, 38% on the 5thday of and 13% on the 7thday. All subjects showed symptoms of relief of pain, light-sensitivity, and redness on the 3rd day with full visual recovery post-treatment. No adverse drug reactions were recorded. Conclusion: Our treatment regime demonstrated good clinical outcome as first-line treatment for corneal ulcers. A corneal ulcer is a common eye condition in Singapore, mainly due to CL wear. Pseudomonas aeruginosa is the most frequent and potentially sight-threatening pathogen involved in CL related corneal ulcer. Coagulase-negative Staphylococci, Staphylococcus aureus, and Streptococcus Pneumoniae were seen in non-CL users. All these bacteria exhibit good sensitivity rates to ciprofloxacin and levofloxacin. It is therefore logical in our study to use LVFX Eyedrops and adjuvant ciprofloxacin oral antibiotics when indicated as first line treatment for most corneal ulcers. Our study of patients, both CL related and non-CL related, have shown good clinical response and full recovery using the above treatment strategy. There was also a full restoration of visual acuity in all the patients. Eye-trained primary Healthcare practitioners can consider adopting this treatment strategy as first line treatment in patients with corneal ulcers. This is relevant during the COVID pandemic, where hospitals are overwhelmed with patients and in regions with limited access to specialist eye care. This strategy would enable early treatment with better clinical outcome.

Keywords: corneal ulcer, levofloxacin hydrate, treatment strategy, ciprofloxacin

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Authors: Chijioke Okafor, Malik Maaza, Touhami Mokrani

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Proton exchange membrane, PEM was developed and tested for potential application in fuel cell. Nafion was electrospun to nanofiber network with the aid of poly(ethylene oxide), PEO, as a carrier polymer. The matrix polymer was crosslinked with Norland Optical Adhesive 63 under UV after compacting and annealing. The welded nanofiber mat was characterized for morphology, proton conductivity, and methanol permeability, then tested in a single cell test station. The results of the fabricated nanofiber membrane showed a proton conductivity of 0.1 S/cm at 25 oC and higher fiber volume fraction; methanol permeability of 3.6x10^-6 cm2/s and power density of 96.1 and 81.2 mW/cm2 for 5M and 1M methanol concentration respectively.

Keywords: fuel cell, nafion, nanofiber, permeability

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Authors: G. Cunningham, E. Cantor, X. Wu, F. Shen, G. Jiang, S. Philips, C. Bales, Y. Xiao, T. R. Cummins, J. C. Fehrenbacher, B. P. Schneider

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Background: Taxane-induced peripheral neuropathy (TIPN) is the most devastating survivorship issue for patients receiving therapy. Dose reductions due to TIPN in the curative setting lead to inferior outcomes for African American patients, as prior research has shown that this group is more susceptible to developing severe neuropathy. The mechanistic underpinnings of TIPN, however, have not been entirely elucidated. While it would be appealing to use primary tissue to study the development of TIPN, procuring nerves from patients is not realistically feasible, as nerve biopsies are painful and may result in permanent damage. Therefore, our laboratory has investigated paclitaxel-induced neuronal morphological and molecular changes using an ex vivo model of human-induced pluripotent stem cell (iPSC)-derived neurons. Methods: iPSCs are undifferentiated and endlessly dividing cells that can be generated from a patient’s somatic cells, such as peripheral blood mononuclear cells (PBMCs). We successfully reprogrammed PBMCs into iPSCs using the Erythroid Progenitor Reprograming Kit (STEMCell Technologiesᵀᴹ); pluripotency was verified by flow cytometry analysis. iPSCs were then induced into neurons using a differentiation protocol that bypasses the neural progenitor stage and uses selected small-molecule modulators of key signaling pathways (SMAD, Notch, FGFR1 inhibition, and Wnt activation). Results: Flow cytometry analysis revealed expression of core pluripotency transcription factors Nanog, Oct3/4 and Sox2 in iPSCs overlaps with commercially purchased pluripotent cell line UCSD064i-20-2. Trilineage differentiation of iPSCs was confirmed with immunofluorescent imaging with germ-layer-specific markers; Sox17 and ExoA2 for ectoderm, Nestin, and Pax6 for mesoderm, and Ncam and Brachyury for endoderm. Sensory neuron markers, β-III tubulin, and Peripherin were applied to stain the cells for the maturity of iPSC-derived neurons. Patch-clamp electrophysiology and calcitonin gene-related peptide (CGRP) release data supported the functionality of the induced neurons and provided insight into the timing for which downstream assays could be performed (week 4 post-induction). We have also performed a cell viability assay and fluorescence-activated cell sorting (FACS) using four cell-surface markers (CD184, CD44, CD15, and CD24) to select a neuronal population. At least 70% of the cells were viable in the isolated neuron population. Conclusion: We have found that these iPSC-derived neurons recapitulate mature neuronal phenotypes and demonstrate functionality. Thus, this represents a patient-derived ex vivo neuronal model to investigate the molecular mechanisms of clinical TIPN.

Keywords: chemotherapy, iPSC-derived neurons, peripheral neuropathy, taxane, paclitaxel

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Authors: Djaafar F., Hadri B., Bachir G.

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We studied mainly the influence of temperature, thickness, molar fraction and the doping of the various layers (emitter, base, BSF and window) on the performances of a photovoltaic solar cell. In a first stage, we optimized the performances of the InGaP/GaAs dual-junction solar cell while varying its operation temperature from 275°K to 375 °K with an increment of 25°C using a virtual wafer fabrication TCAD Silvaco. The optimization at 300°K led to the following result Icc =14.22 mA/cm2, Voc =2.42V, FF =91.32 %, η = 22.76 % which is close with those found in the literature. In a second stage ,we have varied the molar fraction of different layers as well their thickness and the doping of both emitters and bases and we have registered the result of each variation until obtaining an optimal efficiency of the proposed solar cell at 300°K which was of Icc=14.35mA/cm2,Voc=2.47V,FF=91.34,and η =23.33% for In(1-x)Ga(x)P molar fraction( x=0.5).The elimination of a layer BSF on the back face of our cell, enabled us to make a remarkable improvement of the short-circuit current (Icc=14.70 mA/cm2) and a decrease in open circuit voltage Voc and output η which reached 1.46V and 11.97% respectively. Therefore, we could determine the critical parameters of the cell and optimize its various technological parameters to obtain the best performance for a dual junction solar cell. This work opens the way with new prospects in the field of the photovoltaic one. Such structures will thus simplify the manufacturing processes of the cells; will thus reduce the costs while producing high outputs of photovoltaic conversion.

Keywords: modeling, simulation, multijunction, optimization, silvaco ATLAS

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Authors: Kristel Dame Bañez Sumagaysay, Marie Victoria Cruz-javier

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The current subject is a case of a 21 year-old woman at 29 1/7 weeks of gestation with Pre-B cell Acute Lymphoblastic Leukemia who was admitted to the coronary care unit (CCU) of the St. Luke’s Medical Center-Global City for Severe Acute Respiratory Distress Syndrome (ARDS) secondary to hospital-acquired pneumonia secondary to pneumocystis jiroveci; central line-associated bloodstream infection (E. aerogenes). She presented with chronic hypoxemia caused by Pulmonary edema, probably secondary to heart failure secondary to cardiomyopathy chemotherapy-induced. Due to worsening feto-maternal status, extracorporeal membrane oxygenation (ECMO) for respiratory support was instituted, and an elective cesarean section was done due to multiple maternal factors and deteriorating health status under total intravenous anesthesia assisted by veno-venous extracorporeal membrane oxygenation. She delivered a live preterm newborn male, APGAR Score: 1, 0, 0, birth weight 985 grams, birth length: 40.5cm, small for gestational age.

Keywords: extracorporeal membrane oxygenation, pre-b cell acute lymphoblastic leukemia, severe acute respiratory distress syndrome, ethical dilemmas

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Authors: Samira Khalaji, , Fenneke Klein Jan, Kay-E. Gottschalk, Eugenia Makrantonaki, Karin Scharffetter-Kochanek

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Biological aging is a multi-dimensional process that takes place over a whole range of scales from the nanoscopic alterations within individual cells, over transformations in tissues and organs and to changes of the whole organism. On the single cell level, aging involves mutation of genes, differences in gene expression levels as well as altered posttranslational modifications of proteins. A variety of proteins is affected, including proteins of the cell cytoskeleton and migration machinery. Previous work quantified the expression of cytoskeleton proteins on the gene and protein levels in senescent and young fibroblasts. Their results show that senescent skin fibroblasts have an upregulated expression of the intermediate filament (IF) protein vimentin in contrast to actin and tubulin, which are downregulated. IFs play an important role in providing mechanical stability of cells. However, the mechanical properties of IFs depending on cellular senescence or age of the donor has not been studied so far. Hence, we employed passive microrheology on primary human dermal fibroblasts from female donors with age of 28 years (young) and 86 years (old) as model of in vivo aging and human normal dermal fibroblast from 11-year old male with CPD 17-35 (young) and CPD 58-59 (senescence) as a model of in vitro replicative senescence. In contrast to the expectations, our primary results show no significant differences in the viscoelastic properties of fibroblasts depending on age of the donor or cellular replicative senescence.

Keywords: aging, cytoskeleton, fibroblast, mechanical properties

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Authors: Atefeh Esmailnejad, Gholamreza Nikbakht Brujeni

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Major histocompatibility complex (MHC) is one of the best characterized genetic regions associated with immune responses and controlling disease resistance in chicken. Association of the MHC with a wide range of immune responses makes it a valuable predictive factor for the disease pathogenesis and outcome. In this study, the association of MHC with cell-mediated immune responses was analyzed in commercial broiler chicken. The tandem repeat LEI0258 was applied to investigate the MHC polymorphism. Cell-mediated immune response was evaluated by peripheral blood lymphocyte proliferation assay using MTT method. Association study revealed a significant influence of MHC alleles on cellular immune responses in this population. Alleles 385 and 448 bp were associated with elevated cell-mediated immunity. Haplotypes associated with improved immune responses could be considered as candidate markers for disease resistance and applied to breeding strategies.

Keywords: MHC, cell-mediated immunity, broiler, chicken

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Authors: Yuping Liu, Li Tao, Yin Lu

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Accumulating evidence has been shown that diallyl trisulfide (DATS) from garlic may reduce the risk of developing several types of cancer. In view of the dynamic crosstalk interplayed by tumor cells and platelets in hematogenous metastasis, we demonstrate the effectiveness of DATS on the metastatic behaviors of MDA-MB-435s human breast cancer cell line co-incubated with activated platelets. Indeed, our data identified that DATS significantly blocked platelets fouction induced by PAF, followed by the decreased production of TXB2. DATS was found to dose-dependently suppressed MDA-MB-435s cell migration and invasion in presence of activated platelets by PAF in vitro. Furthermore, the expression, secretion and enzymatic activity of matrix metalloproteinase (MMP)-2/9, as well as the luciferase activity of upstream regulator NF-κB in MDA-MB-435s, were obviously diminished by DATS. In parallel, DATS blocked upstream NF-κB activation signaling complexes composed of extracellular signal-related kinase (ERK) as assessed by measuring the levels of the phosphorylated forms.

Keywords: DATS, ERK, metastasis, MMPs, NF-κB, platelet

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Authors: Maedeh Rahimnejad, Bahman Vahidi, Bahman Ebrahimi Hoseinzadeh, Fatemeh Yazdian, Puria Motamed Fath, Roghieh Jamjah

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Introduction: The intensive labor and high cost of developing new vehicles for controlled drug delivery highlights the need for a change in their discovery process. Computational models can be used to accelerate experimental steps and control the high cost of experiments. Methods: In this work, to better understand the interaction of anti-cancer drug and the nanocarrier with the cell membrane, we have done molecular dynamics simulation using NAMD. We have chosen paclitaxel for the drug molecule and dipalmitoylphosphatidylcholine (DPPC) as a natural phospholipid nanocarrier. Results: Next, center of mass (COM) between molecules and the van der Waals interaction energy close to the cell membrane has been analyzed. Furthermore, the simulation results of the paclitaxel interaction with the cell membrane and the interaction of DPPC as a nanocarrier loaded by the drug with the cell membrane have been compared. Discussion: Analysis by molecular dynamics (MD) showed that not only the energy between the nanocarrier and the cell membrane is low, but also the center of mass amount decreases in the nanocarrier and the cell membrane system during the interaction; therefore they show significantly better interaction in comparison to the individual drug with the cell membrane.

Keywords: anti-cancer drug, center of mass, interaction energy, molecular dynamics simulation, nanocarrier

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Authors: Marcio S. da Silva, Jose Mauricio de B. Bezerra, Antonio E. de A. Nogueira

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This paper aims to make a performance analysis of shield wire transmission lines against atmospheric discharges when it is made the option of sectioning the shield wire and verify if the tolerability of the change. As a goal of this work, it was established to make complete modeling of a transmission line in the ATPDraw program with shield wire grounded in all the towers and in some towers. The methodology used to make the proposed evaluation was to choose an actual transmission line that served as a case study. From the choice of transmission line and verification of all its topology and materials, complete modeling of the line using the ATPDraw software was performed. Then several atmospheric discharges were simulated by striking the grounded shield wires in each tower. These simulations served to identify the behavior of the existing line against atmospheric discharges. After this first analysis, the same line was reconsidered with shield wire segmentation. The shielding wire segmentation technique aims to reduce induced losses in shield wires and is adopted in some transmission lines in Brazil. With the same conditions of atmospheric discharge the transmission line, this time with shield wire segmentation was again evaluated. The results obtained showed that it is possible to obtain similar performances against atmospheric discharges between a shield wired line in multiple towers and the same line with shield wire segmentation if some precautions are adopted as verification of the ground resistance of the wire segmented shield, adequacy of the maximum length of the segmented gap, evaluation of the separation length of the electrodes of the insulator spark, among others. As a conclusion, it is verified that since the correct assessment and adopted the correct criteria of adjustment a transmission line with shielded wire segmentation can perform very similar to the traditional use with multiple earths. This solution contributes in a very important way to the reduction of energy losses in transmission lines.

Keywords: atmospheric discharges, ATPDraw, shield wire, transmission lines

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Authors: Yu-Chen Hu

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Gene editing by CRISPR and gene regulation by microRNA or CRISPR activation have dramatically changed the way to manipulate cellular gene expression and cell fate. In recent years, various gene editing and gene manipulation technologies have been applied to control stem cell differentiation to enhance tissue regeneration. This research will focus on how to develop CRISPR, CRISPR activation (CRISPRa), CRISPR inhibition (CRISPRi), as well as bi-directional CRISPR-AI gene regulation technologies to control cell differentiation and bone regeneration. Moreover, in this study, CRISPR/Cas13d-mediated RNA editng for miRNA editing and bone regeneration will be discussed.

Keywords: gene therapy, bone regeneration, stem cell, CRISPR, gene regulation

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Authors: Safiullah Abdullahi

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This paper describes the design of a digital control system which controls the speed and direction of a robot. The robot is expected to follow a black thick line with the highest possible speed and lowest error around the line. The control system of the robot will correct for the angle error that is made between the frame axis of the robot and the line. The cause for error is the difference in speed of the two driving wheels of the robot which are driven by two separate DC motors, whereas the speed difference in wheels is due to the un-modeled fraction that is available in the wheels with different magnitudes in each. The control scheme is that a number of photo sensors are mounted in the front of the robot and report their position in reference to the black line to the digital controller. The controller then, evaluates the position error and generates the needed duty cycle for the related wheel motor to drive it faster or slower.

Keywords: digital control, robot, controller, control system

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Authors: Fatemeh Abbasi, Arne Hofemeier, Timo Betz

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Muscle growth and regeneration critically depend on satellite cells (SCs) which are muscle stem cells located between the basal lamina and myofibres. Upon damage, SCs become activated, enter the cell cycle, and give rise to myoblasts that form new myofibres, while a sub-population self-renew and re-populate the muscle stem cell niche. In aged muscle as well as in certain muscle diseases such as muscular dystrophy, some of the SCs lose their regenerative ability. Although it is demonstrated that the chemical composition of SCs quiescent niche is different from the activated niche, the mechanism initially activated in the SCs remains unknown. While extensive research efforts focused on potential chemical activation, no such factor has been identified to the author’s best knowledge. However, it is substantiated that niche mechanics affects SCs behaviors, such as stemness and engraftment. We hypothesize that mechanical stress in the healthy niche (homeostasis) is different from the regenerative niche and that this difference could serve as an early signal activating SCs upon fiber damage. To investigate this hypothesis, we develop a biomimetic system to reconstitute both, the mechanical and the chemical environment of the SC niche. Cells will be confined between two elastic polyacrylamide (PAA) hydrogels with controlled elastic moduli and functionalized surface chemistry. By controlling the distance between the PAA hydrogel surfaces, we vary the compression forces exerted by the substrates on the cells, while the lateral displacement of the upper hydrogel will create controlled shear forces. To establish such a system, a simplified system is presented. We engineered a sandwich-like configuration of two elastic PAA layer with stiffnesses between 1 and 10 kPa and confined a precursor myoblast cell line (C2C12) in between these layers. Our initial observations in this sandwich model indicate that C2C12 cells show different behaviors under mechanical compression if compared to a control one-layer gel without compression. Interestingly, this behavior is stiffness-dependent. While the shape of C2C12 cells in the sandwich consisting of two stiff (10 kPa) layers was much more elongated, showing almost a neuronal phenotype, the cell shape in a sandwich situation consisting of one stiff and one soft (1 kPa) layer was more spherical. Surprisingly, even in proliferation medium and at very low cell density, the sandwich situation stimulated cell differentiation with increased striation and myofibre formation. Such behavior is commonly found for confluent cells in differentiation medium. These results suggest that mechanical changes in stiffness and applied pressure might be a relevant stimulation for changes in muscle cell behavior.

Keywords: C2C12 cells, compression, force, satellite cells, skeletal muscle

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Authors: Furqan M. Kadhum, Asmaa A. Hussein, Maysaa Ch. Hatem

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Hyaluronidase enzyme was purified from the clinical isolate Staphyloccus aureus in three purification steps, first by precipitation with 90% saturated ammonium sulfate, ion exchange chromatography on DEAE-Cellulose, and gel filtration chromatography throughout Sephacryl S-300. Specific activity of the purified enzyme was reached 930 U/mg protein with 7.4 folds of purification and 46.5% recovery. The enzyme has an average molecular weight of about 69 kDa, with an optimum pH of enzyme activity and stability at pH 7, also the optimum temperature for activity was 37oC. The enzyme was stable with full activity at a temperature ranged between 30-40 oC. Metal ions showed variable inhibitory degree with the strongest effect for Fe+3, however, the chelating and reducing agents had no or little effects. Cytotoxic studies for purified and crude hyaluronidase against cancer cell Hep G2 type at different enzyme concentrations and exposure times showed that the inhibition effect of both crude and purified enzyme increased by increasing the enzyme concentration with no change was observed at 24hr, while at 48 and 72 hrs the same inhibition rate were observed for purified enzyme and differ for the crude filtrate.

Keywords: hyaluronidase, S. aureus, metal ions, cytotoxicity

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Authors: J. L. Liu, L. Wang, B. Zhu, J. Zhou, W. D. Gao

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Fabric textures are very common in our daily life. However, we never explore the representation of fabric textures from neuroscience view. Theoretical studies suggest that primary visual cortex (V1) uses a sparse code to efficiently represent natural images. However, how the simple cells in V1 encode the artificial textures is still a mystery. So, here we will take fabric texture as stimulus to study the response of independent component analysis that is established to model the receptive field of simple cells in V1. Experimental results based on 140 classical fabric images indicate that the receptive fields of simple cells have obvious selectivity in orientation, frequency, and phase when drifting gratings are used to determine their tuning properties. Additionally, the distribution of optimal orientation and frequency shows that the patch size selected from each original fabric image has a significant effect on the frequency selectivity.

Keywords: fabric texture, receptive filed, simple cell, spare coding

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Authors: Elena Petersen, Inna Kornienko, Svetlana Guryeva, Sergey Dobdin, Anatoly Skripal, Andrey Usanov, Dmitry Usanov

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One of the most important unsolved problems in modern medicine is the increase of chronic diseases that lead to organ dysfunction or even complete loss of function. Current methods of treatment do not result in decreased mortality and disability statistics. Currently, the best treatment for many patients is still transplantation of organs and/or tissues. Therefore, finding a way of correct artificial matrix biofabrication in case of limited number of natural organs for transplantation is a critical task. One important problem that needs to be solved is development of a nondestructive and noninvasive method to analyze dynamic changes of mechanical characteristics of a matrix with minimal side effects on the growing cells. This research was focused on investigating the properties of matrix as a marker of graft condition. In this study, the collagen gel with human primary dermal fibroblasts in suspension (60, 120, 240*103 cells/mL) and collagen gel with cell spheroids were used as model objects. The stiffness and elasticity characteristics were evaluated by a semiconductor laser autodyne. The time and cell concentration dependency of the stiffness and elasticity were investigated. It was shown that these properties changed in a non-linear manner with respect to cell concentration. The maximum matrix stiffness was observed in the collagen gel with the cell concentration of 120*103 cells/mL. This study proved the opportunity to use the mechanical properties of matrix as a marker of graft condition, which can be measured by noninvasive semiconductor laser autodyne technique.

Keywords: graft, matrix, noninvasive method, regenerative medicine, semiconductor laser autodyne

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Authors: Mohammad Ansari, Ahmed Ismail, Satinder Mann

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Overcrowding in Emergency departments (ED) of United Kingdom has become a common problem. Urgent Care centres were developed nearly a decade ago to reduce pressure on EDs. Unfortunately, the development of Urgent Care centres has failed to produce the projected effects. It was thought that nearly 40% patients attending ED would go to Urgent Care centres and these would be staffed by Primary care Physicians. Data reveals that no more than 20% patients were seen by Primary Care Physicians even when the Urgent Care Centre was based in the ED. This study was carried out at the ED of George Eliot Hospital, Nuneaton, UK where the Urgent Care centre was based in the ED and employed Primary Care Physicians with special interest in trauma for nearly one year. This was then followed by a Primary Care Physician and Advanced Nurse Practitioner. We compared the number of patients seen during these periods and the cost-effectiveness of the service.We randomly selected a week of patients seen by Primary Care Physicians with special interest in Trauma and by Primary Care Physicians and the Advanced Nurse Practitioner. We compared the number and type of patients seen during these two periods. Nearly 38% patients were seen by Primary care Physician with special interest in Trauma, whilst only 14.3% patients were seen by the Primary care Physician and Advanced Nurse Practitioner. The Primary Care Physicians with special interest in trauma were paid less. Our study confirmed that unless Primary Care Physicians are able to treat minor trauma and interpret x-rays, the urgent care service is not going to be cost effective. Numerous previous studies have shown that 15 to 20% patients attending ED can be treated by Primary Care Physicians who do not require any investigations for their management. It is advantageous to have Urgent Care Centres within the ED because if the patient deteriorates they can be transferred to ED. We recommend that the Urgent care Centres should be a part of ED. Our study shows that Urgent care Centres in the ED can be helpful and cost effective if staffed by either senior Emergency Physicians or Primary Care Physicians with special interest and experience in the management of minor trauma.

Keywords: urgent care centres, primary care physician, advanced nurse practitioner, trauma

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Authors: S. AliReza Mirghasemi, Zameer Hussain, Mohammad Saleh Sadeghi, Narges Rahimi Gabaran, Mohamadreza Baghaban Eslaminejad

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Background: Despite promising results have shown by osteogenic cell-based demineralized bone matrix composites, they need to be optimized for grafts that act as structural frameworks in load-bearing defects. The purpose of this experiment is to determine the effect of bone-marrow-mesenchymal-stem-cells seeding on partially demineralized laser-perforated structural allografts that have been implanted in critical femoral defects. Materials and Methods: P3 stem cells were used for graft seeding. Laser perforation in four rows of three holes was achieved. Cell-seeded grafts were incubated for one hour until they were planted into the defect. We used four types of grafts: partially demineralized only (Donly), partially demineralized stem cell seeded (DST), partially demineralized laser-perforated (DLP), and partially demineralized laser-perforated stem cell seeded (DLPST). histologic and histomorphometric analysis were performed at 12 weeks. Results: Partially demineralized laser-perforated had the highest woven bone formation within graft limits, stem cell seeded demineralized laser-perforated remained intact, and the difference between partially demineralized only and partially demineralized stem cell seeded was insignificant. At interface, partially demineralized laser-perforated and partially demineralized only had comparable osteogenesis, but partially demineralized stem cell seeded was inferior. The interface in stem cell seeded demineralized laser-perforated was almost replaced by distinct endochondral osteogenesis with higher angiogenesis in the vicinity. Partially demineralized stem cell seeded and stem cell seeded demineralized laser-perforated graft surfaces had extra vessel-ingrowth-like porosities, a sign of delayed resorption. Conclusion: This demonstrates that simple cell-based composites are not optimal and necessitates the supplementation of synergistic stipulations and surface changes.

Keywords: structural bone allograft, partial demineralization, laser perforation, mesenchymal stem cell

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Authors: Winnie Wing Mui So, Yu Chen

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This study investigated primary school teachers’ and students’ perceptions of science learning in an e-learning environment. This study used a multiple case study design and involved eight science teachers and their students from four Hong Kong primary schools. The science topics taught included ‘season and weather’ ‘force and movement’, ‘solar and lunar eclipse’ and ‘living things and habitats’. Data were collected through lesson observations, interviews with teachers, and interviews with students. Results revealed some differences between the teachers’ and the students’ perceptions regarding the usefulness of e-learning resources, the organization of student-centred activities, and the impact on engagement and interactions in lessons. The findings have implications for the more effective creation of e-learning environments for science teaching and learning in primary schools.

Keywords: e-learning, science education, teacher' and students' perceptions, primary schools

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Authors: Reshu Saxena, R. K. Tripathi

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HIV-1 transmission and spread involves significant host-virus interaction. Potential targets for prevention of HIV-1 lies at the site of mucosal barriers. Thus a better understanding of how HIV-1 infects target cells at such sites and lead their invasion is required, with prime focus on the host determinants regulating HIV-1 spread. HIV-1 Nef is important for viral infectivity and pathogenicity. It promotes HIV-1 replication, facilitating immune evasion by interacting with various host factors and altering cellular pathways via multiple protein-protein interactions. In this study nef was sequenced from HIV-1 patients, and showed specific mutations revealing sequence variability in nef. To explore the difference in Nef functionality based on sequence variability we have studied the effects of HIV-1 Nef in human SupT1 T cell line and (THP-1) monocyte-macrophage cell lines through proteomics approach. 2D-Gel Electrophoresis in control and Nef-transfected SupT1 cells demonstrated several differentially expressed proteins with significant modulation of alpha-enolase. Through further studies, effects of Nef on alpha-enolase regulation were found to be cell lineage-specific, being stimulatory in macrophages/monocytes, inhibitory in T cells and without effect in HEK-293 cells. Cell migration and invasion studies were employed to determine biological function affected by Nef mediated regulation of alpha-enolase. Cell invasion was enhanced in THP-1 cells but was inhibited in SupT1 cells by wildtype nef. In addition, the modulation of enolase and cell invasion remained unaffected by a unique nef variant. These results indicated that regulation of alpha-enolase expression and invasive property of host cells by Nef is sequence specific, suggesting involvement of a particular motif of Nef. To precisely determine this site, we designed a heptapeptide including the suggested alpha-enolase regulating sequence of nef and a nef mutant with deletion of this site. Macrophages/monocytes being the major cells affected by HIV-1 at mucosal barriers, were particularly investigated by the nef mutant and peptide. Both the nef mutant and heptapeptide led to inhibition of enhanced enolase expression and increased invasiveness in THP-1 cells. Together, these findings suggest a possible mechanism of host invasion by HIV-1 through Nef mediated regulation of alpha-enolase and identifies a potential therapeutic target for HIV-1 entry at mucosal barriers.

Keywords: HIV-1 Nef, nef variants, host-virus interaction, tissue invasion

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Authors: Adel M. Mahmoud

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This Investigation was conducted to determine the productivity and water use efficiency for new sunflower genotypes. Ten sunflower genotypes were evaluated under drip irrigation using two treatments of. Results indicate that decreasing the amount of irrigation water from 1500 to 1130 mm/hectar significantly reduced all studied traits. Mutation (M1-63) surpassed all the other one genotypes in seed yield and WUE. Lines which gave the highest yield of the seed have water use efficiency under drought conditions higher than water use efficiency under normal irrigation. The lowest depression in seed yield due to drought conditions has been registered for Line 20, Line M1-63 and Sakha 53 genotypes (11 , 18 and 16 %, respectively). Genotypes (Line 20 , Line M1-63 and Sakha 53) are more tolerant to drought than others and we can used its in breeding program to develop sunflower hybrids suitable for cultivation under drought condition.

Keywords: sunflower genotypes, water use efficiency, mutation, inbred lines

Procedia PDF Downloads 361

Authors: Joanna Kochanska, Dagmara Gornicka, Anna Burduk

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The article examines the process of implementing a new assembly line in a manufacturing enterprise of the household appliances industry area. At the initial stages of the project, a decision was made that one of its foundations should be the concept of lean management. Because of that, eliminating as many errors as possible in the first phases of its functioning was emphasized. During the start-up of the line, there were identified and documented all production losses (from serious machine failures, through any unplanned downtime, to micro-stops and quality defects). During 6 weeks (line start-up period), all errors resulting from problems in various areas were analyzed. These areas were, among the others, production, logistics, quality, and organization. The aim of the work was to analyze the occurrence of production failures during the initial phase of starting up the line and to propose a method for determining their critical level during its full functionality. There was examined the repeatability of the production losses in various areas and at different levels at such an early stage of implementation, by using the methods of statistical process control. Based on the Pareto analysis, there were identified the weakest points in order to focus improvement actions on them. The next step was to examine the effectiveness of the actions undertaken to reduce the level of recorded losses. Based on the obtained results, there was proposed a method for determining the critical failures level in the studied areas. The developed coefficient can be used as an alarm in case of imbalance of the production, which is caused by the increased failures level in production and production support processes in the period of the standardized functioning of the line.

Keywords: production failures, level of production losses, new production line implementation, assembly line, statistical process control

Procedia PDF Downloads 114

Authors: Melliti Rihab, Zaeid Sonia, Khechine Wiem, Daldoul Amira

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Introduction: Lung cancer is the leading cause of cancer death. Its incidence is increasing, and its prognosis remains pejorative. We present the clinical, pathological, and therapeutic characteristics of bronchopulmonary cancer (BPC) in Tunisia. Methods: Retrospective study including patients followed in the oncology department of the University Hospital of Monastir between April 2014 and December 2021 suffering from lung cancer. Results: These are 117 patients, including 86.3% men and 13.7% women (sex ratio 6.3). The average age was 64 years ± 9 (37-83), with 95.7% being over 50 years old. Patients were smokers in 82% of cases. The clinical signs were dominated by chest pain (27.5%) and dyspnea in 21.1% of cases. In 6 patients, an episode of COVID-19 infection revealed the diagnosis. Half of the patients had a PS between 0 and 1. Small cell lung cancer was present in 18 patients (15.4%). The majority of non small cell lung cancer was of the adenocarcinoma type (68.7%). The diagnosis was late (stage IV) in 62.4% of cases. BPC was metastatic to bone (52%), contralateral lung (25.9%), and brain (27.3%). Patients were oligometastatic in 26% of cases. Surgery and radiotherapy were performed respectively in 14.5% and 23.1% of cases. Three-quarters of the patients had had nutrition (75.2%). The ROS1 mutation was present in 1 patient. PDL-1 expression was >40% in 2 patients. Survival was mean eight months ± 7.4. Conclusion: Lung cancer is diagnosed at a late stage in Tunisia. The lack of molecular study for non-small cell PBC and the lack of marketing authorization for tyrosine kinase inhibitors in Tunisia make the management incomplete.

Keywords: SCLC, NCSLC, ROS1, PDL1

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Authors: Stephen Adeniyi Adefegha, Vitor Mostardeiro, Vera Maria Morsch, Ademir F. Morel, Ivana Beatrice Manica Da Cruz, Sabrina Somacal Maria Rosa Chitolina Schetinger

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Moringa stenopetala is often consumed as food and used in folkloric medicine for the management of several diseases. Purpose: This study was set up in order to assess the effect of aqueous extract of Moringa stenopetala on cell viability and oxidative stress biomarkers in BV-2 microglial cells. Aqueous extracts of M. stenopetala were prepared, lyophilized and reconstituted in 0.5% dimethylsulphoxide (DMSO). Cells were treated with M. stenopetala extracts (0.1 - 100 µg/ml) for cell viability and nitric oxide (NO) production tests. However, M. stenopetala extract (50 µg/ml) was used in the treatment of cells for the determination of protein carbonyl content and reactive oxygen species (ROS) level. Incubation of BV-2 microglia cell with M. stenopetala extract maintained cell viability, diminished NO and ROS levels, and reduced protein carbonyl contents Chlorogenic acid, rutin, kaempferol and quercetin derivatives were the main phenolic compounds identified in M. stenopetala leaf extract. These phenolic compounds present in M. stenopetala may be responsible for the mitigation of oxidative stress in BV-2 microglial cells.

Keywords: oxidative stress, BV-2 microglial cell, Moringa stenopetala, cell viability, antioxidant

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Authors: Robert P. Dufresne, Hamid Arabzadeh

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The current study investigates the effectiveness of a new form of permanent baseboard insulators with an umbrella action, hereinafter referred to as Coronary Insulator, in supporting and protecting the assembly of electrodes immersed in an electrolytic cell and in increasing the lifespan of the lateral sides of the electrolytic cell, in both electro-winning and electro-refinery method. The advantages of using a coronary insulator in addition to the top capping board (equipotential insulator) were studied compared to the conventional assembly of an electrolytic cell. Then, a thermal imaging technique was utilized during high-temperature thermal (heat transfer) tests for sample cell walls with and without coronary insulators in their assembly to show the effectiveness of coronary insulators in protecting the cell wall under extreme conditions. It was shown that, unlike the conventional assembly, which is highly prone to damages to the cell wall under thermal shocks, the presence of coronary insulator can significantly increase the level of protection of the cell due to their ultra-high thermal and chemical resistance, as well as decreasing the replacement frequency of insulators to almost zero. Besides, the results of the study showed that the test assembly with the coronary insulator provides better consistency in positioning and, subsequently, better contact, compared to the conventional method, which reduces the chance of electric short-circuit in the system.

Keywords: capping board, coronary insulator, electrolytic cell, thermal shock.

Procedia PDF Downloads 174

Authors: Iulianna Taritsa, Kuldeep Neote, Eric Fossel

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Lysosome-induced immunogenic cell death (LIICD) is a powerful mechanism of targeting cancer cells that kills circulating malignant cells and primes the host’s immune cells against future remission. Current immunotherapies for cancer are limited in preventing recurrence – a gap that can be bridged by training the immune system to recognize cancer neoantigens. Lysosomal leakage can be induced therapeutically to traffic antigens from dying cells to dendritic cells, which can later present those tumorigenic antigens to T cells. Previous research has shown that oxidative agents administered in the tumor microenvironment can initiate LIICD. We generated a fusion protein between an oxidative agent known as xanthine oxidase (XO) and a mini-antibody specific for EGFR/HER2-sensitive breast tumor cells. The anti-EGFR single domain antibody fragment is uniquely sourced from llama, which is functional without the presence of a light chain. These llama micro-antibodies have been shown to be better able to penetrate tissues and have improved physicochemical stability as compared to traditional monoclonal antibodies. We demonstrate that the fusion protein created is stable and can induce early markers of immunogenic cell death in an in vitro human breast cancer cell line (SkBr3). Specifically, we measured overall cell death, as well as surface-expressed calreticulin, extracellular ATP release, and HMGB1 production. These markers are consensus indicators of ICD. Flow cytometry, luminescence assays, and ELISA were used respectively to quantify biomarker levels between treated versus untreated cells. We also included a positive control group of SkBr3 cells dosed with doxorubicin (a known inducer of LIICD) and a negative control dosed with cisplatin (a known inducer of cell death, but not of the immunogenic variety). We looked at each marker at various time points after cancer cells were treated with the XO/antibody fusion protein, doxorubicin, and cisplatin. Upregulated biomarkers after treatment with the fusion protein indicate an immunogenic response. We thus show the potential for this fusion protein to induce an anticancer effect paired with an adaptive immune response against EGFR/HER2+ cells. Our research in human cell lines here provides evidence for the success of the same therapeutic method for patients and serves as the gateway to developing a new treatment approach against breast cancer.

Keywords: apoptosis, breast cancer, immunogenic cell death, lysosome

Procedia PDF Downloads 187