Search results for: human cell line
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 13035

Search results for: human cell line

12645 Transition to Hydrogen Cities in Korea and Japan

Authors: Minhee Son, Kyung Nam Kim

Abstract:

This study explores the plan of the Korean and Japanese governments to transition into the hydrogen economy. Two motor companies, Hyundai Motor Company from Korea and Toyota from Japan, released the Hydrogen Fuel Cell Vehicle to monopolize the green energy automobile market. Although, they are the main countries which emit greenhouse gas, hydrogen energy can bring from a certain industry places, such as chemical plants and steel mills. Recent, the two countries have been focusing on the hydrogen industry including a fuel cell vehicle, a hydrogen station, a fuel cell plant, a residential fuel cell. The purpose of this paper is to find out the differences of the policies in the two countries to be hydrogen societies. We analyze the behavior of the public and private sectors in Korea and Japan about hydrogen energy and fuel cells for the transition of the hydrogen economy. Finally we show the similarities and differences of both countries in hydrogen fuel cells. And some cities have feature such as Hydrogen cities. Hydrogen energy can make impact environmental sustainability.

Keywords: fuel cell, hydrogen city, hydrogen fuel cell vehicle, hydrogen station, hydrogen energy

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12644 CD97 and Its Role in Glioblastoma Stem Cell Self-Renewal

Authors: Niklas Ravn-Boess, Nainita Bhowmick, Takamitsu Hattori, Shohei Koide, Christopher Park, Dimitris Placantonakis

Abstract:

Background: Glioblastoma (GBM) is the most common and deadly primary brain malignancy in adults. Tumor propagation, brain invasion, and resistance to therapy critically depend on GBM stem-like cells (GSCs); however, the mechanisms that regulate GSC self-renewal are incompletely understood. Given the aggressiveness and poor prognosis of GBM, it is imperative to find biomarkers that could also translate into novel drug targets. Along these lines, we have identified a cell surface antigen, CD97 (ADGRE5), an adhesion G protein-coupled receptor (GPCR), that is expressed on GBM cells but is absent from non-neoplastic brain tissue. CD97 has been shown to promote invasiveness, angiogenesis, and migration in several human cancers, but its frequency of expression and functional role in regulating GBM growth and survival, and its potential as a therapeutic target has not been investigated. Design: We assessed CD97 mRNA and protein expression in patient derived GBM samples and cell lines using publicly available RNA-sequencing datasets and flow cytometry, respectively. To assess CD97 function, we generated shRNA lentiviral constructs that target a sequence in the CD97 extracellular domain (ECD). A scrambled shRNA (scr) with no predicted targets in the genome was used as a control. We evaluated CD97 shRNA lentivirally transduced GBM cells for Ki67, Annexin V, and DAPI. We also tested CD97 KD cells for their ability to self-renew using clonogenic tumorsphere formation assays. Further, we utilized synthetic Abs (sAbs) generated against the ECD of CD97 to test for potential antitumor effects using patient-derived GBM cell lines. Results: CD97 mRNA expression was expressed at high levels in all GBM samples available in the TCGA cohort. We found high levels of surface CD97 protein expression in 6/6 patient-derived GBM cell cultures, but not human neural stem cells. Flow cytometry confirmed downregulation of CD97 in CD97 shRNA lentivirally transduced cells. CD97 KD induced a significant reduction in cell growth in 3 independent GBM cell lines representing mesenchymal and proneural subtypes, which was accompanied by reduced (~20%) Ki67 staining and increased (~30%) apoptosis. Incubation of GBM cells with sAbs (20 ug/ ml) against the ECD of CD97 for 3 days induced GSC differentiation, as determined by the expression of GFAP and Tubulin. Using three unique GBM patient derived cultures, we found that CD97 KD attenuated the ability of GBM cells to initiate sphere formation by over 300 fold, consistent with an impairment in GSC self-renewal. Conclusion: Loss of CD97 expression in patient-derived GBM cells markedly decreases proliferation, induces cell death, and reduces tumorsphere formation. sAbs against the ECD of CD97 reduce tumorsphere formation, recapitulating the phenotype of CD97 KD, suggesting that sAbs that inhibit CD97 function exhibit anti-tumor activity. Collectively, these findings indicate that CD97 is necessary for the proliferation and survival of human GBM cells and identify CD97 as a promising therapeutically targetable vulnerability in GBM.

Keywords: adhesion GPCR, CD97, GBM stem cell, glioblastoma

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12643 Design of a Controlled BHJ Solar Cell Using Modified Organic Vapor Spray Deposition Technique

Authors: F. Stephen Joe, V. Sathya Narayanan, V. R. Sanal Kumar

Abstract:

A comprehensive review of the literature on photovoltaic cells has been carried out for exploring the better options for cost efficient technologies for future solar cell applications. Literature review reveals that the Bulk Heterojunction (BHJ) Polymer Solar cells offer special opportunities as renewable energy resources. It is evident from the previous studies that the device fabricated with TiOx layer shows better power conversion efficiency than that of the device without TiOx layer. In this paper, authors designed a controlled BHJ solar cell using a modified organic vapor spray deposition technique facilitated with a vertical-moving gun named as 'Stephen Joe Technique' for getting a desirable surface pattern over the substrate to improving its efficiency over the years for industrial applications. We comprehended that the efficient processing and the interface engineering of these solar cells could increase the efficiency up to 5-10 %.

Keywords: BHJ polymer solar cell, photovoltaic cell, solar cell, Stephen Joe technique

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12642 Thermodynamic and Immunochemical Studies of Antibody Biofunctionalized Gold Nanoparticles Mediated Photothermal Ablation in Human Liver Cancer Cells

Authors: Lucian Mocan, Flaviu Tabaran, Teodora Mocan, Cristian Matea, Cornel Iancu

Abstract:

We present method of Gold Nanoparticle enhanced laser thermal ablation of HepG2 cells (Human hepatocellular liver carcinoma cell line), based on a simple gold nanoparticle carrier system, such as serum albumin (BSA), and demonstrate its selective therapeutic efficacy. Hyperspectral, contrast phase, and confocal microscopy combined immunochemical staining were used to demonstrate the selective internalization of HSA-GNPs via Gp60 receptors and the caveolin-mediated endocytosis inside HepG2 cells. We examined the ability of laser-activated carbon nanotubes to induce Hsp70 expression using confocal microscopy. Hep G2 cells heat-shocked (laser activated BSA-GNPs) to 42°C demonstrated an up-regulation of Hsp70 compared with control cells (BSA-GNPs treated cells without laser), which showed no detectable constitutive expression of Hsp70. We observed a time-dependent induction in Hsp70 expression in Hep G2 treated with BSA-GNPs and LASER irradiated. The post-irradiation apoptotic rate of HepG2 cells treated with HSA-GNPs ranged from 88.24% (for 50 mg/L) at 60 seconds, while at 30 minute the rate increased to 92.34% (50 mg/L). These unique results may represent a major step in liver cancer treatment using nanolocalized thermal ablation by laser heating.

Keywords: gold nanoparticles, liver cancer, albumin, laser irradiation

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12641 Functional Cell Surface Display Using Ice Nucleation Protein from Erwina ananas on Escherischia coli

Authors: Mei Yuin Joanne Wee, Rosli Md. Illias

Abstract:

Cell surface display is the expression of a protein with an anchoring motif on the surface of the cell. This approach offers advantages when used in bioconversion in terms of easier purification steps and more efficient enzymatic reaction. A surface display system using ice nucleation protein (InaA) from Erwina ananas as an anchoring motif has been constructed to display xylanase (xyl) on the surface of Escherischia coli. The InaA was truncated so that it is made up of the N- and C-terminal domain (INPANC-xyl) and it has successfully directed xylanase to the surface of the cell. A study was also done on xylanase fused to two other ice nucleation proteins, InaK (INPKNC-xyl) and InaZ (INPZNC-xyl) from Pseudomonas syringae KCTC 1832 and Pseudomonas syringae S203 respectively. Surface localization of the fusion protein was verified using SDS-PAGE and Western blot on the cell fractions and all anchoring motifs were successfully displayed on the outer membrane of E. coli. Upon comparison, whole-cell activity of INPANC-xyl was more than six and five times higher than INPKNC-xyl and INPZNC-xyl respectively. Furthermore, the expression of INPANC-xyl on the surface of E. coli did not inhibit the growth of the cell. This is the first report of surface display system using ice nucleation protein, InaA from E. ananas. From this study, this anchoring motif offers an attractive alternative to the current surface display systems.

Keywords: cell surface display, Escherischia coli, ice nucleation protein, xylanase

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12640 Catalytic Study of Natural Gas Based Solid Oxide Fuel Cell

Authors: Nasir Iqbal, Khurram Siraj, Rizwan Raza

Abstract:

Solid oxide fuel cell (SOFC) is the promising technology now days. SOFC can be operated with different types of fuels available. In this work catalytic anode is prepared with metal oxides i.e. Li, Ni, Zn and Sn and tested for catalytic activity with natural gas as a fuel. The operating temperature range is 170-750°C as observed with the help of TGA. Electrical conductivity and fuel cell performance has been observed for four different samples with varying composition of Sn and Zn. It is concluded that the sample having greater concentration of Zn shows better conductivity and power density results. All the results are promising and verified with different characterizations.

Keywords: catalytic activity, solid oxide fuel cell, energy material, natural gas

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12639 Effect of Plasma Radiation on Keratinocyte Cells Involved in the Wound Healing Process

Authors: B. Fazekas, I. Korolov, K. Kutasi

Abstract:

Plasma medicine, which involves the use of gas discharge plasmas for medical applications is a rapidly growing research field. The use of non-thermal atmospheric pressure plasmas in dermatology to assist tissue regeneration by improving the healing of infected and/or chronic wounds is a promising application. It is believed that plasma can activate cells, which are involved in the wound closure. Non-thermal atmospheric plasmas are rich in chemically active species (such as O and N-atoms, O2(a) molecules) and radiative species such as the NO, N2+ and N2 excited molecules, which dominantly radiate in the 200-500 nm spectral range. In order to understand the effect of plasma species, both of chemically active and radiative species on wound healing process, the interaction of physical plasma with the human skin cells is necessary. In order to clarify the effect of plasma radiation on the wound healing process we treated keratinocyte cells – that are one of the main cell types in human skin epidermis – covered with a layer of phosphate-buffered saline (PBS) with a low power atmospheric pressure plasma. For the generation of such plasma we have applied a plasma needle. Here, the plasma is ignited at the tip of the needle in flowing helium gas in contact with the ambient air. To study the effect of plasma radiation we used a plasma needle configuration, where the plasma species – chemically active radicals and charged species – could not reach the treated cells, but only the radiation. For the comparison purposes, we also irradiated the cells using a UV-B light source (FS20 lamp) with a 20 and 40 mJ cm-2 dose of 312 nm. After treatment the viability and the proliferation of the cells have been examined. The proliferation of cells has been studied with a real time monitoring system called Xcelligence. The results have indicated, that the 20 mJ cm-2 dose did not affect cell viability, whereas the 40 mJ cm-2 dose resulted a decrease in cell viability. The results have shown that the plasma radiation have no quantifiable effect on the cell proliferation as compared to the non-treated cells.

Keywords: UV radiation, non-equilibrium gas discharges (non-thermal plasmas), plasma emission, keratinocyte cells

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12638 Force Measurement for E-Cadherin-Mediated Intercellular Adhesion Probed by Protein Micropattern and Traction Force Microscopy

Authors: Chieh-Chung Tsou, Chun-Min Lo, Yeh-Shiu Chu

Abstract:

Cell’s mechanical forces provide important physical cues in regulation of proper cellular functions, such as cell differentiation, proliferation and migration. It is believed that adhesive forces generated by cell-cell interaction are able to transmit to the interior of cell through filamentous cortical cytoskeleton. Prominent among other membrane receptors, Cadherins are prototypical adhesive molecules able to generate remarkable forces to regulate intercellular adhesion. However, the mechanistic steps of mechano-transduction in Cadherin-mediated adhesion remain very controversial. We are interested in understanding how Cadherin protein complexes enable force generation and transmission at cell-cell contact in the initial stage of intercellular adhesion. For providing a better control of time, space, and substrate stiffness, in this study, a combination of protein micropattern, micropipette manipulation, and traction force microscopy is used. Pair micropattern with different forms confines cell spreading area and the gaps in pairs varied from 2 to 8 microns are applied for monitoring the forces that cell pairs generated, measured by traction force microscopy. Moreover, cell clones obtained from epithelial cells undergone genome editing are used to score the importance for known components of Cadherin complexes in force generation. We believe that our results from this combinatory mechanobiological method will provide deep insights on understanding the biophysical principle governing mechano- transduction of Cadherin-mediated intercellular adhesion.

Keywords: cadherin, intercellular adhesion, protein micropattern, traction force microscopy

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12637 Bioreactor for Cell-Based Impedance Measuring with Diamond Coated Gold Interdigitated Electrodes

Authors: Roman Matejka, Vaclav Prochazka, Tibor Izak, Jana Stepanovska, Martina Travnickova, Alexander Kromka

Abstract:

Cell-based impedance spectroscopy is suitable method for electrical monitoring of cell activity especially on substrates that cannot be easily inspected by optical microscope (without fluorescent markers) like decellularized tissues, nano-fibrous scaffold etc. Special sensor for this measurement was developed. This sensor consists of corning glass substrate with gold interdigitated electrodes covered with diamond layer. This diamond layer provides biocompatible non-conductive surface for cells. Also, a special PPFC flow cultivation chamber was developed. This chamber is able to fix sensor in place. The spring contacts are connecting sensor pads with external measuring device. Construction allows real-time live cell imaging. Combining with perfusion system allows medium circulation and generating shear stress stimulation. Experimental evaluation consist of several setups, including pure sensor without any coating and also collagen and fibrin coating was done. The Adipose derived stem cells (ASC) and Human umbilical vein endothelial cells (HUVEC) were seeded onto sensor in cultivation chamber. Then the chamber was installed into microscope system for live-cell imaging. The impedance measurement was utilized by vector impedance analyzer. The measured range was from 10 Hz to 40 kHz. These impedance measurements were correlated with live-cell microscopic imaging and immunofluorescent staining. Data analysis of measured signals showed response to cell adhesion of substrates, their proliferation and also change after shear stress stimulation which are important parameters during cultivation. Further experiments plan to use decellularized tissue as scaffold fixed on sensor. This kind of impedance sensor can provide feedback about cell culture conditions on opaque surfaces and scaffolds that can be used in tissue engineering in development artificial prostheses. This work was supported by the Ministry of Health, grants No. 15-29153A and 15-33018A.

Keywords: bio-impedance measuring, bioreactor, cell cultivation, diamond layer, gold interdigitated electrodes, tissue engineering

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12636 Usage of Cord Blood Stem Cells of Asphyxia Infants for Treatment

Authors: Ahmad Shah Farhat

Abstract:

Background: Prenatal asphyxia or birth asphyxia is the medical situation resulting from a newborn infant that lasts long enough during the birth process to cause physical harm, usually to the brain. Human umbilical cord blood (UCB) is a well-established source of hematopoietic stem/progenitor cells (HSPCs) for allogeneic stem cell transplantation. These can be used clinically to care for children with malignant diseases. Low O2 can cause in proliferation and differentiation of stem cells. Method: the cord blood of 11 infants with 3-5 Apgar scores or need to cardiac pulmonary Resuscitation as an asphyxia group and ten normal infants with more than 8 Apgar scores as the normal group was collected, and after isolating hematopoietic stem cells, the cells were cultured in enriched media for 14 days to compare the numbers of colonies by microscope. Results: There was a significant difference in the number of RBC precursor colonies (red colonies) in cultured media with 107 cord blood hematopoietic stem cells of infants who were exposed to hypoxemia in two wells of palate. There was not a significant difference in the number of white cell colonies in the two groups in the two wells of the plate. Conclusion: Hypoxia in the perinatal period can cause the increase of hematopoietic stem cells of cord blood, special red precursor stem cells in vitro, like an increase of red blood cells in the body when exposed to low oxygen conditions. Thus, it will be usable.

Keywords: asphyxia, neonre, stem cell, red cell

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12635 Continuum-Based Modelling Approaches for Cell Mechanics

Authors: Yogesh D. Bansod, Jiri Bursa

Abstract:

The quantitative study of cell mechanics is of paramount interest since it regulates the behavior of the living cells in response to the myriad of extracellular and intracellular mechanical stimuli. The novel experimental techniques together with robust computational approaches have given rise to new theories and models, which describe cell mechanics as a combination of biomechanical and biochemical processes. This review paper encapsulates the existing continuum-based computational approaches that have been developed for interpreting the mechanical responses of living cells under different loading and boundary conditions. The salient features and drawbacks of each model are discussed from both structural and biological points of view. This discussion can contribute to the development of even more precise and realistic computational models of cell mechanics based on continuum approaches or on their combination with microstructural approaches, which in turn may provide a better understanding of mechanotransduction in living cells.

Keywords: cell mechanics, computational models, continuum approach, mechanical models

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12634 Analysis of Performance of 3T1D Dynamic Random-Access Memory Cell

Authors: Nawang Chhunid, Gagnesh Kumar

Abstract:

On-chip memories consume a significant portion of the overall die space and power in modern microprocessors. On-chip caches depend on Static Random-Access Memory (SRAM) cells and scaling of technology occurring as per Moore’s law. Unfortunately, the scaling is affecting stability, performance, and leakage power which will become major problems for future SRAMs in aggressive nanoscale technologies due to increasing device mismatch and variations. 3T1D Dynamic Random-Access Memory (DRAM) cell is a non-destructive read DRAM cell with three transistors and a gated diode. In 3T1D DRAM cell gated diode (D1) acts as a storage device and also as an amplifier, which leads to fast read access. Due to its high tolerance to process variation, high density, and low cost of memory as compared to 6T SRAM cell, it is universally used by the advanced microprocessor for on chip data and program memory. In the present paper, it has been shown that 3T1D DRAM cell can perform better in terms of fast read access as compared to 6T, 4T, 3T SRAM cells, respectively.

Keywords: DRAM Cell, Read Access Time, Retention Time, Average Power dissipation

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12633 Diagnosis Of Static, Dynamic, And Mixed Eccentricity In Line Start Permanent Magnet Synchronous Motor By Using FEM

Authors: Mohamed Moustafa Mahmoud Sedky

Abstract:

In line start permanent magnet synchronous motor, eccentricity is a common fault that can make it necessary to remove the motor from the production line. However, because the motor may be inaccessible, diagnosing the fault is not easy. This paper presents an FEM that identifies different models, static eccentricity, dynamic eccentricity, and mixed eccentricity, at no load and full load. The method overcomes the difficulty of applying FEMs to transient behavior. It simulates motor speed, torque and flux density distribution along the air gap for SE, DE, and ME. This paper represents the various effects of different eccentricities types on the transient performance.

Keywords: line start permanent magnet, synchronous machine, static eccentricity, dynamic eccentricity, mixed eccentricity

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12632 In silico Repopulation Model of Various Tumour Cells during Treatment Breaks in Head and Neck Cancer Radiotherapy

Authors: Loredana G. Marcu, David Marcu, Sanda M. Filip

Abstract:

Advanced head and neck cancers are aggressive tumours, which require aggressive treatment. Treatment efficiency is often hindered by cancer cell repopulation during radiotherapy, which is due to various mechanisms triggered by the loss of tumour cells and involves both stem and differentiated cells. The aim of the current paper is to present in silico simulations of radiotherapy schedules on a virtual head and neck tumour grown with biologically realistic kinetic parameters. Using the linear quadratic formalism of cell survival after radiotherapy, altered fractionation schedules employing various treatment breaks for normal tissue recovery are simulated and repopulation mechanism implemented in order to evaluate the impact of various cancer cell contribution on tumour behaviour during irradiation. The model has shown that the timing of treatment breaks is an important factor influencing tumour control in rapidly proliferating tissues such as squamous cell carcinomas of the head and neck. Furthermore, not only stem cells but also differentiated cells, via the mechanism of abortive division, can contribute to malignant cell repopulation during treatment.

Keywords: radiation, tumour repopulation, squamous cell carcinoma, stem cell

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12631 Comparison between the Efficiency of Heterojunction Thin Film InGaP\GaAs\Ge and InGaP\GaAs Solar Cell

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

Abstract:

This paper presents the design parameters for a thin film 3J InGaP/GaAs/Ge solar cell with a simulated maximum efficiency of 32.11% using Tcad Silvaco. Design parameters include the doping concentration, molar fraction, layers’ thickness and tunnel junction characteristics. An initial dual junction InGaP/GaAs model of a previous published heterojunction cell was simulated in Tcad Silvaco to accurately predict solar cell performance. To improve the solar cell’s performance, we have fixed meshing, material properties, models and numerical methods. However, thickness and layer doping concentration were taken as variables. We, first simulate the InGaP\GaAs dual junction cell by changing the doping concentrations and thicknesses which showed an increase in efficiency. Next, a triple junction InGaP/GaAs/Ge cell was modeled by adding a Ge layer to the previous dual junction InGaP/GaAs model with an InGaP /GaAs tunnel junction.

Keywords: heterojunction, modeling, simulation, thin film, Tcad Silvaco

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12630 The Operating Behaviour of Unbalanced Unpaced Merging Assembly Lines

Authors: S. Shaaban, T. McNamara, S. Hudson

Abstract:

This paper reports on the performance of deliberately unbalanced, reliable, non-automated and assembly lines that merge, whose workstations differ in terms of their mean operation times. Simulations are carried out on 5- and 8-station lines with 1, 2 and 4 buffer capacity units, % degrees of line imbalance of 2, 5 and 12, and 24 different patterns of means imbalance. Data on two performance measures, namely throughput and average buffer level were gathered, statistically analysed and compared to a merging balanced line counterpart. It was found that the best configurations are a balanced line arrangement and a monotone decreasing order for each of the parallel merging lines, with the first generally resulting in a lower throughput and the second leading to a lower average buffer level than those of a balanced line.

Keywords: average buffer level, merging lines, simulation, throughput, unbalanced

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12629 Anti-proliferative Activity and HER2 Receptor Expression Analysis of MCF-7 (Breast Cancer Cell) Cells by Plant Extract Coleus Barbatus (Andrew)

Authors: Anupalli Roja Rani, Pavithra Dasari

Abstract:

Background: Among several, breast cancer has emerged as the most common female cancer in developing countries. It is the most common cause of cancer-related deaths worldwide among women. It is a molecularly and clinically heterogeneous disease. Moreover, it is a hormone–dependent tumor in which estrogens can regulate the growth of breast cells by binding with estrogen receptors (ERs). Moreover, the use of natural products in cancer therapeutics is due to their properties of biocompatibility and less toxicity. Plants are the vast reservoirs for various bioactive compounds. Coleus barbatus (Lamiaceae) contains anticancer properties against several cancer cell lines. Method: In the present study, an attempt is being made to enrich the knowledge of the anticancer activity of pure compounds extracted from Coleus barbatus (Andrew). On human breast cancer cell lines MCF-7. Here in, we are assessing the antiproliferative activity of Coleus barbatus (Andrew) plant extracts against MCF 7 and also evaluating their toxicity in normal human mammary cell lines such as Human Mammary Epithelial Cells (HMEC). The active fraction of plant extract was further purified with the help of Flash chromatography, Medium Pressure Liquid Chromatography (MPLC) and preparative High-Performance Liquid Chromatography (HPLC). The structure of pure compounds will be elucidated by using modern spectroscopic methods like Nuclear magnetic resonance (NMR), Electrospray Ionisation Mass Spectrometry (ESI-MS) methods. Later, the growth inhibition morphological assessment of cancer cells and cell cycle analysis of purified compounds were assessed using FACS. The growth and progression of signaling molecules HER2, GRP78 was studied by secretion assay using ELISA and expression analysis by flow cytometry. Result: Cytotoxic effect against MCF-7 with IC50 values were derived from dose response curves, using six concentrations of twofold serially diluted samples, by SOFTMax Pro software (Molecular device) and respectively Ellipticine and 0.5% DMSO were used as a positive and negative control. Conclusion: The present study shows the significance of various bioactive compounds extracted from Coleus barbatus (Andrew) root material. It acts as an anti-proliferative and shows cytotoxic effects on human breast cancer cell lines MCF7. The plant extracts play an important role pharmacologically. The whole plant has been used in traditional medicine for decades and the studies done have authenticated the practice. Earlier, as described, the plant has been used in the ayurveda and homeopathy medicine. However, more clinical and pathological studies must be conducted to investigate the unexploited potential of the plant. These studies will be very useful for drug designing in the future.

Keywords: coleus barbatus, HPLC, MPLC, NMR, MCF7, flash chromatograph, ESI-MS, FACS, ELISA.

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12628 The Stem Cell Transcription Co-factor Znf521 Sustains Mll-af9 Fusion Protein In Acute Myeloid Leukemias By Altering The Gene Expression Landscape

Authors: Emanuela Chiarella, Annamaria Aloisio, Nisticò Clelia, Maria Mesuraca

Abstract:

ZNF521 is a stem cell-associated transcription co-factor, that plays a crucial role in the homeostatic regulation of the stem cell compartment in the hematopoietic, osteo-adipogenic, and neural system. In normal hematopoiesis, primary human CD34+ hematopoietic stem cells display typically a high expression of ZNF521, while its mRNA levels rapidly decrease when these progenitors progress towards erythroid, granulocytic, or B-lymphoid differentiation. However, most acute myeloid leukemias (AMLs) and leukemia-initiating cells keep high ZNF521 expression. In particular, AMLs are often characterized by chromosomal translocations involving the Mixed Lineage Leukemia (MLL) gene, which MLL gene includes a variety of fusion oncogenes arisen from genes normally required during hematopoietic development; once they are fused, they promote epigenetic and transcription factor dysregulation. The chromosomal translocation t(9;11)(p21-22;q23), fusing the MLL gene with AF9 gene, results in a monocytic immune phenotype with an aggressive course, frequent relapses, and a short survival time. To better understand the dysfunctional transcriptional networks related to genetic aberrations, AML gene expression profile datasets were queried for ZNF521 expression and its correlations with specific gene rearrangements and mutations. The results showed that ZNF521 mRNA levels are associated with specific genetic aberrations: the highest expression levels were observed in AMLs involving t(11q23) MLL rearrangements in two distinct datasets (MILE and den Boer); elevated ZNF521 mRNA expression levels were also revealed in AMLs with t(7;12) or with internal rearrangements of chromosome 16. On the contrary, relatively low ZNF521 expression levels seemed to be associated with the t(8;21) translocation, that in turn is correlated with the AML1-ETO fusion gene or the t(15;17) translocation and in AMLs with FLT3-ITD, NPM1, or CEBPα double mutations. Invitro, we found that the enforced co-expression of ZNF521 in cord blood-derived CD34+ cells induced a significant proliferative advantage, improving MLL-AF9 effects on the induction of proliferation and the expansion of leukemic progenitor cells. Transcriptome profiling of CD34+ cells transduced with either MLL-AF9, ZNF521, or a combination of the two transgenes highlighted specific sets of up- or down-regulated genes that are involved in the leukemic phenotype, including those encoding transcription factors, epigenetic modulators, and cell cycle regulators as well as those engaged in the transport or uptake of nutrients. These data enhance the functional cooperation between ZNF521 and MA9, resulting in the development, maintenance, and clonal expansion of leukemic cells. Finally, silencing of ZNF521 in MLL-AF9-transformed primary CD34+ cells inhibited their proliferation and led to their extinction, as well as ZNF521 silencing in the MLL-AF9+ THP-1 cell line resulted in an impairment of their growth and clonogenicity. Taken together, our data highlight ZNF521 role in the control of self-renewal and in the immature compartment of malignant hematopoiesis, which, by altering the gene expression landscape, contributes to the development and/or maintenance of AML acting in concert with the MLL-AF9 fusion oncogene.

Keywords: AML, human zinc finger protein 521 (hZNF521), mixed lineage leukemia gene (MLL) AF9 (MLLT3 or LTG9), cord blood-derived hematopoietic stem cells (CB-CD34+)

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12627 Safety Study of Intravenously Administered Human Cord Blood Stem Cells in the Treatment of Symptoms Related to Chronic Inflammation

Authors: Brian M. Mehling, Louis Quartararo, Marine Manvelyan, Paul Wang, Dong-Cheng Wu

Abstract:

Numerous investigations suggest that Mesenchymal Stem Cells (MSCs) in general represent a valuable tool for therapy of symptoms related to chronic inflammatory diseases. Blue Horizon Stem Cell Therapy Program is a leading provider of adult and children’s stem cell therapies. Uniquely we have safely and efficiently treated more than 600 patients with documenting each procedure. The purpose of our study is primarily to monitor the immune response in order to validate the safety of intravenous infusion of human umbilical cord blood derived MSCs (UC-MSCs), and secondly, to evaluate effects on biomarkers associated with chronic inflammation. Nine patients were treated for conditions associated with chronic inflammation and for the purpose of anti-aging. They have been given one intravenous infusion of UC-MSCs. Our study of blood test markers of 9 patients with chronic inflammation before and within three months after MSCs treatment demonstrates that there is no significant changes and MSCs treatment was safe for the patients. Analysis of different indicators of chronic inflammation and aging included in initial, 24-hours, two weeks and three months protocols showed that stem cell treatment was safe for the patients; there were no adverse reactions. Moreover data from follow up protocols demonstrates significant improvement in energy level, hair, nails growth and skin conditions. Intravenously administered UC-MSCs were safe and effective in the improvement of symptoms related to chronic inflammation. Further close monitoring and inclusion of more patients are necessary to fully characterize the advantages of UC-MSCs application in treatment of symptoms related to chronic inflammation.

Keywords: chronic inflammatory diseases, intravenous infusion, stem cell therapy, umbilical cord blood derived mesenchymal stem cells (UC-MSCs)

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12626 Optimizing Cell Culture Performance in an Ambr15 Microbioreactor Using Dynamic Flux Balance and Computational Fluid Dynamic Modelling

Authors: William Kelly, Sorelle Veigne, Xianhua Li, Zuyi Huang, Shyamsundar Subramanian, Eugene Schaefer

Abstract:

The ambr15™ bioreactor is a single-use microbioreactor for cell line development and process optimization. The ambr system offers fully automatic liquid handling with the possibility of fed-batch operation and automatic control of pH and oxygen delivery. With operating conditions for large scale biopharmaceutical production properly scaled down, micro bioreactors such as the ambr15™ can potentially be used to predict the effect of process changes such as modified media or different cell lines. In this study, gassing rates and dilution rates were varied for a semi-continuous cell culture system in the ambr15™ bioreactor. The corresponding changes to metabolite production and consumption, as well as cell growth rate and therapeutic protein production were measured. Conditions were identified in the ambr15™ bioreactor that produced metabolic shifts and specific metabolic and protein production rates also seen in the corresponding larger (5 liter) scale perfusion process. A Dynamic Flux Balance model was employed to understand and predict the metabolic changes observed. The DFB model-predicted trends observed experimentally, including lower specific glucose consumption when CO₂ was maintained at higher levels (i.e. 100 mm Hg) in the broth. A Computational Fluid Dynamic (CFD) model of the ambr15™ was also developed, to understand transfer of O₂ and CO₂ to the liquid. This CFD model predicted gas-liquid flow in the bioreactor using the ANSYS software. The two-phase flow equations were solved via an Eulerian method, with population balance equations tracking the size of the gas bubbles resulting from breakage and coalescence. Reasonable results were obtained in that the Carbon Dioxide mass transfer coefficient (kLa) and the air hold up increased with higher gas flow rate. Volume-averaged kLa values at 500 RPM increased as the gas flow rate was doubled and matched experimentally determined values. These results form a solid basis for optimizing the ambr15™, using both CFD and FBA modelling approaches together, for use in microscale simulations of larger scale cell culture processes.

Keywords: cell culture, computational fluid dynamics, dynamic flux balance analysis, microbioreactor

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12625 An Investigation of Tetraspanin Proteins’ Role in UPEC Infection

Authors: Fawzyah Albaldi

Abstract:

Urinary tract infections (UTIs) are the most prevalent of infectious diseases and > 80% are caused by uropathogenic E. coli (UPEC). Infection occurs following adhesion to urothelial plaques on bladder epithelial cells, whose major protein constituent are the uroplakins (UPs). Two of the four uroplakins (UPIa and UPIb) are members of the tetraspanin superfamily. The UPEC adhesin FimH is known to interact directly with UPIa. Tetraspanins are a diverse family of transmembrane proteins that generally act as “molecular organizers” by binding different proteins and lipids to form tetraspanin enriched microdomains (TEMs). Previous work by our group has shown that TEMs are involved in the adhesion of many pathogenic bacteria to human cells. Adhesion can be blocked by tetraspanin-derived synthetic peptides, suggesting that tetraspanins may be valuable drug targets. In this study, we investigate the role of tetraspanins in UPEC adherence to bladder epithelial cells. Human bladder cancer cell lines (T24, 5637, RT4), commonly used as in-vitro models to investigate UPEC infection, along with primary human bladder cells, were used in this project. The aim was to establish a model for UPEC adhesion/infection with the objective of evaluating the impact of tetraspanin-derived reagents on this process. Such reagents could reduce the progression of UTI, particularly in patients with indwelling catheters. Tetraspanin expression on the bladder cells was investigated by q-PCR and flow cytometry, with CD9 and CD81 generally highly expressed. Interestingly, despite these cell lines being used by other groups to investigate FimH antagonists, uroplakin proteins (UPIa, UPIb and UPIII) were poorly expressed at the cell surface, although some were present intracellularly. Attempts were made to differentiate the cell lines, to induce cell surface expression of these UPs, but these were largely unsuccessful. Pre-treatment of bladder epithelial cells with anti-CD9 monoclonal antibody significantly decreased UPEC infection, whilst anti-CD81 had no effects. A short (15aa) synthetic peptide corresponding to the large extracellular region (EC2) of CD9 also significantly reduced UPEC adherence. Furthermore, we demonstrated specific binding of that fluorescently tagged peptide to the cells. CD9 is known to associate with a number of heparan sulphate proteoglycans (HSPGs) that have also been implicated in bacterial adhesion. Here, we demonstrated that unfractionated heparin (UFH)and heparin analogs significantly inhibited UPEC adhesion to RT4 cells, as did pre-treatment of the cells with heparinases. Pre-treatment with chondroitin sulphate (CS) and chondroitinase also significantly decreased UPEC adherence to RT4 cells. This study may shed light on a common pathogenicity mechanism involving the organisation of HSPGs by tetraspanins. In summary, although we determined that the bladder cell lines were not suitable to investigate the role of uroplakins in UPEC adhesion, we demonstrated roles for CD9 and cell surface proteoglycans in this interaction. Agents that target these may be useful in treating/preventing UTIs.

Keywords: UTIs, tspan, uroplakins, CD9

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12624 Numerical Simulation of Multijunction GaAs/CIGS Solar Cell by AMPS-1D

Authors: Hassane Ben Slimane, Benmoussa Dennai, Abderrahman Hemmani, Abderrachid Helmaoui

Abstract:

During the past few years a great variety of multi-junction solar cells has been developed with the aim of a further increase in efficiency beyond the limits of single junction devices. This paper analyzes the GaAs/CIGS based tandem solar cell performance by AMPS-1D numerical modeling. Various factors which affect the solar cell’s performance are investigated, carefully referring to practical cells, to obtain the optimum parameters for the GaAs and CIGS top and bottom solar cells. Among the factors studied are thickness and band gap energy of dual junction cells.

Keywords: multijunction solar cell, GaAs, CIGS, AMPS-1D

Procedia PDF Downloads 483
12623 Human Capital Development: A Pivotal for Sustainable Development in Developing Countries

Authors: Yusuf Ismaila

Abstract:

The developing countries are characterized by inefficient production systems and unequal distribution of wealth. Developing countries are largely populated, yet under developed. This can be attributed partly to the unplanned efforts towards the development of human capital through education and training. In the developed nations a huge attention is accorded to indices such as life expectancy, literacy, infant mortality, education, and the efficient delivery of social services. This is the reason why many developing countries have been scored low by the United Nations in terms of its human development indicators. The population growth continued to expand far beyond the rate of economic growth, a situation that gave rise to increasing poverty. This paper examines the effect of selected human development indicators on the economic development. Thus human capital development is one of the fundamental solutions to enter the international arena. Both quantitative and qualitative analyses were used to demonstrate the effect of selected human capital indices and related literatures were also reviewed for exposition of the human capital concept. It was found that there are no conscious efforts in human capital planning. This has therefore resulted to continuing dwindling of production system and poverty. Recommendations made to redress the situation include that human capital development should be planned and adequately funded in line with the needs of the economy and by applying international standards. Specifically, developing countries must invest necessary resources in developing human capital which tend to have a great impact on sustainable development. Information about the labour market should improve while government policy should favour labour mobility. HCD strategy must focus on improving the skills of the workforce, reducing the cost of doing business and making available the resources business needs to compete and thrive in a fast globalizing economy. There should be regular interaction of planners, employers and builders of human capital to facilitate the process of meaningful national development.

Keywords: economic development, human capital, economic growth, developing countries

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12622 Capture of Co₂ From Natural Gas Using Modified Imidazolium Ionic Liquids

Authors: Alaa A. Ghanem, S. E. M. Desouky

Abstract:

Natural gas (NG) is considered one of the most essential global energy sources. NG fields are often far away from the market, and a long-distance transporting pipeline usually is required. Production of NG with high content of CO₂ leads to severe problems such as equipment corrosion along with the production line until refinery.in addition to a high level of toxicity and decreasing in calorific value of the NG. So it is recommended to remove or decrease the CO₂ percent to meet transport specifications. This can be reached using different removal techniques such as physical and chemical absorption, pressure swing adsorption, membrane separation, or low-temperature separation. Many solvents and chemicals are being used to capture carbon dioxide on a large scale; among them, Ionic liquids have great potential due to their tunable properties; low vapour pressure, low melting point, and sensible thermal stability. In this research, three modifiedimidazolium ionic liquids will be synthesized and characterized using different tools of analysis such as FT-IR, 1H NMR. Thermal stability and surface activity will be studied. The synthesized compounds will be evaluated as selective solvents for CO₂ removal from natural gas using PVT cell.

Keywords: natural gas, CO₂ capture, imidazolium ionic liquid, PVT cell

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12621 A Study of the Alumina Distribution in the Lab-Scale Cell during Aluminum Electrolysis

Authors: Olga Tkacheva, Pavel Arkhipov, Alexey Rudenko, Yurii Zaikov

Abstract:

The aluminum electrolysis process in the conventional cryolite-alumina electrolyte with cryolite ratio of 2.7 was carried out at an initial temperature of 970 °C and the anode current density of 0.5 A/cm2 in a 15A lab-scale cell in order to study the formation of the side ledge during electrolysis and the alumina distribution between electrolyte and side ledge. The alumina contained 35.97% α-phase and 64.03% γ-phase with the particles size in the range of 10-120 μm. The cryolite ratio and the alumina concentration were determined in molten electrolyte during electrolysis and in frozen bath after electrolysis. The side ledge in the electrolysis cell was formed only by the 13th hour of electrolysis. With a slight temperature decrease a significant increase in the side ledge thickness was observed. The basic components of the side ledge obtained by the XRD phase analysis were Na3AlF6, Na5Al3F14, Al2O3, and NaF.5CaF2.AlF3. As in the industrial cell, the increased alumina concentration in the side ledge formed on the cell walls and at the ledge-electrolyte-aluminum three-phase boundary during aluminum electrolysis in the lab cell was found (FTP No 05.604.21.0239, IN RFMEFI60419X0239).

Keywords: alumina distribution, aluminum electrolyzer, cryolie-alumina electrolyte, side ledge

Procedia PDF Downloads 250
12620 Computational Approaches to Study Lineage Plasticity in Human Pancreatic Ductal Adenocarcinoma

Authors: Almudena Espin Perez, Tyler Risom, Carl Pelz, Isabel English, Robert M. Angelo, Rosalie Sears, Andrew J. Gentles

Abstract:

Pancreatic ductal adenocarcinoma (PDAC) is one of the most deadly malignancies. The role of the tumor microenvironment (TME) is gaining significant attention in cancer research. Despite ongoing efforts, the nature of the interactions between tumors, immune cells, and stromal cells remains poorly understood. The cell-intrinsic properties that govern cell lineage plasticity in PDAC and extrinsic influences of immune populations require technically challenging approaches due to the inherently heterogeneous nature of PDAC. Understanding the cell lineage plasticity of PDAC will improve the development of novel strategies that could be translated to the clinic. Members of the team have demonstrated that the acquisition of ductal to neuroendocrine lineage plasticity in PDAC confers therapeutic resistance and is a biomarker of poor outcomes in patients. Our approach combines computational methods for deconvolving bulk transcriptomic cancer data using CIBERSORTx and high-throughput single-cell imaging using Multiplexed Ion Beam Imaging (MIBI) to study lineage plasticity in PDAC and its relationship to the infiltrating immune system. The CIBERSORTx algorithm uses signature matrices from immune cells and stroma from sorted and single-cell data in order to 1) infer the fractions of different immune cell types and stromal cells in bulked gene expression data and 2) impute a representative transcriptome profile for each cell type. We studied a unique set of 300 genomically well-characterized primary PDAC samples with rich clinical annotation. We deconvolved the PDAC transcriptome profiles using CIBERSORTx, leveraging publicly available single-cell RNA-seq data from normal pancreatic tissue and PDAC to estimate cell type proportions in PDAC, and digitally reconstruct cell-specific transcriptional profiles from our study dataset. We built signature matrices and optimized by simulations and comparison to ground truth data. We identified cell-type-specific transcriptional programs that contribute to cancer cell lineage plasticity, especially in the ductal compartment. We also studied cell differentiation hierarchies using CytoTRACE and predict cell lineage trajectories for acinar and ductal cells that we believe are pinpointing relevant information on PDAC progression. Collaborators (Angelo lab, Stanford University) has led the development of the Multiplexed Ion Beam Imaging (MIBI) platform for spatial proteomics. We will use in the very near future MIBI from tissue microarray of 40 PDAC samples to understand the spatial relationship between cancer cell lineage plasticity and stromal cells focused on infiltrating immune cells, using the relevant markers of PDAC plasticity identified from the RNA-seq analysis.

Keywords: deconvolution, imaging, microenvironment, PDAC

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12619 The Impact of Artificial Intelligence on Human Rights Development and Obligations

Authors: Bola George Asaad Bekledas

Abstract:

Relationship between development and human rights has long been the subject of academic debate. To understand the dynamics between these two concepts, various principles are adopted, from the right to development to development-based human rights. Despite the initiatives taken, the relationship between development and human rights remains unclear. However, the compatibility between these two concepts and the idea that these efforts should be made to respect human rights guarantees have gained momentum in recent years. It is then evaluated whether the right to sustainable development is acceptable or not. The article concludes that the principles of sustainable development are directly or indirectly recognized in various human rights instruments, which is a good answer to the question posed above. This study, therefore, cites regional and international human rights agreements such as, as well as the jurisprudence and interpretative guidelines of human rights institutions, to prove this hypothesis.

Keywords: sustainable development, human rights, the right to development, the human rights-based approach to development, environmental rights, economic development, social sustainability human rights protection, human rights violations, workers’ rights, justice, security.

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12618 Isolation and Elimination of Latent and Productive Herpes Simplex Virus from the Sacral and Trigeminal Ganglions

Authors: Bernard L. Middleton, Susan P. Cosgrove

Abstract:

There is an immediate need for alternative anti-herpetic treatment options effective for both primary infections and reoccurring reactivations of herpes simplex virus types 1 (HSV-1) and 2 (HSV-2). Alternatives currently approved for the purposes of clinical administration includes antivirals and a reduced set of nucleoside analogues. The present article tests a treatment based on a systemic understanding of how the herpes virus affects cell inhibition and breakdown and targets different phases of the viral cycle, including the entry stage, reproductive cross mutation, and cell-to-cell infection. The treatment consisted of five immunotherapeutic core compounds (5CC), which were hypothesized to be capable of neutralizing human monoclonal antibodies. The tested 5CC were noted as being functional in the application of eliminating the DNA synthesis of herpes viral interferon (IFN) - induced cellular antiviral response. They were here found to neutralize antiviral reproduction by blocking cell-to-cell infection. The activity of the 5CC was tested on RC-37 in vitro using an assay plaque reduction and in vivo against HSV-1 and HSV-2. The 50% inhibitory concentration (IC50) of 5CC was 0.0009% for HSV-1 plaque formation and 0.0008% for HSV-2 plaque formation. Further tests were performed to evaluate the susceptibility of HSV-1 and HSV-2 to anti-herpetic drugs in Vero cells after virus entry. There were high-level markers of the 5CC virucidal activity in the viral suspension of HSV-1 and HSV-2. These concentrations of the 5CC are nontoxic and reduced plaque formation by 98.2% for HSV-1 and 93.0% for HSV-2. Virus HSV-1 and HSV-2 titers were reduced significantly by 5CC to the point of being negative, ranging 0.01–0.09 in 72%. The results concluded the 5CC as being an effective treatment option for the herpes simplex virus.

Keywords: synergy pharmaceuticals, herpes treatment, herpes cure, synergy pharmaceuticals treatment

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12617 Treatment of NMSC with Traditional Medicine Method

Authors: Aferdita Stroka Koka, Laver Stroka, Juna Musa, Samanda Celaj

Abstract:

Non-melanoma skin cancers (NMSCs) are the most common human malignancies. About 5.4 million basal and squamous cell skin cancers are diagnosed each year in the US and new cases continue to grow. About eight out of ten of these are basal cell cancers. Squamous cell cancers occur less often. NMSC usually are treatable, but treatment is expensive and can leave scars. In 2019, 167 patients of both sexes suffering from NMSC were treated by traditional medicine. Patients who have been diagnosed with Basal Cell Carcinoma were 122 cases, Squamous Cell Carcinoma 32 cases and both of them 13 cases. Of these,122 cases were ulcerated lesions and 45 unulcerated lesions. All patients were treated with the herbal solution called NILS, which contains extracts of some Albanian plants such as Allium sativum, Jugulans regia and Laurus nobilis. The treatment is done locally, on the surface of the tumor, applying the solution until the tumor mass is destroyed and, after that, giving the necessary time to the wound to make the regeneration that coincides with the complete healing of the wound. We have prepared a collection of photos for each case. Since the first sessions, a shrinkage and reduction of the tumor mass were evident, up to the total disappearance of the lesion at the end of treatment. The normal period of treatment lasted 1 to 2 weeks, depending on the size of the tumor, then take care of it until the closure of the wound, taking the whole process from 1 to 3 months. In 7 patients, the lesion failed to be dominated by treatment and they underwent standard treatment with radiotherapy or surgery, while in 10 patients, the lesion recurred and was treated again. The aim of this survey was to put in evidence the good results obtained by treatment of NMSC with Albanian traditional medicine methods.

Keywords: local treatment, nils, NMSC, traditional medicine

Procedia PDF Downloads 189
12616 Independent Control over Surface Charge and Wettability Using Polyelectrolyte Architecture

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

Abstract:

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

Procedia PDF Downloads 243