Search results for: cell concentration
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 8152

Search results for: cell concentration

7642 Long-Term Indoor Air Monitoring for Students with Emphasis on Particulate Matter (PM2.5) Exposure

Authors: Seyedtaghi Mirmohammadi, Jamshid Yazdani, Syavash Etemadi Nejad

Abstract:

One of the main indoor air parameters in classrooms is dust pollution and it depends on the particle size and exposure duration. However, there is a lake of data about the exposure level to PM2.5 concentrations in rural area classrooms. The objective of the current study was exposure assessment for PM2.5 for students in the classrooms. One year monitoring was carried out for fifteen schools by time-series sampling to evaluate the indoor air PM2.5 in the rural district of Sari city, Iran. A hygrometer and thermometer were used to measure some psychrometric parameters (temperature, relative humidity, and wind speed) and Real-Time Dust Monitor, (MicroDust Pro, Casella, UK) was used to monitor particulate matters (PM2.5) concentration. The results show the mean indoor PM2.5 concentration in the studied classrooms was 135µg/m3. The regression model indicated that a positive correlation between indoor PM2.5 concentration and relative humidity, also with distance from city center and classroom size. Meanwhile, the regression model revealed that the indoor PM2.5 concentration, the relative humidity, and dry bulb temperature was significant at 0.05, 0.035, and 0.05 levels, respectively. A statistical predictive model was obtained from multiple regressions modeling for indoor PM2.5 concentration and indoor psychrometric parameters conditions.

Keywords: classrooms, concentration, humidity, particulate matters, regression

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7641 New Targets Promoting Oncolytic Virotherapy

Authors: Felicia Segeth, Florian G. Klein, Lea Berger, Andreas Kolk, Per S. Holm

Abstract:

The entry of oncolytic viruses (OVs) into clinical application opens groundbreaking changes in current and future treatment regimens. However, despite their potent anti-cancer activity in vitro, clinical studies revealed limitations of OVs as monotherapy. The same applies to CDK 4/6 inhibitors (CDK4/6i) targeting cell cycle as well as bromodomain and extra-terminal domain inhibitors (BETi) targeting gene expression. In this study, the anti-tumoral effect of XVir-N-31, an YB-1 dependent oncolytic adenovirus, was evaluated in combination with Ribociclib, a CDK4/6i, and JQ1, a BETi. The head and neck squamous cell carcinoma (HNSCC) cell lines Fadu, SAS, and Cal-33 were used. DNA replication and gene expression of XVir-N-31 was measured by RT-qPCR, protein expression by western blotting, and cell lysis by SRB assays. Treatment with CDK4/6i and BETi increased viral gene expression, viral DNA replication, and viral particle formation. The data show that the combination of oncolytic adenovirus XVir-N-31 with CDK4/6i & BETi acts highly synergistic in cancer cell lysis. Furthermore, additional molecular analyses on this subject demonstrate that the positive transcription elongation factor P-TEFb plays a decisive role in this regard, indicating an influence of the combinational therapy on gene transcription control. The combination of CDK4/6i & BETi and XVir-N-31 is an attractive strategy to achieve substantial cancer cell killing and is highly suitable for clinical testing.

Keywords: adenovirus, BET, CDK4/6, HNSCC, P-TEFb, YB-1

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7640 Effect of SCN5A Gene Mutation in Endocardial Cell

Authors: Helan Satish, M. Ramasubba Reddy

Abstract:

The simulation of an endocardial cell for gene mutation in the cardiac sodium ion channel NaV1.5, encoded by SCN5A gene, is discussed. The characterization of Brugada Syndrome by loss of function effect on SCN5A mutation due to L812Q mutant present in the DII-S4 transmembrane region of the NaV1.5 channel protein and its effect in an endocardial cell is studied. Ten Tusscher model of human ventricular action potential is modified to incorporate the changes contributed by L812Q mutant in the endocardial cells. Results show that BrS-associated SCN5A mutation causes reduction in the inward sodium current by modifications in the channel gating dynamics such as delayed activation, enhanced inactivation, and slowed recovery from inactivation in the endocardial cell. A decrease in the inward sodium current was also observed, which affects depolarization phase (Phase 0) that leads to reduction in the spike amplitude of the cardiac action potential.

Keywords: SCN5A gene mutation, sodium channel, Brugada syndrome, cardiac arrhythmia, action potential

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7639 Solar Cell Using Chemical Bath Deposited PbS:Bi3+ Films as Electron Collecting Layer

Authors: Melissa Chavez Portillo, Mauricio Pacio Castillo, Hector Juarez Santiesteban, Oscar Portillo Moreno

Abstract:

Chemical bath deposited PbS:Bi3+ as an electron collection layer is introduced between the silicon wafer and the Ag electrode the performance of the PbS heterojunction thin film solar thin film solar cells with 1 cm2 active area. We employed Bi-doping to transform it into an n-type semiconductor. The experimental results reveal that the cell response parameters depend critically on the deposition procedures in terms of bath temperature, deposition time. The device achieves an open-circuit voltage of 0.4 V. The simple and low-cost deposition method of PbS:Bi3+ films is promising for the fabrication.

Keywords: Bi doping, PbS, thin films, solar cell

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7638 Determination of Heavy Metals (Cd, Pb, Hg, Cu, Fe, Mn, Al, As, Ni and Zn) in 6 Important Commercial Fish Species in North of Hormoz Strait

Authors: Majid Afkhami, Maryam Ehsanpour, Zahra Khoshnood

Abstract:

The concentrations of 10 heavy metals (Cd, Pb, Hg, Cu, Fe, Mn, Al, As, Ni, Zn) were measured in muscle, gill and liver of 6 species from Hormoz Strait in north coast of Persian Gulf in 12 months (April 2009 – March 2010). All samples were analyzed three times for Cd, Pb, Cu, Fe, Mn, Al, As, Ni, Zn by inductively coupled plasma-atomic emission spectrometry (ICP-AES) and for Hg by LECO AMA254 Advanced Mercury Analyzer. Results of this study showed that iron had the highest concentration (total mean concentration) in all species, followed by Zn, Cu, Ni, Al, Pb, Mn, Cd, Hg and lowest concentration in three tissues was As. In addition, the accumulation of metals was species-dependent, and was higher in Scomberomorous commerson and Scomberomorous guttatus (p<0.05) and the lowest concentration was record in Pampus argenteus (p<0.05).

Keywords: Persian Gulf, heavy metals, Hormoz strait, Scomberomorous guttatus, Scomberomorous commerson, Pampus argenteus

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7637 Incorporating Spatial Transcriptome Data into Ligand-Receptor Analyses to Discover Regional Activation in Cells

Authors: Eric Bang

Abstract:

Interactions between receptors and ligands are crucial for many essential biological processes, including neurotransmission and metabolism. Ligand-receptor analyses that examine cell behavior and interactions often utilize cell type-specific RNA expressions from single-cell RNA sequencing (scRNA-seq) data. Using CellPhoneDB, a public repository consisting of ligands, receptors, and ligand-receptor interactions, the cell-cell interactions were explored in a specific scRNA-seq dataset from kidney tissue and portrayed the results with dot plots and heat maps. Depending on the type of cell, each ligand-receptor pair was aligned with the interacting cell type and calculated the positori probabilities of these associations, with corresponding P values reflecting average expression values between the triads and their significance. Using single-cell data (sample kidney cell references), genes in the dataset were cross-referenced with ones in the existing CellPhoneDB dataset. For example, a gene such as Pleiotrophin (PTN) present in the single-cell data also needed to be present in the CellPhoneDB dataset. Using the single-cell transcriptomics data via slide-seq and reference data, the CellPhoneDB program defines cell types and plots them in different formats, with the two main ones being dot plots and heat map plots. The dot plot displays derived measures of the cell to cell interaction scores and p values. For the dot plot, each row shows a ligand-receptor pair, and each column shows the two interacting cell types. CellPhoneDB defines interactions and interaction levels from the gene expression level, so since the p-value is on a -log10 scale, the larger dots represent more significant interactions. By performing an interaction analysis, a significant interaction was discovered for myeloid and T-cell ligand-receptor pairs, including those between Secreted Phosphoprotein 1 (SPP1) and Fibronectin 1 (FN1), which is consistent with previous findings. It was proposed that an effective protocol would involve a filtration step where cell types would be filtered out, depending on which ligand-receptor pair is activated in that part of the tissue, as well as the incorporation of the CellPhoneDB data in a streamlined workflow pipeline. The filtration step would be in the form of a Python script that expedites the manual process necessary for dataset filtration. Being in Python allows it to be integrated with the CellPhoneDB dataset for future workflow analysis. The manual process involves filtering cell types based on what ligand/receptor pair is activated in kidney cells. One limitation of this would be the fact that some pairings are activated in multiple cells at a time, so the manual manipulation of the data is reflected prior to analysis. Using the filtration script, accurate sorting is incorporated into the CellPhoneDB database rather than waiting until the output is produced and then subsequently applying spatial data. It was envisioned that this would reveal wherein the cell various ligands and receptors are interacting with different cell types, allowing for easier identification of which cells are being impacted and why, for the purpose of disease treatment. The hope is this new computational method utilizing spatially explicit ligand-receptor association data can be used to uncover previously unknown specific interactions within kidney tissue.

Keywords: bioinformatics, Ligands, kidney tissue, receptors, spatial transcriptome

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7636 Anti-Aging Effects of Retinol and Alpha Hydroxy Acid on Elastin Fibers of Artificially Photo-Aged Human Dermal Fibroblast Cell Lines

Authors: Mohammed Jarrar, Shalini Behl, Nadia Shaheen, Abeer Fatima, Reem Nasab

Abstract:

Skin aging is a slow multifactorial process influenced by both internal as well as external factors. Ultra-violet radiations (UV), diet, smoking and personal habits are the most common environmental factors that affect skin aging. Fat contents and fibrous proteins as collagen and elastin are core internal structural components. The direct influence of UV on elastin integrity and health is crucial on aging of skin by time. The deposition of abnormal elastic material is a major marker in a photo-aged skin. Searching for compounds that may protect against cutaneous photo-damage is highly valued. Retinoids and Alpha Hydroxy Acids protective and or repairing effects of UV have been endorsed by some researchers. For consolidating a better understanding of anti and protective effects of such anti-aging agents, we evaluated the combinatory effects of various dosages of lactic acid and retinol on the dermal fibroblasts elastin levels exposed to UV. The UV exposed cells showed significant reduction in the elastin levels. A combination of drugs with a higher concentration of lactic acid (30-35 mM) and a lower concentration of retinol (10-15mg/mL) showed to work better in enhancing elastin concentration in UV exposed cells. We assume this enhancement could be the result of increased tropo-elastin gene expression stimulated by retinol and lactic acid probably repaired the UV irradiated damage by enhancing the amount and integrity of the elastin fibers.

Keywords: alpha hydroxy acid, elastin, retinol, ultraviolet radiations

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7635 Experimental Study of Tunable Layout Printed Fresnel Lens Structure Based on Dye Doped Liquid Crystal

Authors: M. Javadzadeh, H. Khoshsima

Abstract:

In this article, we present a layout printing way for producing Fresnel zone on 1294-1b doped liquid crystal with Methyl-Red azo dye. We made a Fresnel zone mask with 25 zones and radius of 5 mm using lithography technique. With layout printing way, we recorded mask’s pattern on cell with λ=532 nm solid-state diode pump laser. By recording Fresnel zone pattern on cell and making Fresnel pattern on the surface of cell, odd and even zones, will form. The printed pattern, because of Azo dye’s photoisomerization, was permanent. Experimentally, we saw focal length tunability from 32 cm to 43 cm.

Keywords: liquid crystal, lens, Fresnel zone, diffraction, Fresnel lens

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7634 Pre-Treatment of Anodic Inoculum with Nitroethane to Improve Performance of a Microbial Fuel Cell

Authors: Rajesh P.P., Md. Tabish Noori, Makarand M. Ghangrekar

Abstract:

Methanogenic substrate loss is reported to be a major bottleneck in microbial fuel cell which significantly reduces the power production capacity and coulombic efficiency (CE) of microbial fuel cell (MFC). Nitroethane is found to be a potent inhibitor of hydrogenotrophic methanogens in rumen fermentation process. Influence of nitroethane pre-treated sewage sludge inoculum on suppressing the methanogenic activity and enhancing the electrogenesis in MFC was evaluated. MFC inoculated with nitroethane pre-treated anodic inoculum demonstrated a maximum operating voltage of 541 mV, with coulombic efficiency and sustainable volumetric power density of 39.85 % and 14.63 W/m3 respectively. Linear sweep voltammetry indicated a higher electron discharge on the anode surface due to enhancement of electrogenic activity while suppressing methanogenic activity. A 63 % reduction in specific methanogenic activity was observed in anaerobic sludge pre-treated with nitroethane; emphasizing significance of this pretreatment for suppressing methanogenesis and its utility for enhancing electricity generation in MFC.

Keywords: coulombic efficiency, methanogenesis inhibition, microbial fuel cell, nitroethane

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7633 Modeling of Oligomerization of Ethylene in a Falling film Reactor for the Production of Linear Alpha Olefins

Authors: Adil A. Mohammed, Seif-Eddeen K. Fateen, Tamer S. Ahmed, Tarek M. Moustafa

Abstract:

Falling film were widely used for gas-liquid absorption and reaction process. Modeling of falling film for oligomerization of ethylene reaction to linear alpha olefins is developed. Although there are many researchers discuss modeling of falling film in many processes, there has been no publish study the simulation of falling film for the oligomerization of ethylene reaction to produce linear alpha olefins. The Comsol multiphysics software was used to simulate the mass transfer with chemical reaction in falling film absorption process. The effect of concentration profile absorption of the products through falling thickness is discussed. The effect of catalyst concentration, catalyst/co-catalyst ratio, and temperature is also studied. For the effect of the temperature, as it increase the concentration of C4 increase. For catalyst concentration and catalyst/co-catalyst ratio as they increases the concentration of C4 increases, till it reached almost constant value.

Keywords: falling film, oligomerization, comsol mutiphysics, linear alpha olefins

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7632 Preparation and Evaluation of siRNA Loaded Polymeric Nanoparticles

Authors: Riddhi Trivedi, Shrenik Shah

Abstract:

For Si RNA to be delivered various biodegradable polymers are trialed by many researchers. One of them is Chitosan (CS) nanoparticles which have been extensively studied for siRNA delivery but the stability and efficacy of such particles are highly dependent on the types of cross-linker used. Hence the attempts are made in this study with PGA To address this issue, three common cross-linkers; Ethylene glycol diacrylate (ED) and poly-D-glutamic acid (PGA) were used to prepare siRNA loaded CS-ED/PGA nanoparticles by ionic gelation method. The nanoparticles which were obtained were compared for its characterization in terms of its physicochemical properties i.e. particle size of the resultant particles, zeta potential, its encapsulation capacity in the polymer. Among all the formulations prepared with different crosslinker PGA siRNA had the smallest particle size (ranged from 120 ± 1.7 to 500 ± 10.9 nm) with zeta potential ranged from 22.1 ± 1.5 to +32.4 ± 0.5 mV, and high entrapment ( > 91%) and binding efficiencies. Similarly, CS-ED nanoparticles showed better siRNA protection during storage at 4˚C and as determined by serum protection assay. TEM micrographs revealed the assorted morphology of CS-PGA-siRNA nanoparticles in contrast to irregular morphology displayed by CS-ED-siRNA. All siRNA loaded nanoparticles were found to give initial burst release which after some time followed by a sustained release of siRNA which were loaded inside. All the formulations showed concentration-dependent cytotoxicity with when cytotoxicity performed by HeLa and normal vero cell lines.

Keywords: chitosan, siRNA, cytotoxicity, cell line study

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7631 Evaluation of Hydrocarbons in Tissues of Bivalve Mollusks from the Red Sea Coast

Authors: Asma Ahmed Aljohani, Mohammed Orif

Abstract:

The concentration of polycyclic aromatic hydrocarbons (PAH) in clam (A. glabrata) was examined in samples collected from Alseef Beach, 30 km south of Jeddah city. Gas chromatography-mass spectrometry (GC-MS) was used to analyse the 14 PAHs. The concentration of total PAHs was found to range from 11.521 to 40.149 ng/gdw with a mean concentration of 21.857 ng/gdw, which is lower compared to similar studies. The lower molecular weight PAHs with three rings comprised 18.14% of the total PAH concentrations in the clams, while the high molecular weight PAHs with four rings, five rings, and six rings account for 81.86%. Diagnostic ratios for PAH source distinction suggested pyrogenic or anthropogenic sources.

Keywords: bivalves, biomonitoring, hydrocarbons, PAHs

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7630 Evaluation of the Microscopic-Observation Drug-Susceptibility Assay Drugs Concentration for Detection of Multidrug-Resistant Tuberculosis

Authors: Anita, Sari Septiani Tangke, Rusdina Bte Ladju, Nasrum Massi

Abstract:

New diagnostic tools are urgently needed to interrupt the transmission of tuberculosis and multidrug-resistant tuberculosis. The microscopic-observation drug-susceptibility (MODS) assay is a rapid, accurate and simple liquid culture method to detect multidrug-resistant tuberculosis (MDR-TB). MODS were evaluated to determine a lower and same concentration of isoniazid and rifampin for detection of MDR-TB. Direct drug-susceptibility testing was performed with the use of the MODS assay. Drug-sensitive control strains were tested daily. The drug concentrations that used for both isoniazid and rifampin were at the same concentration: 0.16, 0.08 and 0.04μg per milliliter. We tested 56 M. tuberculosis clinical isolates and the control strains M. tuberculosis H37RV. All concentration showed same result. Of 53 M. tuberculosis clinical isolates, 14 were MDR-TB, 38 were susceptible with isoniazid and rifampin, 1 was resistant with isoniazid only. Drug-susceptibility testing was performed with the use of the proportion method using Mycobacteria Growth Indicator Tube (MGIT) system as reference. The result of MODS assay using lower concentration was significance (P<0.001) compare with the reference methods. A lower and same concentration of isoniazid and rifampin can be used to detect MDR-TB. Operational cost and application can be more efficient and easier in resource-limited environments. However, additional studies evaluating the MODS using lower and same concentration of isoniazid and rifampin must be conducted with a larger number of clinical isolates.

Keywords: isoniazid, MODS assay, MDR-TB, rifampin

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7629 Production of Hydroxy Marilone C as a Bioactive Compound from Streptomyces badius

Authors: Osama H. Elsayed, Mohsen M. S. Asker, Mahmoud A. Swelim, Ibrahim H. Abbas, Aziza I. Attwa, Mohamed E. El Awady

Abstract:

Hydroxy marilone C is a bioactive metabolite was produced from the culture broth of Streptomyces badius isolated from Egyptian soil. hydroxy marilone C was purified and fractionated by silica gel column with a gradient mobile phase dicloromethane (DCM) : Methanol then Sephadex LH-20 column using methanol as a mobile phase. It was subjected to many instruments as Infrared (IR), nuclear magnetic resonance (NMR), Mass spectroscopy (MS) and UV spectroscopy to the elucidation of its structure. It was evaluated for antioxidant, cytotoxicity against human alveolar basal epithelial cell line (A-549) and human breast adenocarcinoma cell line (MCF-7) and antiviral activities; showed that the maximum antioxidant activity was 78.8 % at 3000 µg/ml after 90 min. and the IC50 value against DPPH radical found about 1500 µg/ml after 60 min. By Using MTT assay the effect of the pure compound on the proliferation of A-549 cells and MCF-7 cells were 443 µg/ml and 147.9 µg/ml, respectively. While for detection of antiviral activity using Madin-Darby canine kidney (MDCK) cells the maximum cytotoxicity was at 27.9% and IC50 was 128.1µg/ml. The maximum concentration required for protecting 50% of the virus-infected cells against H1N1 viral cytopathogenicity (EC50) was 33.25% for 80 µg/ml. This results indicated that the hydroxy marilone C has a potential antitumor and antiviral activities.

Keywords: hydroxy marilone C, production, bioactive compound, Streptomyces badius

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7628 Modification of Fick’s First Law by Introducing the Time Delay

Authors: H. Namazi, H. T. N. Kuan

Abstract:

Fick's first law relates the diffusive flux to the concentration field, by postulating that the flux goes from regions of high concentration to regions of low concentration, with a magnitude that is proportional to the concentration gradient (spatial derivative). It is clear that the diffusion of flux cannot be instantaneous and should be some time delay in this propagation. But Fick’s first law doesn’t consider this delay which results in some errors especially when there is a considerable time delay in the process. In this paper, we introduce a time delay to Fick’s first law. By this modification, we consider that the diffusion of flux cannot be instantaneous. In order to verify this claim an application sample in fluid diffusion is discussed and the results of modified Fick’s first law, Fick’s first law and the experimental results are compared. The results of this comparison stand for the accuracy of the modified model. The modified model can be used in any application where the time delay has considerable value and neglecting its effect reflects in undesirable results.

Keywords: Fick's first law, flux, diffusion, time delay, modified Fick’s first law

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7627 Cytotoxicity of Thymoquinone Alone or in Combination with Cisplatin (CDDP) Against Oral Squamous Cell Carcinoma in Vitro

Authors: Omar M. Al Aufi, Abdulwahab Noorwali, Ahmed Al Abd, Safia Alattas, Fathya Zahran, Fahd Almutairi

Abstract:

Cisplatin (CDDP) is a potent anticancer agent used for several tumor types. Thymoquinone (TQ) is a naturally occurring compound drawing great attention as an anticancer and chemomodulator for chemotherapies. Herein, we studied the potential cytotoxicity of thymoquinone, CDDP and their combination against human oral squamous cell carcinoma cells in contrast to normal oral epithelial cells. CDDP similarly killed both head and neck squamous cell carcinoma cells (UMSCC-14C) and normal oral epithelial cells (OEC). TQ alone exerted considerable cytotoxicity against UMSCC-14C cells, while it induced a weaker killing effect against normal oral epithelial cells (OEC). The equitoxic combination of TQ and CDDP showed additive to synergistic interaction against both UMSCC-14C and OEC cells. TQ alone increased apoptotic cell fraction in UMSCC-14C cells as early as after 6 hours. In addition, prolonged exposure of UMSCC-14C to TQ alone resulted in 96.7±1.6% total apoptosis, which was increased after combination with CDDP to 99.3±1.2% in UMSCC-14C cells. On the other hand, TQ induced a marginal increase in the apoptosis in OEC and even decreased the apoptosis induced by CDDP alone. Finally, apoptosis induction results were confirmed by the change in the expression levels of p53, Bcl-2 and Caspase-9 proteins in both UMSCC-14c and OEC cells.

Keywords: thymoquinone, cisplatin, apoptosis, oral squamous cell carcinoma, P53, Caspase-9, Bcl-2

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7626 Nanoparticles Made of Amino Acid Derived Biodegradable Polymers as Promising Drug Delivery Containers

Authors: Sophio Kobauri, Tengiz Kantaria, Temur Kantaria, David Tugushi, Nina Kulikova, Ramaz Katsarava

Abstract:

Polymeric disperse systems such as nanoparticles (NPs) are of high interest for numerous applications in contemporary medicine and nanobiotechnology to a considerable potential for treatment of many human diseases. The important technological advantages of NPs usage as drug carriers (nanocontainers) are their high stability, high carrier capacity, feasibility of encapsulation of both hydrophilic or hydrophobic substances, as well as a high variety of possible administration routes, including oral application and inhalation. NPs can also be designed to allow controlled (sustained) drug release from the matrix. These properties of NPs enable improvement of drug bioavailability and might allow drug dosage decrease. The targeted and controlled administration of drugs using NPs might also help to overcome drug resistance, which is one of the major obstacles in the control of epidemics. Various degradable and non-degradable polymers of both natural and synthetic origin have been used for NPs construction. One of the most promising for the design of NPs are amino acid-based biodegradable polymers (AABBPs) which can clear from the body after the fulfillment of their function. The AABBPs are composed of naturally occurring and non-toxic building blocks such as α-amino acids, fatty diols and dicarboxylic acids. The particles designed from these polymers are expected to have an improved bioavailability along with a high biocompatibility. The present work deals with a systematic study of the preparation of NPs by cost-effective polymer deposition/solvent displacement method using AABBPs. The influence of the nature and concentration of surfactants, concentration of organic phase (polymer solution), and the ratio organic phase/inorganic(water) phase, as well as of some other factors on the size of the fabricated NPs have been studied. It was established that depending on the used conditions the NPs size could be tuned within 40-330 nm. At the next step of this research was carried out an evaluation of biocompability and bioavailability of the synthesized NPs using a stable human cell culture line – A549. It was established that the obtained NPs are not only biocompatible but they stimulate the cell growth.

Keywords: amino acids, biodegradable polymers, bioavailability, nanoparticles

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7625 Bioinformatics Approach to Identify Physicochemical and Structural Properties Associated with Successful Cell-free Protein Synthesis

Authors: Alexander A. Tokmakov

Abstract:

Cell-free protein synthesis is widely used to synthesize recombinant proteins. It allows genome-scale expression of various polypeptides under strictly controlled uniform conditions. However, only a minor fraction of all proteins can be successfully expressed in the systems of protein synthesis that are currently used. The factors determining expression success are poorly understood. At present, the vast volume of data is accumulated in cell-free expression databases. It makes possible comprehensive bioinformatics analysis and identification of multiple features associated with successful cell-free expression. Here, we describe an approach aimed at identification of multiple physicochemical and structural properties of amino acid sequences associated with protein solubility and aggregation and highlight major correlations obtained using this approach. The developed method includes: categorical assessment of the protein expression data, calculation and prediction of multiple properties of expressed amino acid sequences, correlation of the individual properties with the expression scores, and evaluation of statistical significance of the observed correlations. Using this approach, we revealed a number of statistically significant correlations between calculated and predicted features of protein sequences and their amenability to cell-free expression. It was found that some of the features, such as protein pI, hydrophobicity, presence of signal sequences, etc., are mostly related to protein solubility, whereas the others, such as protein length, number of disulfide bonds, content of secondary structure, etc., affect mainly the expression propensity. We also demonstrated that amenability of polypeptide sequences to cell-free expression correlates with the presence of multiple sites of post-translational modifications. The correlations revealed in this study provide a plethora of important insights into protein folding and rationalization of protein production. The developed bioinformatics approach can be of practical use for predicting expression success and optimizing cell-free protein synthesis.

Keywords: bioinformatics analysis, cell-free protein synthesis, expression success, optimization, recombinant proteins

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7624 Synergistic Effect of Curcumin and Insulin on GLUT4 Translocation in C2C12 Cell

Authors: Javad Mohiti-Ardekani, Shabodin Asadii, Ali Moradi

Abstract:

Introduction: Curcumin, the yellow pigment in turmeric, has been shown as an anti-diabetic agent for centuries but only in recent few years, its mechanism of action has been under investigation. Some studies showed that curcumin might exert its anti-diabetic effect via increasing glucose transporter isotype-4 (GLUT4) gene and glycoprotein contents in cells. To investigate this possibility, we investigate the effects of extract and commercial curcumin with and without insulin on GLUT4 translocation from intracellular compartments of nuclear or endoplasmic reticulum membranes (N/ER) into the cytoplasmic membrane (CM). Methods and Material: C2C12 myoblastic cell line were seeded in DMEM plus 20 % FBS and differentiated to myotubes using 2 % horse serum. After myotubes formation, 40 µmolar Extract and Commercial curcumin, with or without insulin as intervention, and as control 1 % DMSO were added for 3 h. Cells were washed and homogenized followed by ultracentrifuge fractionation, protein separation by SDS-PAGE and GLUT4 detection using semi-quantitative Western blotting. Data analysis was done by two independent samples t-test for comparison of mean ± SD of GLUT4 percent in categories. GLUT4 contents were higher in CM groups curcumin and curcumin with insulin in comparison to 1 % DMSO-treated myotubes control group. Results: As our results have shown extract and commercial curcumin induces GLUT4 translocation from intra-cell into cell surface. The results have also shown synergic effect of curcumin on translocation of GLUT4 from intra-cell into cell surface in the presence of 100 nm insulin. Discussion: We conclude that curcumin may be a choice of type-2 diabetes mellitus treatment because its extract and commercial enhances GLUT4 contents in CM where it facilitates glucose entrance into the cell. However, it is necessary to trace the signaling pathways which are activated by curcumin.

Keywords: Curcumin, insulin, Diabetes type-2, GLUT4

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7623 Magneto-Luminescent Biocompatible Complexes Based on Alloyed Quantum Dots and Superparamagnetic Iron Oxide Nanoparticles

Authors: A. Matiushkina, A. Bazhenova, I. Litvinov, E. Kornilova, A. Dubavik, A. Orlova

Abstract:

Magnetic-luminescent complexes based on superparamagnetic iron oxide nanoparticles (SPIONs) and semiconductor quantum dots (QDs) have been recognized as a new class of materials that have high potential in modern medicine. These materials can serve for theranostics of oncological diseases, and also as a target agent for drug delivery. They combine the qualities characteristic of magnetic nanoparticles, that is, magneto-controllability and the ability to local heating under the influence of an external magnetic field, as well as phosphors, due to luminescence of which, for example, early tumor imaging is possible. The complexity of creating complexes is the energy transfer between particles, which quenches the luminescence of QDs in complexes with SPIONs. In this regard, a relatively new type of alloyed (CdₓZn₁₋ₓSeᵧS₁₋ᵧ)-ZnS QDs is used in our work. The presence of a sufficiently thick gradient semiconductor shell in alloyed QDs makes it possible to reduce the probability of energy transfer from QDs to SPIONs in complexes. At the same time, Forster Resonance Energy Transfer (FRET) is a perfect instrument to confirm the formation of complexes based on QDs and different-type energy acceptors. The formation of complexes in the aprotic bipolar solvent dimethyl sulfoxide is ensured by the coordination of the carboxyl group of the stabilizing QD molecule (L-cysteine) on the surface iron atoms of the SPIONs. An analysis of the photoluminescence (PL) spectra has shown that a sequential increase in the SPIONs concentration in the samples is accompanied by effective quenching of the luminescence of QDs. However, it has not confirmed the formation of complexes yet, because of a decrease in the PL intensity of QDs due to reabsorption of light by SPIONs. Therefore, a study of the PL kinetics of QDs at different SPIONs concentrations was made, which demonstrates that an increase in the SPIONs concentration is accompanied by a symbatic reduction in all characteristic PL decay times. It confirms the FRET from QDs to SPIONs, which indicates the QDs/SPIONs complex formation, rather than a spontaneous aggregation of QDs, which is usually accompanied by a sharp increase in the percentage of the QD fraction with the shortest characteristic PL decay time. The complexes have been studied by the magnetic circular dichroism (MCD) spectroscopy that allows one to estimate the response of magnetic material to the applied magnetic field and also can be useful to check SPIONs aggregation. An analysis of the MCD spectra has shown that the complexes have zero residual magnetization, which is an important factor for using in biomedical applications, and don't contain SPIONs aggregates. Cell penetration, biocompatibility, and stability of QDs/SPIONs complexes in cancer cells have been studied using HeLa cell line. We have found that the complexes penetrate in HeLa cell and don't demonstrate cytotoxic effect up to 25 nM concentration. Our results clearly demonstrate that alloyed (CdₓZn₁₋ₓSeᵧS₁₋ᵧ)-ZnS QDs can be successfully used in complexes with SPIONs reached new hybrid nanostructures, which combine bright luminescence for tumor imaging and magnetic properties for targeted drug delivery and magnetic hyperthermia of tumors. Acknowledgements: This work was supported by the Ministry of Science and Higher Education of Russian Federation, goszadanie no. 2019-1080 and was financially supported by Government of Russian Federation, Grant 08-08.

Keywords: alloyed quantum dots, magnetic circular dichroism, magneto-luminescent complexes, superparamagnetic iron oxide nanoparticles

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7622 Study of Corrosion in Structures due to Chloride Infiltration

Authors: Sukrit Ghorai, Akku Aby Mathews

Abstract:

Corrosion in reinforcing steel is the leading cause for deterioration in concrete structures. It is an electrochemical process which leads to volumetric change in concrete and causes cracking, delamination and spalling. The objective of the study is to provide a rational method to estimate the probable chloride concentration at the reinforcement level for a known surface chloride concentration. The paper derives the formulation of design charts to aid engineers for quick calculation of the chloride concentration. Furthermore, the paper focuses on comparison of durability design against corrosion with American, European and Indian design standards.

Keywords: chloride infiltration, concrete, corrosion, design charts

Procedia PDF Downloads 412
7621 Optimization of Parameters for Electrospinning of Pan Nanofibers by Taguchi Method

Authors: Gamze Karanfil Celep, Kevser Dincer

Abstract:

The effects of polymer concentration and electrospinning process parameters on the average diameters of electrospun polyacrylonitrile (PAN) nanofibers were experimentally investigated. Besides, mechanical and thermal properties of PAN nanofibers were examined by tensile test and thermogravimetric analysis (TGA), respectively. For this purpose, the polymer concentration, solution feed rate, supply voltage and tip-to-collector distance were determined as the control factors. To succeed these aims, Taguchi’s L16 orthogonal design (4 parameters, 4 level) was employed for the experimental design. Optimal electrospinning conditions were defined using the signal-to-noise (S/N) ratio that was calculated from diameters of the electrospun PAN nanofibers according to "the-smaller-the-better" approachment. In addition, analysis of variance (ANOVA) was evaluated to conclude the statistical significance of the process parameters. The smallest diameter of PAN nanofibers was observed. According to the S/N ratio response results, the most effective parameter on finding out of nanofiber diameter was determined. Finally, the Taguchi design of experiments method has been found to be an effective method to statistically optimize the critical electrospinning parameters used in nanofiber production. After determining the optimum process parameters of nanofiber production, electrical conductivity and fuel cell performance of electrospun PAN nanofibers on the carbon papers will be evaluated.

Keywords: nanofiber, electrospinning, polyacrylonitrile, Taguchi method

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7620 Non-Signaling Chemokine Receptor CCRL1 and Its Active Counterpart CCR7 in Prostate Cancer

Authors: Yiding Qu, Svetlana V. Komarova

Abstract:

Chemokines acting through their cognate chemokine receptors guide the directional migration of the cell along the chemokine gradient. Several chemokine receptors were recently identified as non-signaling (decoy), based on their ability to bind the chemokine but produce no measurable signal in the cell. The function of these decoy receptors is not well understood. We examined the expression of a decoy receptor CCRL1 and a signaling receptor that binds to the same ligands, CCR7, in prostate cancer using publically available microarray data (www.oncomine.org). The expression of both CCRL1 and CCR7 increased in an approximately half of prostate carcinoma samples and the majority of metastatic cancer samples compared to normal prostate. Moreover, the expression of CCRL1 positively correlated with the expression of CCR7. These data suggest that CCR7 and CCRL1 can be used as clinical markers for the early detection of transformation from carcinoma to metastatic cancer. In addition, these data support our hypothesis that the non-signaling chemokine receptors actively stimulate cell migration.

Keywords: bioinformatics, cell migration, decoy receptor, meta-analysis, prostate cancer

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7619 Cellular Energy Metabolism Decreases with Age in the Trophocytes and Oenocytes of Honeybees (Apis Mellifera)

Authors: Chin-Yuan Hsu, Yu-Lung Chuang

Abstract:

The expression, concentration, and activity of mitochondrial energy-utilized molecules and cellular energy-regulated molecules decreased with age in the trophocytes and oenocytes of honeybees (Apis mellifera), but those of cellular energy-metabolized molecules is unknown. In this study, the expression, concentration, and activity of cellular energy-metabolized molecules were assayed in the trophocytes and fat cells of young and old worker bees by using the techniques of cell and biochemistry. The results showed that (i) the •-hydroxylacyl-coenzyme A dehydrogenase (HOAD) activity/citrate synthase (CS) activity ratio, non-esterified fatty acids concentrations, the expression of eukaryotic initiation factor 4E, and the expression of phosphorylated eIF4E binding protein 1 decreased with age; (ii) fat and glycogen accumulation increased with age; and (iii) the pyruvate dehydrogenase (PDH) activity/citrate synthase (CS) activity ratio was not correlated with age. These finding indicated that •-oxidation (HOAD/CS) and protein synthsis decreased with age. Glycolysis (PDH/CS) was unchanged with age. The most likely reason is that sugars are the vital food of worker bees. Taken together these data reveal that young workers have higher cellular energy metabolism than old workers and that aging results in a decline in the cellular energy metabolism in worker honeybees.

Keywords: aging, energy, honeybee, metabolism

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7618 Developing Scaffolds for Tissue Regeneration using Low Temperature Plasma (LTP)

Authors: Komal Vig

Abstract:

Cardiovascular disease (CVD)-related deaths occur in 17.3 million people globally each year, accounting for 30% of all deaths worldwide, with a predicted annual incidence of deaths to reach 23.3 million globally by 2030. Autologous bypass grafts remain an important therapeutic option for the treatment of CVD, but the poor quality of the donor patient’s blood vessels, the invasiveness of the resection surgery, and postoperative movement restrictions create issues. The present study is aimed to improve the endothelialization of intimal surface of graft by using low temperature plasma (LTP) to increase the cell attachment and proliferation. Polytetrafluoroethylene (PTFE) was treated with LTP. Air was used as the feed-gas, and the pressure in the plasma chamber was kept at 800 mTorr. Scaffolds were also modified with gelatin and collagen by dipping method. Human umbilical vein endothelial cells (HUVEC) were plated on the developed scaffolds, and cell proliferation was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by microscopy. mRNA expressions levels of different cell markers were investigated using quantitative real-time PCR (qPCR). XPS confirmed the introduction of oxygenated functionalities from LTP. HUVEC cells showed 80% seeding efficiency on the scaffold. Microscopic and MTT assays indicated increase in cell viability in LTP treated scaffolds, especially when treated with gelatin or collagen, compared to untreated scaffolds. Gene expression studies shows enhanced expression of cell adhesion marker Integrin- α 5 gene after LTP treatment. LTP treated scaffolds exhibited better cell proliferation and viability compared to untreated scaffolds. Protein treatment of scaffold increased cell proliferation. Based on our initial results, more scaffolds alternatives will be developed and investigated for cell growth and vascularization studies. Acknowledgments: This work is supported by the NSF EPSCoR RII-Track-1 Cooperative Agreement OIA-2148653.

Keywords: LTP, HUVEC cells, vascular graft, endothelialization

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7617 Sirt1 Activators Promote Skin Cell Regeneration and Cutaneous Wound Healing

Authors: Hussain Mustatab Wahedi, Sun You Kim

Abstract:

Skin acts as a barrier against the harmful environmental factors. Integrity and timely recovery of the skin from injuries and harmful effects of radiations is thus very important. This study aimed to investigate the importance of Sirt1 in the recovery of skin from UVB-induced damage and cutaneous wounds by using natural and synthetic novel Sirt1 activators. Juglone, known as a natural Pin1 inhibitor, and NED416 a novel synthetic Sirt1 activator were checked for their ability to regulate the expression and activity of Sirt1 and hence photo-damage and wound healing in cultured skin cells (NHDF and HaCaT cells) and mouse model by using Sirt1 siRNA knockdown, cell migration assay, GST-Pulldown assay, western blot analysis, tube formation assay, and immunohistochemistry. Interestingly, Sirt1 knockdown inhibited skin cell migration in vitro. Juglone up regulated the expression of Sirt1 in both the cell lines under normal and UVB irradiated conditions, enhanced Sirt1 activity and increased the cell viability by reducing reactive oxygen species synthesis and apoptosis. Juglone promoted wound healing by increasing cell migration and angiogenesis through Cdc42/Rac1/PAK, MAPKs and Smad pathways in skin cells. NED416 upregulated Sirt1 expression in HaCaT and NHDF cells as well as increased Sirt1 activity. NED416 promoted the process of wound healing in early as well as later stages by increasing macrophage recruitment, skin cell migration, and angiogenesis through Cdc42/Rac1 and MAPKs pathways. So, both these compounds activated Sirt1 and promoted the process of wound healing thus pointing towards the possible role of Sirt1 in skin regeneration and wound healing.

Keywords: skin regeneration, wound healing, Sirt1, UVB light

Procedia PDF Downloads 189
7616 Optimum Design for Cathode Microstructure of Solid Oxide Fuel Cell

Authors: M. Riazat, H. Abdolvand, M. Baniassadi

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In this present work, 3D reconstruction of cathode of SOFC is developed with various volume fractions and porosity. Three Phase Boundary (TPB) of construction of such derived micro structures is calculated. The neural network is used to optimize the porosity and volume fraction of each phase to reach a structure with maximum TPB.

Keywords: fuel cell, solid oxide, TPB, 3D reconstruction

Procedia PDF Downloads 325
7615 SOCS1 Inhibits MDR1 in Mammary Cell Carcinoma Reverses Multidrug Resistance

Authors: Debasish Pradhan, Shaktiprasad Pradhan, Rakesh Kumar Pradhan, Gitanjali Tripathy

Abstract:

Suppressors of cytokine signalling (SOCS1), a newly indentified antiapoptotic molecule is a downstream effector of the receptor tyrosine kinase-Ras signalling pathway. The current study has uncovered that SOCS1 may have wide and imperative capacities, particularly because of its close correlation with malignant tumors. To investigate the impact of SOCS1 on MDR, we analyzed the expression of P-gp and SOCS1 by immunohistochemistry and found there was a positive correlation between them. At that point, we effectively interfered with RNA translation by the contamination of siRNA of SOCS1 into MCF7/ADM breast cancer cell lines through a lentivirus, and the expression of the target gene was significantly inhibited. After RNAi, the drug resistance was reduced altogether and the expression of MDR1 mRNA and P-gp in MCF7/ADM cell lines demonstrated a significant decrease. Likewise, the expression of P53 protein increased in a statistically significant manner (p ≤ 0.01) after RNAi exposure. Moreover, flow cytometry analysis uncovers that cell cycle and anti-apoptotic enhancing capacity of cells changed after RNAi treatment. These outcomes proposed SOCS1 may take part in breast cancer MDR by managing MDR1 and P53 expression, changing cell cycle and enhancing the anti-apoptotic ability.

Keywords: breast cancer, multidrug resistance, SOCS1 gene, MDR1 gene, RNA interference

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7614 Feasibility of Leukemia Cancer Treatment (K562) by Atmospheric Pressure Plasma Jet

Authors: Mashayekh Amir Shahriar, Akhlaghi Morteza, Rajaee Hajar, Khani Mohammad Reza, Shokri Babak

Abstract:

A new and novel approach in medicine is the use of cold plasma for various applications such as sterilization blood coagulation and cancer cell treatment. In this paper a pin-to-hole plasma jet suitable for biological applications is investigated, characterized and the possibility and feasibility of cancer cell treatment is evaluated. The characterization includes power consumption via Lissajous method, thermal behavior of plasma using Infra-red camera as a novel method, Optical Emission Spectroscopy (OES) to determine the species that are generated. Treatment of leukemia cancer cells is also implemented and MTT assay is used to evaluate viability.

Keywords: Atmospheric Pressure Plasma Jet (APPJ), Plasma Medicine, Cancer cell treatment, leukemia, Optical Emission

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7613 PBI Based Composite Membrane for High Temperature Polymer Electrolyte Membrane Fuel Cells

Authors: Kwangwon Seo, Haksoo Han

Abstract:

Al-Si was synthesized and introduced in poly 2,2’-m-(phenylene)-5,5’-bibenzimidazole (PBI). As a result, a series of five Al-Si/PBI composite (ASPBI) membranes (0, 3, 6, 9, and 12 wt.%) were developed and characterized for application in high temperature polymer electrolyte membrane fuel cells (HT-PEMFCs). The chemical and morphological structure of ASPBI membranes were analyzed by Fourier transform infrared spectroscopy, X-ray diffractometer and scanning electron microscopy. According to the doping level test and thermogravimetric analysis, as the concentration of Al-Si increased, the doping level increased up to 475%. Moreover, the proton conductivity, current density at 0.6V, and maximum power density of ASPBI membranes increased up to 0.31 Scm-1, 0.320 Acm-2, and 0.370 Wcm-2, respectively, because the increased concentration of Al-Si allows the membranes to hold more PA. Alternatively, as the amount of Al-Si increased, the tensile strength of PA-doped and -undoped membranes decreased. This was resulted by both excess PA and aggregation, which can cause serious degradation of the membrane and induce cracks. Moreover, the PA-doped and -undoped ASPBI12 had the lowest tensile strength. The improved performances of ASPBI membranes imply that ASPBI membranes are possible candidates for HT-PEMFC applications. However, further studies searching to improve the compatibility between PBI matrix and inorganic and optimize the loading of Al-Si should be performed.

Keywords: composite membrane, high temperature polymer electrolyte membrane fuel cell, membrane electrode assembly, polybenzimidazole, polymer electrolyte membrane, proton conductivity

Procedia PDF Downloads 529