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    Creating Random Mutations on The Human Transglutaminase 2 Gene and Lysozyme Gene to Increase The Solubility of the Enzymes in order to Perform Promiscuous Organic Reactions Using The Recombinant Enzymes

    , M.Sc. Thesis Sharif University of Technology Moghaddasi, Ahmad (Author) ; Kalhor, Hamid Reza (Supervisor)
    Abstract
    Transglutaminase 2 enzyme is expressed in various parts of the human body and is present in almost all tissue cells. The function of this enzyme is multiple and it catalyzes different reactions depending on where it is located in the cell. The most important activities of this enzyme are: isopeptide bond formation between the side chains of two proteins (transamidation reaction), diamidation reaction (hydrolysis of amide bond), esterification, kinase activity. The hen egg white lysozyme protein is another natural antimicrobial protein that has been extensively studied. The muramidase activity of egg white lysozyme against highly gram-positive bacteria is well known, and this property has... 

    PCR Amplification Prediction using Machine Learning

    , M.Sc. Thesis Sharif University of Technology Latifian, Niloofar (Author) ; Hossein Khalaj, Babak (Supervisor)
    Abstract
    Polymerase Chain Reaction (PCR) is a laboratory method for amplifying a part of DNA. This method is used in determining the sequence of genes, detecting pathogenic agents in epidemics, creating genetic changes in bacteria, diseases, plants and even animals. Many factors affect the quality of the reaction. Each of these factors can be effective in amplifying the target in DNA. If we can predict the result of PCR using the factors involved in the reaction, it will save a lot of money and time. The aim of this research is to predict the result of PCR amplification using machine learning methods. For this purpose, two methods are proposed: feature-based method and string-based method. In the... 

    Design of a Microfluidic Digital Droplet PCR

    , M.Sc. Thesis Sharif University of Technology Abedini, Ali (Author) ; Saeedi, Mohammad Saeed (Supervisor)
    Abstract
    Polymerase chain reaction, abbreviated as PCR, is a method of amplifying the number of copies of a desired DNA sequence through special protocols. The rapid advance of microfluidic devices and the emerging concept of digital microfluidics has resulted into the development of digital droplet PCR (ddPCR) systems with their significant uses in the detection of rare mutations, cancer diagnosis, and surveillance. A large number of micron-sized droplets are required to perform ddPCR. in this study, To investigate the gradient of confinements induced droplet self-breakup mechanism, we carried out the computational fluid dynamics (CFD) simulations for the two-phase flow using a commercial software... 

    Cloning and Production of the Flavin Reductase Enzyme in Order to Perform Organic Reaction and Synthesis of Cation Exchange Column for Protein Purification

    , M.Sc. Thesis Sharif University of Technology Hayati, Fatemeh (Author) ; Kalhor, Hamid Reza (Supervisor)
    Abstract
    The reductase enzymes perform an important function in the cell. These enzymes are usually coupled with other enzymes and participate in reactions such as halogenation, reduction of alkene, and carboxylic acids. These reactions are carried out by reproducing cofactors such as FADH2 and NAD+; however, there is no evidence that this enzyme alone can perform an organic reaction. In this work, we aim to examine whether recombinant flavin reductase can carry out organic reactions. In order to amplify the flavin reductase enzyme gene from the E. coli genome by PCR (Polymerase Chain Reaction) method, two synthetic primers were designed; The forward primer of the gene contained a cleavage site for... 

    Design and Fabrication of a Microfluidic Digital PCR

    , M.Sc. Thesis Sharif University of Technology Bahrami, Mohammad Amin (Author) ; Saeedi, Mohammad Saeed (Supervisor) ; Firouzabadi, Bahar (Supervisor)
    Abstract
    Polymerase chain reaction, abbreviated as PCR, is a method of amplifying the number of copies of a desired DNA sequence through special protocols. The rapid advance of microfluidic devices and the emerging concept of digital microfluidics has resulted into the development of digital droplet PCR (ddPCR) systems with their significant uses in the detection of rare mutations, cancer diagnosis, and surveillance. In this study, an integrated ddPCR microfluidic chip alongside its thermoelectric thermal cycler and optical analysis systems have been designed, fabricated and their performance has been tested and evaluated. It was shown that the effect of master mold’s surface quality, the material of... 

    Study, Optimization and Construction of a Microfluidic Gene Amplification Device by Using Thin Film Layer Method

    , M.Sc. Thesis Sharif University of Technology Eslami, Sara Sadat (Author) ; Vosoughi, Manouchehr (Supervisor) ; Alemzadeh, Iran (Co-Supervisor) ; Shamloo, Amir (Co-Supervisor)
    Abstract
    Polymerase Chain Reaction (PCR) is a process in which a special piece of a gene is amplified millions of times over a short period. This method has been of paramount importance in different fields of research and has been applied for different applications. PCR requires thermal cycling, or repeated temperature changes between two or three discrete temperatures to amplify specific nucleic acid target sequences. To achieve such thermal cycling, conventional bench-top thermal cyclers generally use a metal heating block powered by Peltier elements or benefit from forced convection heat transfer. Due to the fact that these methods are time consuming, it seems that design and fabrication of a fast... 

    Design, Simulation and Construction of a Rapid Gene Amplification Microfluidic Device Using Polymerase Chain Reaction (PCR) Method

    , M.Sc. Thesis Sharif University of Technology Amadeh, Ali (Author) ; Shamloo, Amir (Supervisor)
    Abstract
    Polymerase chain reaction method is a conventional method for obtaining multiple copies of a specified segment in the DNA molecule and to amplify the DNA molecule itself. Therefore, this method has been of paramount importance in different fields of research and has been applied for different applications. PCR requires thermal cycling, or repeated temperature changes between two or three discrete temperatures to amplify specific nucleic acid target sequences. To achieve such thermal cycling, conventional bench-top thermal cyclers generally use a metal heating block powered by Peltier elements or benefit from forced convection heat transfer. Although efforts have been made in order to reduce... 

    Expression of functional recombinant human tissue transglutaminase (TG2) using the bac-to-bac baculovirus expression system

    , Article Advanced Pharmaceutical Bulletin ; Volume 6, Issue 1 , 2016 , Pages 49-56 ; 22285881 (ISSN) Yazdani, Y ; Azari, S ; Kalhor, H. R ; Sharif University of Technology
    Tabriz University of Medical Sciences 
    Abstract
    Tissue transglutaminase (TG2) is a unique multifunctional enzyme. The enzyme possesses enzymatic activities such as transamidation/crosslinking and non-enzymatic functions such as cell migration and signal transduction. TG2 has been shown to be involved in molecular mechanisms of cancers and several neurodegenerative diseases such as Alzheimer's disease. The present study aimed at cloning and expression of full length human TG2 in Bac-to-Bac baculovirus expression system and evaluation of its activity. Methods: pFastBac HTA donor vector containing coding sequence of human TG2 was constructed. The construct was transformed to DH10Bac for generating recombinant bacmid. The verified bacmid was... 

    Bottom-up synthesis of nitrogen and oxygen co-decorated carbon quantum dots with enhanced DNA plasmid expression

    , Article Colloids and Surfaces B: Biointerfaces ; Volume 184 , 2019 ; 09277765 (ISSN) Yadegari, A ; Khezri, J ; Esfandiari, S ; Mahdavi, H ; Karkhane, A. A ; Rahighi, R ; Heidarimoghadam, R ; Tayebi, L ; Hashemi, E ; Farmany, A ; Sharif University of Technology
    Elsevier B.V  2019
    Abstract
    In this paper, a bottom-up hydrothermal route is reported for the synthesis of oxygen and nitrogen co-decorated carbon quantum dots (CQDs) using ammonium hydrogen citrate (AHC) as a single precursor. DLS data approved the formation of 4.0 nm (average size) CQDs. XRD pattern shows the interlayer spacing (002) of 3.5 Å for CQDs, which is exactly the same as that of crystalline graphite. XPS and FTIR spectra verified the formation of oxygen and nitrogen functional groups on the CQDs surface. Co-decorated carboxyl, hydroxyl and amine groups on the CQDs surfaces make them as promising polyelectrolyte for gene delivery. Toxicity assay showed a survival rate of 70% under different incubation times... 

    MicroRNA profiling reveals important functions of miR-125b and let-7a during human retinal pigment epithelial cell differentiation

    , Article Experimental Eye Research ; Volume 190 , 2020 Shahriari, F ; Satarian, L ; Moradi, S ; Sharifi Zarchi, A ; Günther, S ; Kamal, A ; Totonchi, M ; Mowla, S. J ; Braun, T ; Baharvand, H ; Sharif University of Technology
    Academic Press  2020
    Abstract
    Retinal pigment epithelial (RPE) cells are indispensable for eye organogenesis and vision. To realize the therapeutic potential of in vitro-generated RPE cells for cell-replacement therapy of RPE-related retinopathies, molecular mechanisms of RPE specification and maturation need to be investigated. So far, many attempts have been made to decipher the regulatory networks involved in the differentiation of human pluripotent stem cells into RPE cells. Here, we exploited a highly-efficient RPE differentiation protocol to determine global expression patterns of microRNAs (miRNAs) during human embryonic stem cell (hESC) differentiation into RPE using small RNA sequencing. Our results revealed a... 

    Expression and function of c1orf132 long-noncoding rna in breast cancer cell lines and tissues

    , Article International Journal of Molecular Sciences ; Volume 22, Issue 13 , 2021 ; 16616596 (ISSN) Shafaroudi, A. M ; Sharifi Zarchi, A ; Rahmani, S ; Nafissi, N ; Mowla, S. J ; Lauria, A ; Oliviero, S ; Matin, M. M ; Sharif University of Technology
    MDPI  2021
    Abstract
    miR-29b2 and miR-29c play a suppressive role in breast cancer progression. C1orf132 (also named MIR29B2CHG) is the host gene for generating both microRNAs. However, the region also expresses longer transcripts with unknown functions. We employed bioinformatics and experimental approaches to decipher C1orf132 expression and function in breast cancer tissues. We also used the CRISPR/Cas9 technique to excise a predicted C1orf132 distal promoter and followed the behavior of the edited cells by real-time PCR, flow cytometry, migration assay, and RNA-seq techniques. We observed that C1orf132 long transcript is significantly downregulated in triple-negative breast cancer. We also identified a... 

    Laboratory detection methods for the human coronaviruses

    , Article European Journal of Clinical Microbiology and Infectious Diseases ; Volume 40, Issue 2 , 2021 , Pages 225-246 ; 09349723 (ISSN) Shabani, E ; Dowlatshahi, S ; Abdekhodaie, M. J ; Sharif University of Technology
    Springer Science and Business Media Deutschland GmbH  2021
    Abstract
    Coronaviruses are a group of envelop viruses which lead to diseases in birds and mammals as well as human. Seven coronaviruses have been discovered in humans that can cause mild to lethal respiratory tract infections. HCoV-229E, HCoV-OC43, HCoV-NL63, and HCoV-HKU1 are the low-risk members of this family and the reason for some common colds. Besides, SARS-CoV, MERS-CoV, and newly identified SARS-CoV-2, which is also known as 2019-nCoV, are the more dangerous viruses. Due to the rapid spread of this novel coronavirus and its related disease, COVID-19, a reliable, simple, fast, and low-cost detection method is necessary for patient diagnosis and tracking worldwide. Human coronaviruses detection... 

    Gold nanoparticles loaded into niosomes: A novel approach for enhanced antitumor activity against human ovarian cancer

    , Article Advanced Powder Technology ; Volume 32, Issue 12 , 2021 , Pages 4711-4722 ; 09218831 (ISSN) Rezaie Amale, F ; Ferdowsian, S ; Hajrasouliha, S ; Kazempoor, R ; Mirzaie, A ; Sedigh Dakkali, M ; Akbarzadeh, I ; Mohammadmahdi Meybodi, S ; Mirghafouri, M ; Sharif University of Technology
    Elsevier B.V  2021
    Abstract
    In the current study, gold nanoparticles (AuNPs) were prepared using the green synthesis method using Artemisia annua extract, loaded into niosomes, and investigated their cytotoxicity and apoptotic effects. To optimize the niosomal formulation containing AuNPs, the effects of surfactants: cholesterol molar ratio, Span 60: Tween 60 M ratio, and AuNP concentration (µg/mL) were investigated. After examining the drug release profile, mathematical models were assessed to predict release kinetic. The cytotoxicity of noisome encapsulated AuNPs and free AuNPs was evaluated against human ovarian cancer cell line (A2780) by MTT assay. The apoptotic/necrosis ratio was studied using flow cytometry as... 

    Dna-Rna hybrid (R-loop): From a unified picture of the mammalian telomere to the genome-wide profile

    , Article Cells ; Volume 10, Issue 6 , 2021 ; 20734409 (ISSN) Rassoulzadegan, M ; Sharifi Zarchi, A ; Kianmehr, L ; Sharif University of Technology
    MDPI  2021
    Abstract
    Local three-stranded DNA/RNA hybrid regions of genomes (R-loops) have been detected either by binding of a monoclonal antibody (DRIP assay) or by enzymatic recognition by RNaseH. Such a structure has been postulated for mouse and human telomeres, clearly suggested by the identification of the complementary RNA Telomeric repeat-containing RNA “TERRA”. However, the tremendous disparity in the information obtained with antibody-based technology drove us to investigate a new strategy. Based on the observation that DNA/RNA hybrids in a triplex complex genome co-purify with the double-stranded chromosomal DNA fraction, we developed a direct preparative approach from total protein-free cellular... 

    Natural compounds for skin tissue engineering by electrospinning of nylon-Beta vulgaris

    , Article ASAIO Journal ; Volume 64, Issue 2 , 2018 , Pages 261-269 ; 10582916 (ISSN) Ranjbarvan, P ; Mahmoudifard, M ; Kehtari, M ; Babaie, A ; Hamedi, S ; Mirzaei, S ; Soleimani, M ; Hosseinzadeh, S ; Sharif University of Technology
    Lippincott Williams and Wilkins  2018
    Abstract
    Natural compounds containing polysaccharide ingredients have been employed as candidates for treatment of skin tissue. Herein, for the first time, electrospinning setup was proposed to fabricate an efficient composite nanofibrous structure of Beta vulgaris (obtained from Beet [Chenopodiaceae or Amaranthaceae]) belonged to polysaccharides and an elastic polymer named nylon 66 for skin tissue engineering. Both prepared scaffolds including noncomposite and composite types were studied by Scanning electron microscope (SEM), Fourier transform infrared (FTIR) spectroscopy, mechanical assay, and contact angle. Scanning electron microscope examinations have approved the uniform and homogeneous... 

    An integrated analysis to predict micro-RNAs targeting both stemness and metastasis in breast cancer stem cells

    , Article Journal of Cellular and Molecular Medicine ; Volume 23, Issue 4 , 2019 , Pages 2442-2456 ; 15821838 (ISSN) Rahimi, M ; Sharifi Zarchi, A ; Firouzi, J ; Azimi, M ; Zarghami, N ; Alizadeh, E ; Ebrahimi, M ; Sharif University of Technology
    Blackwell Publishing Inc  2019
    Abstract
    Several evidences support the idea that a small population of tumour cells representing self-renewal potential are involved in initiation, maintenance, metastasis, and outcomes of cancer therapy. Elucidation of microRNAs/genes regulatory networks activated in cancer stem cells (CSCs) is necessary for the identification of new targets for cancer therapy. The aim of the present study was to predict the miRNAs pattern, which can target both metastasis and self-renewal pathways using integration of literature and data mining. For this purpose, mammospheres derived from MCF-7, MDA-MB231, and MDA-MB468 were used as breast CSCs model. They had higher migration, invasion, and colony formation... 

    Oral administration of lithium chloride ameliorate spinal cord injury-induced hyperalgesia in male rats

    , Article PharmaNutrition ; Volume 21 , 2022 ; 22134344 (ISSN) Rahimi, G ; Mirsadeghi, S ; Rahmani, S ; Izadi, A ; Ghodsi, Z ; Ghodsi, S. M ; Rahimi Movaghar, V ; Kiani, S ; Sharif University of Technology
    Elsevier B.V  2022
    Abstract
    Background: Numerous studies have described the neuroprotective effect of lithium in spinal cord injury in addition to its ameliorative impact on pain sensation. In the present study, we aim to examine the efficacy of 85 mg/kg as well as 50 mg/kg dosage of the lithium chloride (LiCl) through oral consumption in spinal cord injured rats and their effect on gene expression of three candidate genes, corresponding to the hyper-sensitization. Methods: Adult Wistar (male) rats were divided into four experimental groups: control; oral administration of LiCl with 85 mg/kg and 50 mg/kg dosage; and 10 % sucrose receiver as the vehicle. BBB and heat plantar tests were performed weekly throughout four... 

    Point-of-use rapid detection of sars-cov-2: Nanotechnology-enabled solutions for the covid-19 pandemic

    , Article International Journal of Molecular Sciences ; Volume 21, Issue 14 , 2020 , Pages 1-23 Rabiee, N ; Bagherzadeh, M ; Ghasemi, A ; Zare, H ; Ahmadi, S ; Fatahi, Y ; Dinarvand, R ; Rabiee, M ; Ramakrishna, S ; Shokouhimehr, M ; Varma, R. S ; Sharif University of Technology
    MDPI AG  2020
    Abstract
    Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused the COVID-19 pandemic that has been spreading around the world since December 2019. More than 10 million affected cases and more than half a million deaths have been reported so far, while no vaccine is yet available as a treatment. Considering the global healthcare urgency, several techniques, including whole genome sequencing and computed tomography imaging have been employed for diagnosing infected people. Considerable efforts are also directed at detecting and preventing different modes of community transmission. Among them is the rapid detection of virus presence on different surfaces with which people may come in... 

    Engineering of mature human induced pluripotent stem cell-derived cardiomyocytes using substrates with multiscale topography

    , Article Advanced Functional Materials ; Volume 28, Issue 19 , 2018 ; 1616301X (ISSN) Pour Shahid Saeed Abadi, P ; Garbern, J. C ; Behzadi, S ; Hill, M. J ; Tresback, J. S ; Heydari, T ; Ejtehadi, M. R ; Ahmed, N ; Copley, E ; Aghaverdi, H ; Lee, R. T ; Farokhzad, O. C ; Mahmoudi, M ; Sharif University of Technology
    Wiley-VCH Verlag  2018
    Abstract
    Producing mature and functional cardiomyocytes (CMs) by in vitro differentiation of induced pluripotent stem cells (iPSCs) using only biochemical cues is challenging. To mimic the biophysical and biomechanical complexity of the native in vivo environment during the differentiation and maturation process, polydimethylsiloxane substrates with 3D topography at the micrometer and sub-micrometer levels are developed and used as cell-culture substrates. The results show that while cylindrical patterns on the substrates resembling mature CMs enhance the maturation of iPSC-derived CMs, sub-micrometer-level topographical features derived by imprinting primary human CMs further accelerate both the... 

    Numerical simulation of mixing and heat transfer in an integrated centrifugal microfluidic system for nested-PCR amplification and gene detection

    , Article Sensors and Actuators, B: Chemical ; Volume 283 , 2019 , Pages 831-841 ; 09254005 (ISSN) Naghdloo, A ; Ghazimirsaeed, E ; Shamloo, A ; Sharif University of Technology
    Elsevier B.V  2019
    Abstract
    Nucleic acid amplification via polymerase chain reaction (PCR) is one of the essential and powerful methods used in a myriad of bio-assays in clinical laboratories. Application of microfluidic devices in biologically-related processes like PCR can result in the usage of less volume of reactant samples and reduce the processing time. By implementing PCR systems on centrifugal microfluidic platforms, automation and portability can be easily achieved. Although several methods have been developed, most of them are still dealing with challenges of the required high processing time. This study presents the numerical simulation of a fully automated PCR system with the goal of enhancing the mixing...