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https://www.readbyqxmd.com/read/28717157/cellular-uptake-pathways-of-sepiolite-nanofibers-and-dna-transfection-improvement
#1
Fidel Antonio Castro-Smirnov, Jeanne Ayache, Jean-Rémi Bertrand, Elodie Dardillac, Eric Le Cam, Olivier Piétrement, Pilar Aranda, Eduardo Ruiz-Hitzky, Bernard S Lopez
Sepiolite is a nanofibrous natural silicate that can be used as a nanocarrier because it can be naturally internalized into mammalian cells, due to its nano-size dimension. Therefore, deciphering the mechanisms of sepiolite cell internalization constitutes a question interesting biotechnology, for the use of sepiolite as nanocarrier, as well as environmental and public health concerns. Though it is low, the perfectly stable and natural intrinsic fluorescence of sepiolite nanofibers allows to follow their fate into cells by specifically sensitive technics...
July 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28714224/codelivery-for-paclitaxel-and-bcl-2-conversion-gene-by-phb-pdmaema-amphiphilic-cationic-copolymer-for-effective-drug-resistant-cancer-therapy
#2
Xiaoyuan Wang, Sing Shy Liow, Qiaoqiong Wu, Chuang Li, Cally Owh, Zibiao Li, Xian Jun Loh, Yun-Long Wu
Antiapoptotic Bcl-2 protein's upregulated expression is a key reason for drug resistance leading to failure of chemotherapy. In this report, a series of biocompatible amphiphilic cationic poly[(R)-3-hydroxybutyrate] (PHB)-b-poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) copolymer, comprising hydrophobic PHB block and cationic PDMAEMA block, is designed to codeliver hydrophobic chemotherapeutic paclitaxel and Bcl-2 converting gene Nur77/ΔDBD with enhanced stability, due to the micelle formation by hydrophobic PHB segment...
July 17, 2017: Macromolecular Bioscience
https://www.readbyqxmd.com/read/28710005/cationic-microbubbles-and-antibiotic-free-miniplasmid-for-sustained-ultrasound-mediated-transgene-expression-in-liver
#3
Simona Manta, Gilles Renault, Anthony Delalande, Olivier Couture, Isabelle Lagoutte, Johanne Seguin, Franck Lager, Pascal Houzé, Patrick Midoux, Michel Bessodes, Daniel Scherman, Michel-Francis Bureau, Corinne Marie, Chantal Pichon, Nathalie Mignet
Despite the increasing number of clinical trials in gene therapy, no ideal methods still allow non-viral gene transfer in deep tissues such as the liver. We were interested in ultrasound (US)-mediated gene delivery to provide long term liver expression. For this purpose, new positively charged microbubbles were designed and complexed with pFAR4, a highly efficient small length miniplasmid DNA devoid of antibiotic resistance sequence. Sonoporation parameters, such as insonation time, acoustic pressure and duration of plasmid injection were controlled under ultrasound imaging guidance...
July 11, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28708803/hydrodynamic-il10-gene-transfer-in-human-colon-results-from-an-ex-vivo-study-with-potential-clinical-application-in-crohn-s-disease
#4
Matteo Frasson, Luis Sendra, Antonio Miguel, Maria José Herrero, Eva Montalvá, Rafael López-Andújar, Juan Martínez-Pastor, Luis Martí-Bonmatí, Eduardo García Granero, Salvador Aliño
BACKGROUND: The aim of this work is to evaluate the efficacy of hydrodynamic venous IL10 gene delivery to "ex vivo" human colon segments and to determine its potential interest in Crohn's disease treatment. METHODS: Twenty human colon segments were obtained from surgical resections. Hydrodynamic transfection through the main vein of the pedicle with 50 mL of hIL10 plasmid (20 μg/mL) solution was performed on 13 of them. Tissue sections were cultured and DNA, RNA, and protein copies were determined after 1, 2, and 4 days...
August 2017: Inflammatory Bowel Diseases
https://www.readbyqxmd.com/read/28707662/-c-terminal-lysosome-targeting-domain-of-cd63-modifies-cellular-localization-of-rabies-virus-glycoprotein
#5
E S Starodubova, Y V Kuzmenko, A A Latanova, O V Preobrazhenskaya, V L Karpov
The glycoprotein of rabies virus is the central antigen elicited the immune response to infection; therefore, the majority of developing anti-rabies vaccines are based on this protein. In order to increase the efficacy of DNA immunogen encoding rabies virus glycoprotein, the construction of chimeric protein with the CD63 domain has been proposed. The CD63 is a transmembrane protein localized on the cell surface and in lysosomes. The lysosome targeting motif GYEVM is located at its C-terminus. We used the domain that bears this motif (c-CD63) to generate chimeric glycoprotein in order to relocalize it into lysosomes...
May 2017: Molekuliarnaia Biologiia
https://www.readbyqxmd.com/read/28706136/swine-irf3-irf7-attenuates-inflammatory-responses-through-tlr4-signaling-pathway
#6
Pei-Ge Chen, Yan-Jing Guan, Guang-Ming Zha, Xian-Qin Jiao, He-Shui Zhu, Cheng-Yu Zhang, Yue-Ying Wang, He-Ping Li
To explore the role of IRF3/IRF7 during inflammatory responses, we investigated the effects of swine IRF3/IRF7 on TLR4 signaling pathway and inflammatory factors expression in porcine kidney epithelial PK15 cell lines. We successfully constructed eukaryotic vectors PB-IRF3 and PB-IRF7, transfected these vectors into PK15 cells and observed GFP under a fluorescence microscope. In addition, RT-PCR was also used to detect transfection efficiency. We found that IRF3/IRF7 was efficiently overexpressed in PK15 cells...
June 28, 2017: Oncotarget
https://www.readbyqxmd.com/read/28701147/combinational-use-of-lipid-based-reagents-for-efficient-transfection-of-primary-fibroblasts-and-hepatoblasts
#7
Kazuhiro Ishiguro, Osamu Watanabe, Masanao Nakamura, Takeshi Yamamura, Masanobu Matsushita, Hidemi Goto, Yoshiki Hirooka
Commercially available lipid-based transfection reagents are widely used to deliver DNA to cells. However, these lipid-based transfection reagents show poor gene transfer efficiency in primary cells. Here, we demonstrate a simple method to improve gene transfer efficiency in primary fibroblasts and hepatoblasts using a combination of lipid-based transfection reagents. Our data show that combined use of Lipofectamine LTX and FuGENE HD increases the efficiency of gene transfer compared with the use of either reagent alone, and this combination achieves the best result of any pairwise combination of Lipofectamine LTX, FuGENE HD, TransFectin, and Fibroblast Transfection Reagent...
July 1, 2017: BioTechniques
https://www.readbyqxmd.com/read/28698591/molecular-dynamics-study-on-the-mechanism-of-polynucleotide-encapsulation-by-chitosan
#8
Jia-Wei Shen, Jiachen Li, Zhennan Zhao, Li Zhang, Guoteng Peng, Lijun Liang
The safe and effective delivery of therapeutic genes into target cell interiors is of great importance in gene therapy. Chitosan has been extensively studied as a gene delivery carrier due to its good biocompatibility and biodegradability. Understanding the atomic interaction mechanism between chitosan and DNA is important in the design and application of chitosan-based drug and gene delivery systems. In this work, the interactions between single-stranded polynucleotides and different types of chitosan were systematically investigated by using molecular dynamics (MD) simulation...
July 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28697439/targeting-dendritic-cells-for-the-treatment-of-autoimmune-disorders
#9
REVIEW
C Sousa, I Pereira, A C Santos, C Carbone, A B Kovačević, A M Silva, E B Souto
In the last decades, it has been recognized that extracellular vesicles (EVs) are not only cell debris with no biological role, but instead they play a key role in information exchange between cells either in health and disease conditions. EVs exhibit indeed their biological role in a pleiotropic manner. They can modulate immune responses through the activation, transfer or removal of surface receptors on target cells, the removal of cytolytic components such as membrane attack complexes, and the transfer of signaling molecules/effectors, such as nucleic acid species, infectious particles, and oncogenes...
June 30, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28689990/graphene-oxide-nanosheets-in-complex-with-cell-penetrating-peptides-for-oligonucleotides-delivery
#10
Moataz Dowaidar, Hani Nasser Abdelhamid, Mattias Hällbrink, Xiaodong Zou, Ülo Langel
A new strategy for gene transfection using the nanocarrier of cell penetrating peptides (CPPs; PepFect14 (PF14) or PepFect14 (PF14) (PF221)) in complex with graphene oxide (GO) is reported. GO complexed with CPPs and plasmid (pGL3), splice correction oligonucleotides (SCO) or small interfering RNA (siRNA) are performed. Data show adsorption of CPPs and oligonucleotides on the top of the graphenic lamellar without any observed change of the particle size of GO. GO mitigates the cytotoxicity of CPPs and improves the material biocompatibility...
July 6, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28686452/immunostimulatory-properties-of-lipid-modified-cpg-oligonucleotides
#11
Chunsong Yu, Myunggi An, Meng Li, Haipeng Liu
Innate immune responses recognizing pathogen associated molecular patterns play important roles in adaptive immunity. As such, ligands which mimic the conserved products of microbial and activate innate immunity are widely used as adjuvants for vaccines. Synthetic single strand oligodeoxynucleotides (ODNs) containing unmethylated cytosine-guanine (CpG) motifs which bind Toll-like receptor 9 (TLR9) are powerful molecular adjuvants, potentiating both humoral and cellular responses. However, CpG ODN's in vitro potency has not been translated to in vivo settings primarily due to issues associated with delivery and toxicity...
July 7, 2017: Molecular Pharmaceutics
https://www.readbyqxmd.com/read/28685239/a-biodesign-approach-to-obtain-high-yields-of-biosimilars-by-anti-apoptotic-cell-engineering-a-case-study-to-increase-the-production-yield-of-anti-tnf-alpha-producing-recombinant-cho-cells
#12
Sultan Gulce Iz, Muge Anil Inevi, Pelin Saglam Metiner, Duygu Ayyildiz Tamis, Nazli Kisbet
Recent developments in medical biotechnology have facilitated to enhance the production of monoclonal antibodies (mAbs) and recombinant proteins in mammalian cells. Human mAbs for clinical applications have focused on three areas, particularly cancer, immunological disorders, and infectious diseases. Tumor necrosis factor alpha (TNF-α), which has both proinflammatory and immunoregulatory functions, is an important target in biopharmaceutical industry. In this study, a humanized anti-TNF-α mAb producing stable CHO cell line which produces a biosimilar of Humira (adalimumab) was used...
July 6, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28682874/in-vitro-effect-of-microrna-107-targeting-dkk-1-by-regulation-of-wnt-%C3%AE-catenin-signaling-pathway-in-osteosarcoma
#13
Zhi-Cai Zhang, Jian-Xiang Liu, Zeng-Wu Shao, Fei-Fei Pu, Bai-Chuan Wang, Qiang Wu, Yu-Kun Zhang, Xian-Lin Zeng, Xiao-Dong Guo, Shu-Hua Yang, Tong-Chuan He
BACKGROUND: The aim of the study was to explore the effects of microRNA-107 (miR-107) by targeting Dkk-1 on osteosarcoma (OS) via the Wnt/β-catenin signaling pathway. METHODS: OS and adjacent tissues were collected from 67 patients diagnosed with OS. Expressions of miR-107, Dkk-1, LRP5, β-catenin, and c-Myc were detected by the quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. The dual-luciferase reporter gene assay was performed to observe the relationship between miR-107 and Dkk-1...
July 2017: Medicine (Baltimore)
https://www.readbyqxmd.com/read/28679425/overexpression-of-c-met-in-bone-marrow-mesenchymal-stem-cells-improves-their-effectiveness-in-homing-and-repair-of-acute-liver-failure
#14
Kun Wang, Yuwen Li, Tiantian Zhu, Yongting Zhang, Wenting Li, Wenyu Lin, Jun Li, Chuanlong Zhu
BACKGROUND: Transplantation of bone marrow-derived mesenchymal stem cells (BMSCs) has emerged as a novel therapy for acute liver failure (ALF). However, the homing efficiency of BMSCs to the injured liver sites appears to be poor. In this study, we aimed to determine if overexpression of c-Met in BMSCs could promote the homing ability of BMSCs to rat livers affected by ALF. METHODS: Overexpression of c-Met in BMSCs (c-Met-BMSCs) was attained by transfection of naive BMSCs with the lenti-c-Met-GFP...
July 5, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/28679128/comparison-of-intracoronary-and-intravenous-ultrasound-targeted-microbubble-destruction-mediated-ang1-gene-transfection-on-left-ventricular-remodeling-in-canines-with-acute-myocardial-infarction
#15
Sheng Cao, Qing Zhou, Jin-Ling Chen, Jing-Jing Cui, Ying-Guang Shan, Bo Hu, Rui-Qiang Guo
Intravenous ultrasound-targeted microbubble destruction (IV-UTMD) has made distinct but limited progress in gene therapy. Intracoronary (IC) injection may lead to more gene transfection than IV injection. This study compared the therapeutic effects of IC-UTMD-mediated and conventional IV-UTMD-mediated gene transfection in acute myocardial infarction (MI). A canine MI model was successfully established through transcatheter coronary artery embolism, and the animals were divided into several treatment groups: IC injection with UTMD and the negative control plasmid (IC-UTMD); IC injection of the angiopoietin 1 (Ang1) plasmid (IC-Ang1); IC injection with UTMD and the Ang1 plasmid (IC-UTMD-Ang1); and IV injection with UTMD and the Ang1 plasmid (IV-UTMD-Ang1)...
July 2017: Journal of Cardiovascular Pharmacology
https://www.readbyqxmd.com/read/28674801/high-efficiency-transfection-of-glioblastoma-cells-and-a-simple-spheroid-migration-assay
#16
Carsten Hagemann, Diana Amend, Almuth F Kessler, Thomas Linsenmann, Ralf-Ingo Ernestus, Mario Löhr
Despite international research efforts, patients with glioblastoma multiforme (GBM)-the most common malignant brain tumors in adults-exhibit a very unfavorable prognosis. Their aggressive local growth pattern and increased invasiveness, due to a high motility of the tumor cells, hamper treatment. However, the molecular mechanisms regulating glioblastoma cell migration are still elusive. Here, we describe the combination of a highly efficient cell transfection by Nucleofection(®) technology and the generation of spheroids from these transfected glioblastoma cell lines...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28672998/role-of-angiopoietin-2-in-the-cardioprotective-effect-of-fibroblast-growth-factor-21-on-ischemia-reperfusion-induced-injury-in-h9c2-cardiomyocytes
#17
Shuoqiang Hu, Shujun Cao, Jinghua Liu
Fibroblast growth factor 21 (FGF21) exerts a protective effect in ischemia/reperfusion (I/R)-induced cardiac injury. However, the exact molecular mechanism underlying the FGF21 action remains unclear. The present study aimed to evaluate the role of angiopoietin-2 (Angpt2) in the cardioprotective effect of FGF21. For this purpose, the H9C2 cell line was subjected to simulated I/R or aerobic conditions with or without FGF21 administration. Certain groups were also transfected with Angpt2 small interfering RNA (siRNA)...
July 2017: Experimental and Therapeutic Medicine
https://www.readbyqxmd.com/read/28670930/topical-delivery-of-trpsirna-loaded-solid-lipid-nanoparticles-confer-reduced-pain-sensation-via-trpv1-silencing-in-rats
#18
Gaurav Sharma, Kanwaljit Chopra, Sanjeev Puri, Mahendra Bishnoi, Parveen Rishi, Indu P Kaur
Present work describes a novel composition for encapsulating TRPsiRNA (TRPV1 targeting siRNA) within lipid-matrix (4:1::glyceryl behnate:stearic acid) of SLNs, using suitably modified cold high-pressure homogenization technique. Optimization of the method and composition conducted using calf-thymus DNA (ctDNA), to avoid cost of TRPsiRNA molecules, resulted in small size (d50=50-100nm) and high entrapment (77.22-98.5%). Complete masking of extreme negative charge of both ctDNA (-34.50mV) and TRPsiRNA (-23.98mV) upon encapsulation in SLNs without employing cationic components is reported herein for the first time...
July 3, 2017: Journal of Drug Targeting
https://www.readbyqxmd.com/read/28665509/direct-reprogramming-of-human-suspension-cells-into-mesodermal-cell-lineages-via-combined-magnetic-targeting-and-photothermal-stimulation-by-magnetic-graphene-oxide-complexes
#19
Min-Yu Chiang, Yi-Zhen Lin, Shwu-Jen Chang, Woei-Cherng Shyu, Huai-En Lu, San-Yuan Chen
Suspension cells can provide a source of cells for cellular reprogramming, but they are difficult to transfect by nonviral vectors. An efficient and safe nonviral vector (GO-Fe3 O4 -PEI complexes) based on iron oxide nanoparticle (Fe3 O4 )-decorated graphene oxide (GO) complexed with polyethylenimine (PEI) for the first time is developed for delivering three individual episomal plasmids (pCXLE-hOCT3/4-shp53, pCXLE-hSK, and pCXLE-hUL) encoding pluripotent-related factors of Oct3/4, shRNA against p53, Sox2, Klf4, L-Myc, and Lin28 into human peripheral blood mononuclear cells (PBMCs) simultaneously...
June 30, 2017: Small
https://www.readbyqxmd.com/read/28656285/lkb1-promotes-radioresistance-in-esophageal-cancer-cells-exposed-to-radiation-by-suppression-of-apoptosis-and-activation-of-autophagy-via-the-ampk-pathway
#20
Qing He, Jing Li, Feng Dong, Chuanshu Cai, Xi Zou
Liver kinase B (LKB) 1 acts as a tumor suppressor in a broad spectrum of human cancers, and is important in chemoradiotherapy treatment of various tumor types. However, the potential function of LKB1 in esophageal cancer radiotherapy remains to be elucidated. The aim of the present study was to investigate the role of LKB1 in radiosensitivity of esophageal cancer in vivo and in vitro, and to explore its molecular mechanism. Eca‑109 cells transfected with LKB1 overexpression plasmid were xenografted into nude mice and subjected to irradiation and it was observed that the tumor volume was significantly increased in LKB1‑overexpressed tumors compared with that of the control tumors...
August 2017: Molecular Medicine Reports
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