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https://www.readbyqxmd.com/read/28103530/an-anti-oxidant-%C3%AE-lipoic-acid-conjugated-oleoyl-sn-phosphatidylcholineas-a-helper-lipid-in-cationic-liposomal-formulations
#1
Priya Dharmalingam, Balakrishna Marrapu, Chandrashekhar Voshavar, Rasagna Nadella, Vignesh Kumar Rangasami, R V Shaji, Salar Abbas, R B N Prasad, Shiva Shanker Kaki, Srujan Marepally
Development of safe non-viral carrier systems for efficient intra-cellular delivery of drugs and genes hold promise in the area of translational research. Liposome based delivery systems have emerged as one of the attractive strategies for efficient delivery of drugs and nucleic acids. To this end, number of investigations was carried on liposomal formulations using lipids for achieving higher efficiency in transfection with lower cytotoxicities. In our efforts to develop safer and efficient liposomal delivery systems, we synthesized a novel anti-oxidant lipid, α-lipoyl, oleyl-sn-phosphatidylcholine (LOPC) and used as a helper lipid in combination with a cationic amphiphile, Di-Stearyl Dihydroxy Ethyl Ammonium Chloride (DSDEAC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) at varying concentrations of LOPC...
January 10, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28102176/synthesis-of-magnetic-cytosine-imprinted-chitosan-nanoparticles
#2
Mei-Hwa Lee, Arti Ahluwalia, Jian-Zhou Chen, Neng-Lang Shih, Hung-Yin Lin
Molecularly imprinted polymer nanoparticles incorporating magnetic nanoparticles (MNPs) have been investigated for their selective adsorption properties. Here we describe the synthesis and characterization of magnetic cytosine-imprinted chitosan nanoparticles (CIPs) for gene delivery. In particular, CIPs carrying the mammalian expression plasmid of enhanced green fluorescent protein were prepared by the co-precipitation of MNPs, chitosan and a template nucleobase (cytosine). The results show that the selective reabsorption of cytosine to magnetic CIPs was at least double that of non-imprinted polymers and other nucleobases (such as adenine and thymine)...
January 19, 2017: Nanotechnology
https://www.readbyqxmd.com/read/28099519/crispr-cas9-mediated-nox4-knockout-inhibits-cell-proliferation-and-invasion-in-hela-cells
#3
Naser Jafari, Hyunju Kim, Rackhyun Park, Liqing Li, Minsu Jang, Andrew J Morris, Junsoo Park, Cai Huang
Increased expression of NOX4 protein is associated with cancer progression and metastasis but the role of NOX4 in cell proliferation and invasion is not fully understood. We generated NOX4 knockout HeLa cell lines using the CRISPR-Cas9 gene editing system to explore the cellular functions of NOX4. After transfection of CRISPR-Cas9 construct, we performed T7 endonuclease 1 assays and DNA sequencing to generate and identify insertion and deletion of the NOX4 locus. We confirmed the knockout of NOX4 by Western blotting...
2017: PloS One
https://www.readbyqxmd.com/read/28095922/long-noncoding-rna-braveheart-promotes-cardiogenic-differentiation-of-mesenchymal-stem-cells-in-vitro
#4
Jingying Hou, Huibao Long, Changqing Zhou, Shaoxin Zheng, Hao Wu, Tianzhu Guo, Quanhua Wu, Tingting Zhong, Tong Wang
BACKGROUND: Mesenchymal stem cells (MSCs) have limited potential of cardiogenic differentiation. In this study, we investigated the influence of long noncoding RNA Braveheart (lncRNA-Bvht) on cardiogenic differentiation of MSCs in vitro. METHODS: MSCs were obtained from C57BL/6 mice and cultured in vitro. Cells were divided into three groups: blank control, null vector control, and lncRNA-Bvht. All three groups experienced exposure to hypoxia (1% O2) and serum deprivation for 24 h, and 24 h of reoxygenation (20% O2)...
January 17, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/28081459/targeted-delivery-of-in-situ-pcr-amplified-sleeping-beauty-transposon-genes-to-cancer-cells-with-lipid-based-nanoparticle-like-protocells
#5
Kun Ma, Duo Fu, Dongli Yu, Changhao Cui, Li Wang, Zhaoming Guo, Chuanbin Mao
A Sleeping Beauty (SB) transposon system is made of a transposon plasmid (containing gene encoding a desired functional or therapeutic protein) and a transposase plasmid (encoding an enzyme capable of cutting and pasting the gene into the host cell genome). It is a kind of natural, nonviral gene delivery vehicle, which can achieve efficient genomic insertion, providing long-term transgenic expression. However, before the SB transposon system could play a role in promoting gene expression, it has to be delivered efficiently first across cell membrane and then into cell nuclei...
January 2, 2017: Biomaterials
https://www.readbyqxmd.com/read/28074350/a-comparative-study-of-the-bone-regenerative-effect-of-chemically-modified-rna-encoding-bmp-2-or-bmp-9
#6
Behnoush Khorsand, Satheesh Elangovan, Liu Hong, Alexander Dewerth, Michael S D Kormann, Aliasger K Salem
Employing cost-effective biomaterials to deliver chemically modified ribonucleic acid (cmRNA) in a controlled manner addresses the high cost, safety concerns, and lower transfection efficiency that exist with protein and gene therapeutic approaches. By eliminating the need for nuclear entry, cmRNA therapeutics can potentially overcome the lower transfection efficiencies associated with non-viral gene delivery systems. Here, we investigated the osteogenic potential of cmRNA-encoding BMP-9, in comparison to cmRNA-encoding BMP-2...
January 10, 2017: AAPS Journal
https://www.readbyqxmd.com/read/28051330/g-protein-alpha-s-subunit-promotes-cell-proliferation-of-renal-cell-carcinoma-with-involvement-of-protein-kinase-a-signaling
#7
Bo Zhang, Nan Sun, Xin Mu, Lei Zhi, Lei Zhai, Yuan Jiang, Zheng Fu, Zhi Yao
Heterotrimeric G proteins, which are composed of Gα and Gβγ subunits, transduce signals sensed by the coupled surface receptors. Aberrant expressions of G proteins have been observed in many cancer types. This study aimed to determine the expression level of the stimulatory G protein alpha S subunit (Gαs, the main transcript encoded by the GNAS locus) and its biological function in renal cell carcinoma (RCC). Western blotting and quantitative reverse transcription-PCR results show that Gαs expression dramatically increased in RCC cell lines (ACHN, GRC-1, and 786-O) compared to normal renal epithelial cells HK-2...
January 4, 2017: DNA and Cell Biology
https://www.readbyqxmd.com/read/28043862/rapid-and-facile-quantitation-of-polyplex-endocytic-trafficking
#8
Mihael Lazebnik, Daniel W Pack
Design of safe and effective synthetic nucleic acid delivery vectors such as polycation/DNA or polycation/siRNA complexes (polyplexes) will be facilitated by quantitative understanding of the mechanisms by which such materials escort cargo from the cell surface to the nucleus. In particular, the mechanisms of cellular internalization by various endocytosis pathways and subsequent endocytic vesicle trafficking have been shown to strongly affect nucleic acid delivery efficiency. Fluorescence microscopy and subcellular fractionation methods are commonly employed to follow intracellular trafficking of biomolecules and nanoparticulate delivery systems such as polyplexes...
December 31, 2016: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28042316/localized-delivery-of-shrna-against-phd2-protects-the-heart-from-acute-myocardial-infarction-through-ultrasound-targeted-cationic-microbubble-destruction
#9
Li Zhang, Zhenxing Sun, Pingping Ren, Manjie You, Jing Zhang, Lingyun Fang, Jing Wang, Yihan Chen, Fei Yan, Hairong Zheng, Mingxing Xie
Hypoxia-inducible factor 1α (HIF-1α) plays a critical protective role in ischemic heart disease. Under normoxic conditions, HIF-1α was degraded by oxygen-dependent prolyl hydroxylase-2 (PHD2). Gene therapy has become a promising strategy to inhibit the degradation of HIF-1α and to improve cardiac function after ischemic injury. However, conventional gene delivery systems are difficult to achieve a targeted and localized gene delivery into the ischemic myocardia. Here, we report the localized myocardial delivery of shRNA against PHD2 through ultrasound-targeted microbubble destruction (UTMD) for protection the heart from acute myocardial infarction...
2017: Theranostics
https://www.readbyqxmd.com/read/28038351/microrna-29-impairs-the-early-phase-of-reprogramming-process-by-targeting-active-dna-demethylation-enzymes-and-wnt-signaling
#10
Mariane Serra Fráguas, Reto Eggenschwiler, Jeannine Hoepfner, Josiane Lilian Dos Santos Schiavinato, Rodrigo Haddad, Lucila Habib Bourguignon Oliveira, Amélia Góes Araújo, Marco Antônio Zago, Rodrigo Alexandre Panepucci, Tobias Cantz
Somatic cell reprogramming by transcription factors and other modifiers such as microRNAs has opened broad avenues for the study of developmental processes, cell fate determination, and interplay of molecular mechanisms in signaling pathways. However, many of the mechanisms that drive nuclear reprogramming itself remain yet to be elucidated. Here, we analyzed the role of miR-29 during reprogramming in more detail. Therefore, we evaluated miR-29 expression during reprogramming of fibroblasts transduced with lentiviral OKS and OKSM vectors and we show that addition of c-MYC to the reprogramming factor cocktail decreases miR-29 expression levels...
December 19, 2016: Stem Cell Research
https://www.readbyqxmd.com/read/28036161/use-of-primary-and-secondary-polyvinylamines-for-efficient-gene-transfection
#11
Mathilde Dréan, Antoine Debuigne, Cristine Gonçalves, Christine Jérôme, Patrick Midoux, Jutta Rieger, Philippe Guegan
Gene transfection with polymeric carrier remains a challenge, particularly high transfection levels combined with low toxicity are hard to achieve. We herein revisit polyvinylamines, an old and neglected family of cationic polymers. They can be readily obtained by controlled hydrolysis of polyvinylamides prepared through (controlled) radical polymerization. A series of tailor-made and well-defined polyvinylamines bearing primary amino groups, and poly(N-methylvinylamine) bearing secondary amines, were evaluated for the transfection of cells with pDNA as a function of their molar mass, molar mass distribution and degree of deacetylation...
December 30, 2016: Biomacromolecules
https://www.readbyqxmd.com/read/28033108/tissue-factor-promotes-breast-cancer-stem-cell-activity-in-vitro
#12
Hudhaifah Shaker, Hannah Harrison, Robert Clarke, Goran Landberg, Nigel J Bundred, Henri H Versteeg, Cliona C Kirwan
Cancer stem cells (CSCs) are a subpopulation of cells that can self-renew and initiate tumours. The clotting-initiating protein Tissue Factor (TF) promotes metastasis and may be overexpressed in cancer cells with increased CSC activity. We sought to determine whether TF promotes breast CSC activity in vitro using human breast cancer cell lines. TF expression was compared in anoikis-resistant (CSC-enriched) and unselected cells. In cells sorted into of TF-expressing and TF-negative (FACS), and in cells transfected to knockdown TF (siRNA) and overexpress TF (cDNA), CSC activity was compared by (i) mammosphere forming efficiency (MFE) (ii) holoclone colony formation (Hc) and (iii) ALDH1 activity...
December 13, 2016: Oncotarget
https://www.readbyqxmd.com/read/28027876/ercc2-xpd-lys751gln-alter-dna-repair-efficiency-of-platinum-induced-dna-damage-through-p53-pathway
#13
Guopei Zhang, Yangyang Guan, Yuejiao Zhao, Tahar van der Straaten, Sha Xiao, Ping Xue, Guolian Zhu, Qiufang Liu, Yuan Cai, Cuihong Jin, Jinghua Yang, Shengwen Wu, Xiaobo Lu
Platinum-based treatment causes Pt-DNA adducts which lead to cell death. The platinum-induced DNA damage is recognized and repaired by the nucleotide excision repair (NER) system of which ERCC2/XPD is a critical enzyme. Single nucleotide polymorphisms in ERCC2/XPD have been found to be associated with platinum resistance. The aim of the present study was to investigate whether ERCC2/XPD Lys751Gln (rs13181) polymorphism is causally related to DNA repair capacity of platinum-induced DNA damage. First, cDNA clones expressing different genotypes of the polymorphism was transfected to an ERCC2/XPD defective CHO cell line (UV5)...
December 24, 2016: Chemico-biological Interactions
https://www.readbyqxmd.com/read/28025049/polylysine-modified-polyethylenimine-pei-pll-mediated-vegf-gene-delivery-protects-dopaminergic-neurons-in-cell-culture-and-in-rat-models-of-parkinson-s-disease-pd
#14
Muhammad Abid Sheikh, Yousra Saeed Malik, Zhenkai Xing, Zhaopei Guo, Huayu Tian, Xiaojuan Zhu, Xuesi Chen
Parkinson's Disease (PD) is a chronic neurodegenerative disorder characterized by motor deficits which result from the progressive loss of dopaminergic neurons. Gene therapy using growth factors such as VEGF seems to be a viable approach for potential therapeutic treatment of PD. In this study, we utilized a novel non-viral gene carrier designated as PEI-PLL synthesized by our laboratory to deliver VEGF gene to study its effect by using both cell culture as well as animal models of PD. For cell culture experiments, we utilized 6-hydroxydopamine (6-OHDA) mediated cell death model of MN9D cells following transfection with either a control plasmid or VEGF expressing plasmid...
December 23, 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/28024496/-construction-of-lentiviral-vector-over-expressing-meg3-and-its-effect-on-xg-7-cell-apoptosis
#15
Yi-Kun Zhang, Hua Wang, Feng-Jun Xiao, Xiao-Yan Zhang, Pei-Lin Liu, Quan-Xing Shi, Zhao Yin, Yan Lei, Li-Sheng Wang
OBJECTIVE: To construct a recombinant lentiviral expression vectors carrying MEG3 and to evaluate its effects on XG-7 cell apoptosis. METHODS: A full-length genomic fragment of human MEG3 was cloned from the pcDNA3.0-MEG3 packaging plasmid and was amplified by PCR. New restriction sites were introduced to be blunted with T4 DNA Ligase. The sequence of the amplified segments was sub-cloned into lentivirus expression vector pCDH-EF1-MCS-T2A-copGFP.The recombined lentiviral expression vector was transfected into 293T cells...
December 2016: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/28013079/development-of-dual-fluorescence-cell-based-biosensors-for-detecting-the-influence-of-environmental-factors-on-nanoparticle-toxicity
#16
Junting Liang, Xuanyu Wang, Luzhi Li, Shengmin Xu, Jiang Jiang, Lijun Wu, Guoping Zhao, Shaopeng Chen
With the expanding use of engineered nanoparticles (NPs), development of a high-throughput, sensitive method for evaluating NP safety is important. In this study, we developed cell-based biosensors to efficiently and conveniently monitor NP toxicity. The biosensor cells were obtained by transiently transfecting human cells with biosensor plasmids containing a mCherry gene regulated by an inducible promoter [an activator protein 1 (AP-1) promoter, an interleukin 8 (IL8) promoter, or a B cell translocation gene 2 (BTG2) promoter], with an enhanced green-fluorescent protein gene driven by the cytomegalovirus promoter as the internal control...
December 18, 2016: Chemosphere
https://www.readbyqxmd.com/read/28011343/design-and-characterization-of-a-magnetite-pei-multifunctional-nanohybrid-as-non-viral-vector-and-cell-isolation-system
#17
Raúl Megías, Maialen Arco, Jesús Ciriza, Laura Saenz Del Burgo, Gustavo Puras, Margarita López-Viota, Ángel V Delgado, Jon P Dobson, José L Arias, José L Pedraz
It is described the reproducible formulation and complete physicochemical characterization of nanohybrids based on magnetite (Fe3O4) cores embedded within a polyethylenimine (PEI) matrix. Particle size, surface electrical charge, X-ray diffraction and Fourier transform infrared spectroscopy (FTIR) analyses, and magnetic field-responsive behaviour characterizations defined that the 4:3 (Fe3O4:PEI) weight proportion led to the best production performances of magnetically responsive nanocomposites in which the magnetic nuclei are completely covered by the polymeric shell...
December 21, 2016: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/27998765/mir-223-modulates-hepatocellular-carcinoma-cell-proliferation-through-promoting-apoptosis-via-the-rab1-mediated-mtor-activation
#18
Zheng Dong, Ruizhao Qi, Xiaodong Guo, Xin Zhao, Yinyin Li, Zhen Zeng, Wenlin Bai, Xiujuan Chang, Liyan Hao, Yan Chen, Min Lou, Zhiwei Li, Yinying Lu
Hepatocellular carcinoma (HCC) is a common digestive malignancy. MiR-223, a well-identified miRNA, exhibits diverse properties in different cancers. In this study, we demonstrated that miR-223 could suppress cell growth and promote apoptosis in HepG2 and Bel-7402 HCC cell lines. We screened and identified a novel miR-223 target, Ras-related protein Rab-1(Rab1). Upregulation of miR-223 would specifically and markedly down-regulate Rab1 expression. In addition, miR-223-overexpressing subclones showed significant cell growth inhibition by increasing cell apoptosis in HepG2 and Bel-7402 cells...
December 18, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27996099/preliminary-study-of-a-novel-transfection-modality-for-in-vivo-sirna-delivery-to-vocal-fold-fibroblasts
#19
Iv Kraja, Renjie Bing, Nao Hiwatashi, Bernard Rousseau, Danielle Nalband, Kent Kirshenbaum, Ryan C Branski
OBJECTIVE: An obstacle to clinical use of RNA-based gene suppression is instability and inefficiency of current delivery modalities. Nanoparticle delivery likely holds great promise, but the kinetics and transfection conditions must be optimized prior to in vivo utility. We investigated a RNA nanoparticle complex incorporating a lipitoid transfection reagent in comparison to a commercially available reagent. STUDY DESIGN: In vitro. METHODS: We investigated which variables influence transfection efficiency of lipitoid oligomers and a commercially available reagent across species, in vitro...
December 20, 2016: Laryngoscope
https://www.readbyqxmd.com/read/27992192/heparin-enhances-transfection-in-concert-with-a-trehalose-based-polycation-with-challenging-cell-types
#20
William S Boyle, Kyle Senger, Jakub Tolar, Theresa M Reineke
Improving the delivery of nucleic acids to diverse tissue types in culture is important for translating genome editing for regenerative cell therapies. Herein, we examine the effect of transfection media additives, such as the sulfated glycosaminoglycan heparin, in dramatically increasing pDNA delivery efficiency and transgene expression in a wide variety of cell types. Polyplexes formed by combining pDNA and Tr4, a cationic glycopolymer containing repeated trehalose and pentaethylenetetramine groups, were treated with low concentrations of heparin prior to in vitro transfection with plasmid DNA...
December 19, 2016: Biomacromolecules
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