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https://www.readbyqxmd.com/read/27920148/epilepsy-associated-kcnq2-channels-regulate-multiple-intrinsic-properties-of-layer-2-3-pyramidal-neurons
#1
Zachary Niday, Virginia E Hawkins, Heun Soh, Daniel K Mulkey, Anastasios V Tzingounis
: KCNQ2 potassium channels are critical for normal brain function, as both loss- and gain-of-function KCNQ2 variants can lead to various forms of neonatal epilepsy. Despite recent progress, the full spectrum of consequences as a result of KCNQ2 dysfunction in neocortical pyramidal neurons is still unknown. Here, we report that conditional ablation of Kcnq2 from mouse neocortex leads to hyperexcitability of layer 2/3 (L2/3) pyramidal neurons, exhibiting an increased input resistance and action potential frequency, as well as a reduced medium afterhyperpolarization (mAHP), a conductance partly mediated by KCNQ2 channels...
December 5, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/27918590/effects-of-circular-dna-length-on-transfection-efficiency-by-electroporation-into-hela-cells
#2
Benjamin D Hornstein, Dany Roman, Lirio M Arévalo-Soliz, Melinda A Engevik, Lynn Zechiedrich
The ability to produce extremely small and circular supercoiled vectors has opened new territory for improving non-viral gene therapy vectors. In this work, we compared transfection of supercoiled DNA vectors ranging from 383 to 4,548 bp, each encoding shRNA against GFP under control of the H1 promoter. We assessed knockdown of GFP by electroporation into HeLa cells. All of our vectors entered cells in comparable numbers when electroporated with equal moles of DNA. Despite similar cell entry, we found length-dependent differences in how efficiently the vectors knocked down GFP...
2016: PloS One
https://www.readbyqxmd.com/read/27914928/mathematical-model-for-transport-of-dna-plasmids-from-the-external-medium-up-to-the-nucleus-by-electroporation
#3
M Leguèbe, M G Notarangelo, M Twarogowska, R Natalini, C Poignard
We propose a mathematical model for the transport of DNA plasmids from the extracellular matrix up to the cell nucleus. The model couples two phenomena: the electroporation process, describing the cell membrane permeabilization to plasmids and the intracellular transport enhanced by the presence of microtubules. Numerical simulations of cells with arbitrary geometry, in 2D and 3D, and a network of microtubules show numerically the importance of the microtubules and the electroporation on the effectiveness of the DNA transfection, as observed by previous biological data...
November 30, 2016: Mathematical Biosciences
https://www.readbyqxmd.com/read/27913835/preventive-dna-vaccination-against-cea-expressing-tumors-with-anti-idiotypic-scfv6-c4-dna-in-cea-expressing-transgenic-mice
#4
Priscila M A Denapoli, Bianca F Zanetti, Adara A Dos Santos, Jane Z de Moraes, Sang W Han
Carcinoembryonic antigen (CEA) is expressed during embryonic life and in low level during adult life. Consequently, the CEA is recognized by the immune system as a self-antigen and thus CEA-expressing tumors are tolerated. Previously, we constructed a single chain variable fragment using the 6.C4 (scFv6.C4) hybridoma cell line, which gave rise to antibodies able to recognize CEA when C57/Bl6 mice were immunized. Here, the scFv6.C4 ability to prevent the CEA-expressing tumor growth was assessed in CEA-expressing transgenic mice CEA2682...
December 2, 2016: Cancer Immunology, Immunotherapy: CII
https://www.readbyqxmd.com/read/27912985/immunogenicity-and-malaria-transmission-reducing-potency-of-pfs48-45-and-pfs25-encoded-by-dna-vaccines-administered-by-intramuscular-electroporation
#5
Dibyadyuti Datta, Geetha P Bansal, Dietlind L Gerloff, Barry Ellefsen, Drew Hannaman, Nirbhay Kumar
Pfs48/45 and Pfs25 are leading candidates for the development of Plasmodium falciparum transmission blocking vaccines (TBV). Expression of Pfs48/45 in the erythrocytic sexual stages and presentation to the immune system during infection in the human host also makes it ideal for natural boosting. However, it has been challenging to produce a fully folded, functionally active Pfs48/45, using various protein expression platforms. In this study, we demonstrate that full-length Pfs48/45 encoded by DNA plasmids is able to induce significant transmission reducing immune responses...
November 29, 2016: Vaccine
https://www.readbyqxmd.com/read/27911903/transfected-babesia-bovis-expressing-a-tick-gst-as-a-live-vector-vaccine
#6
Daiane P Oldiges, Jacob M Laughery, Nelson Junior Tagliari, Ronaldo Viana Leite Filho, William C Davis, Itabajara da Silva Vaz, Carlos Termignoni, Donald P Knowles, Carlos E Suarez
The Rhipicephalus microplus tick is a notorious blood-feeding ectoparasite of livestock, especially cattle, responsible for massive losses in animal production. It is the main vector for transmission of pathogenic bacteria and parasites, including Babesia bovis, an intraerythrocytic apicomplexan protozoan parasite responsible for bovine Babesiosis. This study describes the development and testing of a live B. bovis vaccine expressing the protective tick antigen glutathione-S-transferase from Haemaphysalis longicornis (HlGST)...
December 2016: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/27909701/immunological-quality-and-performance-of-tumor-vessel-targeting-car-t-cells-prepared-by-mrna-ep-for-clinical-research
#7
Kanako Inoo, Ryo Inagaki, Kento Fujiwara, Shigemi Sasawatari, Takashi Kamigaki, Shinsaku Nakagawa, Naoki Okada
We previously reported that tumor vessel-redirected T cells, which were genetically engineered with chimeric antigen receptor (CAR) specific for vascular endothelial growth factor receptor 2 (VEGFR2), demonstrated significant antitumor effects in various murine solid tumor models. In the present study, we prepared anti-VEGFR2 CAR-T cells by CAR-coding mRNA electroporation (mRNA-EP) and analyzed their immunological characteristics and functions for use in clinical research. The expression of anti-VEGFR2 CAR on murine and human T cells was detected with approximately 100% efficiency for a few days, after peaking 6-12 hours after mRNA-EP...
2016: Molecular Therapy Oncolytics
https://www.readbyqxmd.com/read/27907195/novel-aryl-hydrocarbon-receptor-agonist-suppresses-migration-and-invasion-of-breast-cancer-cells
#8
Hamza Hanieh, Omar Mohafez, Villianur Ibrahim Hairul-Islam, Abdullah Alzahrani, Mohammad Bani Ismail, Krishnaraj Thirugnanasambantham
BACKGROUND: Despite the remarkable progress to fight against breast cancer, metastasis remains the dominant cause of treatment failure and recurrence. Therefore, control of invasiveness potential of breast cancer cells is crucial. Accumulating evidences suggest Aryl hydrocarbon receptor (Ahr), a helix-loop-helix transcription factor, as a promising target to control migration and invasion in breast cancer cells. Thus, an Ahr-based exploration was performed to identify a new Ahr agonist with inhibitory potentials on cancer cell motility...
2016: PloS One
https://www.readbyqxmd.com/read/27907057/the-effects-of-irreversible-electroporation-on-the-colon-in-a-porcine-model
#9
Xiaomei Luo, Xianjun Liang, Jiannan Li, Jian Shi, Wenlong Zhang, Wei Chai, Jiuping Wu, Shuai Guo, Gang Fang, Xulong Zhou, Jianhua Zhang, Kecheng Xu, Jianying Zeng, Lizhi Niu
BACKGROUND AND AIM: Irreversible electroporation (IRE) is a method of targeted cell ablation which has been suggested as a potential cancer therapy as it leaves structures such as blood vessels and the extracellular matrix intact, thereby allowing the rapid recovery of healthy tissue. Here, we investigated the effects of IRE on the colon in vivo in a porcine model. METHODS: IRE ablation was performed on the colon walls of 12 female Tibet mini-pigs, creating a total of 24 lesions...
2016: PloS One
https://www.readbyqxmd.com/read/27900346/preclinical-development-and-qualification-of-zfn-mediated-ccr5-disruption-in-human-hematopoietic-stem-progenitor-cells
#10
David L DiGiusto, Paula M Cannon, Michael C Holmes, Lijing Li, Anitha Rao, Jianbin Wang, Gary Lee, Philip D Gregory, Kenneth A Kim, Samuel B Hayward, Kathleen Meyer, Colin Exline, Evan Lopez, Jill Henley, Nancy Gonzalez, Victoria Bedell, Rodica Stan, John A Zaia
Gene therapy for HIV-1 infection is a promising alternative to lifelong combination antiviral drug treatment. Chemokine receptor 5 (CCR5) is the coreceptor required for R5-tropic HIV-1 infection of human cells. Deletion of CCR5 renders cells resistant to R5-tropic HIV-1 infection, and the potential for cure has been shown through allogeneic stem cell transplantation with naturally occurring homozygous deletion of CCR5 in donor hematopoietic stem/progenitor cells (HSPC). The requirement for HLA-matched HSPC bearing homozygous CCR5 deletions prohibits widespread application of this approach...
2016: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/27896368/targeting-of-the-wt191-138-fragment-to-human-dendritic-cells-improves-leukemia-specific-t-cell-responses-providing-an-alternative-approach-to-wt1-based-vaccination
#11
Nergui Dagvadorj, Anne Deuretzbacher, Daniela Weisenberger, Elke Baumeister, Johannes Trebing, Isabell Lang, Carolin Köchel, Markus Kapp, Kerstin Kapp, Andreas Beilhack, Thomas Hünig, Hermann Einsele, Harald Wajant, Götz Ulrich Grigoleit
Due to its immunogenicity and overexpression concomitant with leukemia progression, Wilms tumor protein 1 (WT1) is of particular interest for immunotherapy of AML relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT). So far, WT1-specific T-cell responses have mainly been induced by vaccination with peptides presented by certain HLA alleles. However, this approach is still not widely applicable in clinical practice due to common limitations of HLA restriction. Dendritic cell (DC) vaccines electroporated with mRNA encoding full-length protein have also been tested for generating WT1-derived peptides for presentation to T-cells...
November 28, 2016: Cancer Immunology, Immunotherapy: CII
https://www.readbyqxmd.com/read/27893744/optimization-of-irreversible-electroporation-protocols-for-in-vivo-myocardial-decellularization
#12
Yaniv Zager, David Kain, Natalie Landa, Jonathan Leor, Elad Maor
BACKGROUND: Irreversible electroporation (IRE) is a non-thermal cell ablation approach that induces selective damage to cell membranes only. The purpose of the current study was to evaluate and optimize its use for in-vivo myocardial decellularization. METHODS: Forty-two Sprague-Dawley rats were used to compare myocardial damage of seven different IRE protocols with anterior myocardial infarction damage. An in-vivo open thoracotomy model was used, with two-needle electrodes in the anterior ventricular wall...
2016: PloS One
https://www.readbyqxmd.com/read/27888407/targeting-the-rhesus-macaque-trim5%C3%AE-gene-to-enhance-the-susceptibility-of-cd4-t-cells-to-hiv-1-infection
#13
Xiaoli Wang, Qing Yu, Yahong Yuan, Zhiping Teng, Dongsheng Li, Yi Zeng
The host range of human immunodeficiency virus type 1 (HIV-1) is extremely narrow, which has hampered the establishment of non-human primate models for HIV-1 infection. The species-specific innate immune factor tripartite motif 5 alpha (TRIM5α) is a key molecule that confers potent resistance against HIV-1 infection. In this study, we targeted the TRIM5α gene of rhesus macaques (rhTRIM5α) using the transcription activator-like effector nuclease (TALEN) to study the effect on HIV-1 infection. CD4(+) T cells were separated from the peripheral blood of rhesus macaques by magnetic cell sorting, and the positive rate was greater than 99%...
November 25, 2016: Archives of Virology
https://www.readbyqxmd.com/read/27881406/enrichment-of-selective-mirnas-in-exosomes-and-delivery-of-exosomal-mirnas-in-vitro-and-in-vivo
#14
Duo Zhang, Heedoo Lee, Ziwen Zhu, Jasleen K Minhas, Yang Jin
Exosomes are nano vesicles secreted by cells, and contain various molecules including protein, lipid and DNA/RNA. They are crucial mediators of the intercellular communication and serve as promising vehicles for drug delivery and gene therapy. Recently, accumulating evidence suggest that microRNAs (miRNAs) may serve as new and potentially powerful targets for therapeutic interventions against various human diseases. However, steadily and effectively delivering miRNA mimics or inhibitors to target cells remains a major obstacle...
November 23, 2016: American Journal of Physiology. Lung Cellular and Molecular Physiology
https://www.readbyqxmd.com/read/27879339/mct8-deficiency-in-purkinje-cells-disrupts-embryonic-chicken-cerebellar-development
#15
Joke Delbaere, Pieter Vancamp, Stijn L J Van Herck, Nele M A Bourgeois, Mary J Green, Richard J T Wingate, Veerle M Darras
Inactivating mutations in the human SLC16A2 gene encoding the thyroid hormone transporter monocarboxylate transporter 8 (MCT8) result in the Allan-Herndon-Dudley syndrome accompanied by severe locomotor deficits. The underlying mechanisms of the associated cerebellar maldevelopment were studied using the chicken as a model. Electroporation of an MCT8-RNAi vector into the cerebellar anlage of a 3-day-old embryo allowed knockdown of MCT8 in Purkinje cell precursors. This resulted in downregulation of the thyroid hormone responsive gene RORα and the Purkinje cell specific differentiation marker LHX1/5 at day 6...
November 22, 2016: Journal of Endocrinology
https://www.readbyqxmd.com/read/27879219/induced-pluripotent-stem-cells-ipscs-derived-from-cerebrotendinous-xanthomatosis-ctx-patient-s-fibroblasts-carrying-a-r395s-mutation
#16
Philip Höflinger, Stefan Hauser, Yvonne Theurer, Stefanie Weißenberger, Carlo Wilke, Ludger Schöls
Induced pluripotent stem cells (iPSCs) were generated from dermal fibroblasts from a 60-year-old cerebrotendinous xanthomatosis (CTX) patient, carrying a homozygous mutation c. [1183C>A]; p. R395S in CYP27A1. Episomal plasmids encoding the pluripotency genes OCT4, SOX2, KLF4, L-MYC and LIN28 were introduced via electroporation. The generated line iPS-CTX-R395S has no sign of plasmid integration or chromosomal aberration and retained the mutation site in CYP27A1. Furthermore, iPSCs express pluripotency markers and are able to differentiate in all germ layers in vitro...
September 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27877916/study-of-local-intracellular-signals-regulating-axonal-morphogenesis-using-a-microfluidic-device
#17
Daiki Uryu, Tomohiro Tamaru, Azusa Suzuki, Rie Sakai, Yoshiyuki Konishi
The establishment and maintenance of axonal patterning is crucial for neuronal function. To identify the molecular systems that operate locally to control axonal structure, it is important to manipulate molecular functions in restricted subcellular areas for a long period of time. Microfluidic devices can be powerful tools for such purposes. In this study, we demonstrate the application of a microfluidic device to clarify the function of local Ca(2+) signals in axons. Membrane depolarization significantly induced axonal branch-extension in cultured cerebellar granule neurons (CGNs)...
2016: Science and Technology of Advanced Materials
https://www.readbyqxmd.com/read/27877159/efficacy-of-antibacterial-peptides-against-peptide-resistant-mrsa-is-restored-by-permeabilization-of-bacteria-membranes
#18
Joshua Ravensdale, Zachary Wong, Frances O'Brien, Keith Gregg
Clinical application of antimicrobial peptides (AMPs), as with conventional antibiotics, may be compromised by the development of bacterial resistance. This study investigated AMP resistance in methicillin resistant Staphylococcus aureus, including aspects related to the resilience of the resistant bacteria toward the peptides, the stability of resistance when selection pressures are removed, and whether resistance can be overcome by using the peptides with other membrane-permeabilising agents. Genotypically variant strains of S...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27874856/in-vivo-editing-of-the-human-mutant-rhodopsin-gene-by-electroporation-of-plasmid-based-crispr-cas9-in-the-mouse-retina
#19
Maria Carmela Latella, Maria Teresa Di Salvo, Fabienne Cocchiarella, Daniela Benati, Giulia Grisendi, Antonella Comitato, Valeria Marigo, Alessandra Recchia
The bacterial CRISPR/Cas system has proven to be an efficient tool for genetic manipulation in various organisms. Here we show the application of CRISPR-Cas9 technology to edit the human Rhodopsin (RHO) gene in a mouse model for autosomal dominant Retinitis Pigmentosa. We designed single or double sgRNAs to knock-down mutant RHO expression by targeting exon 1 of the RHO gene carrying the P23H dominant mutation. By delivering Cas9 and sgRNAs in a single plasmid we induced an efficient gene editing in vitro, in HeLa cells engineered to constitutively express the P23H mutant RHO allele...
November 22, 2016: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/27870918/time-dependent-impact-of-irreversible-electroporation-on-pancreas-liver-blood-vessels-and-nerves-a-systematic-review-of-experimental-studies
#20
J A Vogel, E van Veldhuisen, P Agnass, J Crezee, F Dijk, J Verheij, T M van Gulik, M R Meijerink, L G Vroomen, K P van Lienden, M G Besselink
INTRODUCTION: Irreversible electroporation (IRE) is a novel ablation technique in the treatment of unresectable cancer. The non-thermal mechanism is thought to cause mostly apoptosis compared to necrosis in thermal techniques. Both in experimental and clinical studies, a waiting time between ablation and tissue or imaging analysis to allow for cell death through apoptosis, is often reported. However, the dynamics of the IRE effect over time remain unknown. Therefore, this study aims to summarize these effects in relation to the time between treatment and evaluation...
2016: PloS One
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