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https://www.readbyqxmd.com/read/29159981/understanding-the-metabolic-burden-of-recombinant-antibody-production-in-saccharomyces-cerevisiae-using-a-quantitative-metabolomics-approach
#1
Jorg C de Ruijter, Essi V Koskela, Jatin Nandania, Alexander D Frey, Vidya Velagapudi
The cellular changes induced by heterologous protein expression in the yeast Saccharomyces cerevisiae have been analyzed on many levels and found to be significant. However, even though high-level protein production poses a metabolic burden, evaluation of the expression host at the level of the metabolome has often been neglected. We present a comparison of metabolite profiles of a wild-type strain with those of three strains producing recombinant antibody variants of increasing size and complexity: a scFv fragment, a scFv-Fc fusion protein, and a full-length IgG molecule...
November 20, 2017: Yeast
https://www.readbyqxmd.com/read/29159802/adenovirus-vector-based-prime-boost-vaccination-via-heterologous-routes-induces-cervicovaginal-cd8-t-cell-responses-against-hpv16-oncoproteins
#2
Nicolas Çuburu, Selina Khan, Cynthia D Thompson, Rina Kim, Jort Vellinga, Roland Zahn, Douglas R Lowy, Gert Scheper, John T Schiller
Recent advances in immunotherapy against cancer underscore the importance of T lymphocytes and tumor microenvironment, but few vaccines targeting cancer have been approved likely due in part to the dearth of common tumor antigens, insufficient immunogenicity and the evolution of immune evasion mechanisms during the progression to malignancy. Human papillomaviruses (HPV) are the primary etiologic agents of cervical cancer and progression from persistent HPV-infection to cervical intraepithelial lesions and eventually cancer requires persistent expression of the oncoproteins E6 and E7...
November 21, 2017: International Journal of Cancer. Journal International du Cancer
https://www.readbyqxmd.com/read/29158520/myd88-promotes-myoblast-fusion-in-a-cell-autonomous-manner
#3
Sajedah M Hindi, Jonghyun Shin, Yann S Gallot, Alex R Straughn, Adriana Simionescu-Bankston, Lubna Hindi, Guangyan Xiong, Robert P Friedland, Ashok Kumar
Myoblast fusion is an indispensable step for skeletal muscle development, postnatal growth, and regeneration. Myeloid differentiation primary response gene 88 (MyD88) is an adaptor protein that mediates Toll-like receptors and interleukin-1 receptor signaling. Here we report a cell-autonomous role of MyD88 in the regulation of myoblast fusion. MyD88 protein levels are increased during in vitro myogenesis and in conditions that promote skeletal muscle growth in vivo. Deletion of MyD88 impairs fusion of myoblasts without affecting their survival, proliferation, or differentiation...
November 20, 2017: Nature Communications
https://www.readbyqxmd.com/read/29158488/annexin-a4-and-a6-induce-membrane-curvature-and-constriction-during-cell-membrane-repair
#4
Theresa Louise Boye, Kenji Maeda, Weria Pezeshkian, Stine Lauritzen Sønder, Swantje Christin Haeger, Volker Gerke, Adam Cohen Simonsen, Jesper Nylandsted
Efficient cell membrane repair mechanisms are essential for maintaining membrane integrity and thus for cell life. Here we show that the Ca(2+)- and phospholipid-binding proteins annexin A4 and A6 are involved in plasma membrane repair and needed for rapid closure of micron-size holes. We demonstrate that annexin A4 binds to artificial membranes and generates curvature force initiated from free edges, whereas annexin A6 induces constriction force. In cells, plasma membrane injury and Ca(2+) influx recruit annexin A4 to the vicinity of membrane wound edges where its homo-trimerization leads to membrane curvature near the edges...
November 20, 2017: Nature Communications
https://www.readbyqxmd.com/read/29158476/programmable-dna-looping-using-engineered-bivalent-dcas9-complexes
#5
Nan Hao, Keith E Shearwin, Ian B Dodd
DNA looping is a ubiquitous and critical feature of gene regulation. Although DNA looping can be efficiently detected, tools to readily manipulate DNA looping are limited. Here we develop CRISPR-based DNA looping reagents for creation of programmable DNA loops. Cleavage-defective Cas9 proteins of different specificity are linked by heterodimerization or translational fusion to create bivalent complexes able to link two separate DNA regions. After model-directed optimization, the reagents are validated using a quantitative DNA looping assay in E...
November 20, 2017: Nature Communications
https://www.readbyqxmd.com/read/29158362/cd38-bispecific-antibody-pretargeted-radioimmunotherapy-for-multiple-myeloma-and-other-b-cell-malignancies
#6
Damian J Green, Shyril O'Steen, Yukang Lin, Melilssa L Comstock, Aimee L Kenoyer, Donald K Hamlin, D Scott Wilbur, Darrell R Fisher, Margaret Nartea, Mark D Hylarides, Ajay K Gopal, Theodore A Gooley, Johnnie J Orozco, Brian G Till, Kelly D Orcutt, K Dane Wittrup, Oliver W Press
Pretargeted radioimmunotherapy (PRIT) has demonstrated remarkable efficacy targeting tumor antigens, but immunogenicity and endogenous biotin blocking may limit clinical translation. We describe a new PRIT approach for the treatment of Multiple Myeloma (MM) and other B cell malignancies, for which we developed an anti-CD38 bispecific fusion protein that eliminates endogenous biotin interference and immunogenic elements. In murine xenograft models of MM and non-Hodgkin lymphoma (NHL), the CD38 bispecific construct demonstrated excellent blood clearance and tumor targeting...
November 20, 2017: Blood
https://www.readbyqxmd.com/read/29158309/why-the-need-and-how-to-approach-the-functional-diversity-of-extracellular-vesicles
#7
REVIEW
Mercedes Tkach, Joanna Kowal, Clotilde Théry
In the past decade, cell-to-cell communication mediated by exosomes has attracted growing attention from biomedical scientists and physicians, leading to several recent publications in top-tier journals. Exosomes are generally defined as secreted membrane vesicles, or extracellular vesicles (EVs), corresponding to the intraluminal vesicles of late endosomal compartments, which are secreted upon fusion of multi-vesicular endosomes with the cell's plasma membrane. Cells, however, were shown to release other types of EVs, for instance, by direct budding off their plasma membrane...
January 5, 2018: Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
https://www.readbyqxmd.com/read/29158139/expression-and-evaluation-of-recombinant-p32-protein-based-elisa-for-sero-diagnostic-potential-of-capripox-in-sheep-and-goats
#8
Gnanavel Venkatesan, Mahesh Kumar Teli, Muthu Sankar, Amit Kumar, M Dashprakash, Sargam Arya, Aparna Madhavan, Muthannan Andavar Ramakrisnan, Awadh Bihari Pandey
The study is aimed to develop and evaluate a recombinant P32 protein based ELISA for sero-monitoring and sero-surveillance using known and suspected serum samples for capripox from sheep and goats. Truncated P32 gene of goatpox virus (with an ORF of 750 bp) was expressed in E. coli BL-21 CodonPlus (DE3) cells using pET32a vector and characterized by SDS-PAGE analysis and confirmed by western blotting as 48 KDa Poly-His tagged fusion protein. The protein was purified under denaturing conditions using 8M urea and characterized by SDS-PAGE and immunoblotting...
November 17, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/29156757/a-pten-col17a1-fusion-gene-and-its-novel-regulatory-role-in-collagen-xvii-expression-and-gbm-malignance
#9
Xiaoyan Yan, Chuanbao Zhang, Tingyu Liang, Fan Yang, Haoyuan Wang, Fan Wu, Wen Wang, Zheng Wang, Wen Cheng, Jiangnan Xu, Tao Jiang, Jing Chen, Yaozhong Ding
Collagen XVII expression has recently been demonstrated to be correlated with the tumor malignance. While Collagen XVII is known to be widely distributed in neurons of the human brain, its precise role in pathogenesis of glioblastoma multiforme (GBM) is unknown. In this study, we identified and characterized a new PTEN-COL17A1 fusion gene in GMB using transcriptome sequencing. Although fusion gene did not result in measurable fusion protein production, its presence is accompanied with high levels of COL17A1 expression, revealed a novel regulatory mechanism of Collagen XVII expression by PTEN-COL17A1 gene fusion...
October 17, 2017: Oncotarget
https://www.readbyqxmd.com/read/29156603/adding-an-artificial-tail-anchor-to-a-peptide-based-hiv-1-fusion-inhibitor-for-improvement-of-its-potency-and-resistance-profile
#10
Shan Su, Zhenxuan Ma, Chen Hua, Weihua Li, Lu Lu, Shibo Jiang
Peptides derived from the C-terminal heptad repeat (CHR) of human immunodeficiency virus type 1 (HIV-1) envelope protein transmembrane subunit gp41, such as T20 (enfuvirtide), can bind to the N-terminal heptad repeat (NHR) of gp41 and block six-helix bundle (6-HB) formation, thus inhibiting HIV-1 fusion with the target cell. However, clinical application of T20 is limited because of its low potency and genetic barrier to resistance. HP23, the shortest CHR peptide, exhibits better anti-HIV-1 activity than T20, but the HIV-1 strains with E49K mutations in gp41 will become resistant to it...
November 20, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/29156538/combined-effects-of-simulated-microgravity-and-radiation-exposure-on-osteoclast-cell-fusion
#11
Srinivasan Shanmugarajan, Ye Zhang, Maria Moreno-Villanueva, Ryan Clanton, Larry H Rohde, Govindarajan T Ramesh, Jean D Sibonga, Honglu Wu
The loss of bone mass and alteration in bone physiology during space flight are one of the major health risks for astronauts. Although the lack of weight bearing in microgravity is considered a risk factor for bone loss and possible osteoporosis, organisms living in space are also exposed to cosmic radiation and other environmental stress factors. As such, it is still unclear as to whether and by how much radiation exposure contributes to bone loss during space travel, and whether the effects of microgravity and radiation exposure are additive or synergistic...
November 18, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29155724/generation-of-a-gene-disrupted-streptococcus-mutans-strain-without-gene-cloning
#12
Takatoshi Murata, Ayako Okada, Khairul Matin, Nobuhiro Hanada
Typical methods for the elucidation of the function of a particular gene involve comparative phenotypic analyses of the wild-type strain and a strain in which the gene of interest has been disrupted. A gene-disruption DNA construct containing a suitable antibiotic resistance marker gene is useful for the generation of gene-disrupted strains in bacteria. However, conventional construction methods, which require gene cloning steps, involve complex and time-consuming protocols. Here, a relatively facile, rapid, and cost-effective method for targeted gene disruption in Streptococcus mutans is described...
October 23, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29154869/evaluating-different-dna-binding-domains-to-modulate-l1-orf2p-driven-site-specific-retrotransposition-events-in-human-cells
#13
Catherine M Ade, Rebecca S Derbes, Bradley J Wagstaff, Sara B Linker, Travis B White, Dawn Deharo, Victoria P Belancio, Zoltán Ivics, Astrid M Roy-Engel
DNA binding domains (DBDs) have been used with great success to impart targeting capabilities to a variety of proteins creating highly useful genomic tools. We evaluated the ability of five types of DBDs and strategies (AAV Rep proteins, Cre, TAL effectors, zinc finger proteins, and Cas9/gRNA system) to target the L1 ORF2 protein to drive retrotransposition of Alu inserts to specific sequences in the human genome. First, we find that the L1 ORF2 protein tolerates the addition of protein domains both at the amino- and carboxy-terminus...
November 14, 2017: Gene
https://www.readbyqxmd.com/read/29153777/vaccination-with-a-human-parainfluenza-virus-type-3-chimeric-fhn-glycoprotein-formulated-with-a-combination-adjuvant-induces-protective-immunity
#14
R Garg, R Brownlie, L Latimer, V Gerdts, A Potter, S van Drunen Littel-van den Hurk
Human parainfluenza virus type 3 (PIV3) is a major cause of lower respiratory disease i.e. bronchitis, bronchiolitis or pneumonia, in infants and young children. Presently there is no licensed vaccine against PIV3. To produce an effective subunit vaccine, a chimeric FHN glycoprotein consisting of the N-terminal ectodomain of the fusion (F) protein linked to the haemagglutinin-neuraminidase (HN) protein without transmembrane domain, and secreted forms of the individual F and HN glycoproteins, were expressed in mammalian cells and purified...
November 16, 2017: Vaccine
https://www.readbyqxmd.com/read/29152418/generation-and-characterization-of-cross-neutralizing-human-monoclonal-antibody-against-4-serotypes-of-dengue-virus-without-enhancing-activity
#15
Subenya Injampa, Nataya Muenngern, Chonlatip Pipattanaboon, Surachet Benjathummarak, Khwanchit Boonha, Hathairad Hananantachai, Waranya Wongwit, Pongrama Ramasoota, Pannamthip Pitaksajjakul
Background: Dengue disease is a leading cause of illness and death in the tropics and subtropics. Most severe cases occur among patients secondarily infected with a different dengue virus (DENV) serotype compared with that from the first infection, resulting in antibody-dependent enhancement activity (ADE). Our previous study generated the neutralizing human monoclonal antibody, D23-1B3B9 (B3B9), targeting the first domain II of E protein, which showed strong neutralizing activity (NT) against all four DENV serotypes...
2017: PeerJ
https://www.readbyqxmd.com/read/29152155/chibby-1-a-new-component-of-%C3%AE-catenin-signaling-in-chronic-myeloid-leukemia
#16
REVIEW
Manuela Mancini, Simona Soverini, Gabriele Gugliotta, Maria Alessandra Santucci, Gianantonio Rosti, Michele Cavo, Giovanni Martinelli, Fausto Castagnetti
Chibby 1 (CBY1) is a small and evolutionarily conserved protein, which act as β-catenin antagonist. CBY1 is encoded by C22orf2 (22q13.1) Its antagonistic function on β-catenin involves the direct interaction with: The C-terminal activation domain of β-catenin, which hinders β-catenin binding with Tcf/Lef transcription factors hence repressing β-catenin transcriptional activation. 14-3-3 scaffolding proteins (σ or ξ), which drive CBY1 nuclear export into a stable tripartite complex with β-catenin. The relative proximity of C22orf2 gene encoding for CBY1 to the BCR breakpoint on chromosome 22q11, whose translocation and rearrangement with the c-ABL is the causative event of chronic myeloid leukemia (CML), suggested that gene haploinsufficiency may play a role in the disease pathogenesis and progression...
October 20, 2017: Oncotarget
https://www.readbyqxmd.com/read/29152069/the-transcriptional-coregulator-nab2-is-a-target-gene-for-the-wilms-tumor-gene-1-protein-wt1-in-leukemic-cells
#17
Helena Jernmark Nilsson, Giorgia Montano, Tove Ullmark, Andreas Lennartsson, Kristina Drott, Linnea Järvstråt, Björn Nilsson, Karina Vidovic, Urban Gullberg
The Wilms' tumor gene 1 (WT1) is recurrently mutated in acute myeloid leukemia. Mutations and high expression of WT1 associate with a poor prognosis. In mice, WT1 cooperates with the RUNX1/RUNX1T1 (AML1/ETO) fusion gene in the induction of acute leukemia, further emphasizing a role for WT1 in leukemia development. Molecular mechanisms for WT1 are, however, incompletely understood. Here, we identify the transcriptional coregulator NAB2 as a target gene of WT1. Analysis of gene expression profiles of leukemic samples revealed a positive correlation between the expression of WT1 and NAB2, as well as a non-zero partial correlation...
October 20, 2017: Oncotarget
https://www.readbyqxmd.com/read/29152063/acquisition-of-an-oncogenic-fusion-protein-is-sufficient-to-globally-alter-the-landscape-of-mirna-expression-to-inhibit-myogenic-differentiation
#18
Jacob M Loupe, Patrick J Miller, Judy S Crabtree, Jovanny Zabaleta, Andrew D Hollenbach
The differentiation status of tumors is used as a prognostic indicator, with tumors comprised of less differentiated cells exhibiting higher levels of aggressiveness that correlate with a poor prognosis. Although oncogenes contribute to blocking differentiation, it is not clear how they globally alter miRNA expression during differentiation to achieve this result. The pediatric sarcoma Alveolar Rhabdomyosarcoma, which is primarily characterized by the expression of the PAX3-FOXO1 oncogenic fusion protein, consists of undifferentiated muscle cells...
October 20, 2017: Oncotarget
https://www.readbyqxmd.com/read/29152060/inhibition-of-chk1-sensitizes-ewing-sarcoma-cells-to-the-ribonucleotide-reductase-inhibitor-gemcitabine
#19
Kelli L Goss, Stacia L Koppenhafer, Kathryn M Harmoney, William W Terry, David J Gordon
Ewing sarcoma is a bone and soft tissue sarcoma that occurs in children and young adults. The EWS-FLI1 gene fusion is the driver mutation in most Ewing sarcoma tumors and functions, in part, as an aberrant transcription factor. We recently identified that Ewing sarcoma cells are sensitive to inhibition of ribonucleotide reductase (RNR), which catalyzes the formation of deoxyribonucleotides from ribonucleotides. In this report, we show that Ewing sarcoma cells are sensitive to treatment with clofarabine, which is a nucleoside analogue and allosteric inhibitor of RNR...
October 20, 2017: Oncotarget
https://www.readbyqxmd.com/read/29151902/coexistence-of-p210-bcr-abl-and-cbf%C3%AE-myh11-fusion-genes-in-myeloid-leukemia-a-report-of-4-cases
#20
Yuan-Yuan Wang, Wen-Jing Ding, Feng Jiang, Zi-Xing Chen, Jian-Nong Cen, Xiao-Fei Qi, Jian-Ying Liang, Dan-Dan Liu, Jin-Lan Pan, Su-Ning Chen
Numerous acquired molecular and cytogenetic abnormalities are strongly associated with hematological malignancies. The breakpoint cluster region-ABL proto-oncogene 1 (BCR-ABL) rearrangement leads to a p210 chimeric protein in typical chronic myeloid leukemia (CML), whereas 17-25% of patients with acute lymphocytic leukemia and 0.9-3% patients with de novo acute myeloid leukemia (AML) carry a p190(BCR-ABL) fusion protein. Cases of patients with AML/CML carrying two specific primary molecular changes, BCR-ABL and core binding factor-β-myosin heavy chain 11 (CBFβ-MYH11) fusion genes have been rarely reported...
November 2017: Oncology Letters
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