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https://www.readbyqxmd.com/read/28640742/identification-of-amino-acids-within-norovirus-polymerase-involved-in-rna-binding-and-viral-replication
#1
Kang Rok Han, Bader Y Alhatlani, Sunyoung Cho, Ji-Hye Lee, Myra Hosmillo, Ian G Goodfellow, Kyung Hyun Kim, Jai Myung Yang
Until recently, molecular studies on human norovirus (HuNoV), a major causative agent of gastroenteritis, have been hampered by the lack of an efficient cell culture system. Murine norovirus-1 (MNV-1) has served as a surrogate model system for norovirus research, due to the availability of robust cell culture systems and reverse genetics. To identify amino acids involved in RNA synthesis by the viral RNA-dependent RNA polymerase (NS7), we constructed NS7 mutants in which basic amino acids surrounding the catalytic site were substituted with alanine...
June 22, 2017: Journal of General Virology
https://www.readbyqxmd.com/read/28627778/engineering-cho-cells-with-an-oncogenic-kit-improves-cells-growth-resilience-to-stress-and-productivity
#2
Mohamed Mahameed, Boaz Tirosh
An optimized biomanufacturing process in mammalian cells is contingent on the ability of the producing cells to reach high viable cell densities. In addition, at the peak of growth, cells need to continue producing the biological entity at a consistent quality. Thus, engineering cells with robust growth performance and resilience to variable stress conditions is highly desirable. The tyrosine kinase receptor, KIT, plays a key role in cell differentiation and the survival of several immune cell types. Its oncogenic mutant, D816V, endows cells with high proliferation capacity and resistance to kinase inhibitors...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28627725/proteomic-analysis-of-host-cell-protein-dynamics-in-the-supernatant-of-fc-fusion-protein-producing-cho-dg44-and-dukx-b11-cell-lines-in-batch-and-fed-batch-cultures
#3
Jin Hyoung Park, Jong Hwa Jin, In Jung Ji, Hyun Joo An, Jong Won Kim, Gyun Min Lee
Chinese hamster ovary (CHO) cells are the most widely used host cell lines for the commercial production of therapeutic proteins including Fc-fusion proteins. During the culture of recombinant CHO (rCHO) cells, host cell proteins (HCPs), secreted from viable cells and released from dead cells, accumulate extracellularly, potentially impairing product quality. In this study, the HCPs that accumulated extracellularly in batch and fed-batch cultures of Fc-fusion protein-producing rCHO cell lines (DG-Fc and DUKX-Fc) were identified and quantified using nanoflow liquid chromatography-tandem mass spectrometry (LC-MS/MS), followed by gene ontology and functional analysis...
June 19, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28625678/rapid-protein-production-from-stable-cho-cell-pools-using-plasmid-vector-and-the-cumate-gene-switch
#4
Adeline Poulain, Sylvie Perret, Félix Malenfant, Alaka Mullick, Bernard Massie, Yves Durocher
To rapidly produce large amounts of recombinant proteins, the generation of stable Chinese Hamster Ovary (CHO) cell pools represents a useful alternative to large-scale transient gene expression (TGE). We have developed a cell line (CHO(BRI/rcTA)) allowing the inducible expression of recombinant proteins, based on the cumate gene switch. After the identification of optimal plasmid DNA topology (supercoiled vs linearized plasmid) for PEIpro™ mediated transfection and of optimal conditions for methionine sulfoximine (MSX) selection, we were able to generate CHO(BRI/rcTA) pools producing high levels of recombinant proteins...
June 15, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28615417/t-cell-restricted-notch-signaling-contributes-to-pulmonary-th1-and-th2-immunity-during-cryptococcus-neoformans-infection
#5
Lori M Neal, Yafeng Qiu, Jooho Chung, Enze Xing, Woosung Cho, Antoni N Malachowski, Ashley R Sandy-Sloat, John J Osterholzer, Ivan Maillard, Michal A Olszewski
Cryptococcus neoformans is a ubiquitous, opportunistic fungal pathogen but the cell signaling pathways that drive T cell responses regulating antifungal immunity are incompletely understood. Notch is a key signaling pathway regulating T cell development, and differentiation and functional responses of mature T cells in the periphery. The targeting of Notch signaling within T cells has been proposed as a potential treatment for alloimmune and autoimmune disorders, but it is unknown whether disturbances to T cell immunity may render these patients vulnerable to fungal infections...
June 14, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28612559/-the-establishment-and-identification-of-stable-cho-cell-line-with-canine-transferrin-receptor-expression
#6
Wei Zhang, Yan Yan, Lu-Gen Bai, Yan-Ni Zhu, Ling Huang, Yu-Ning Sun
OBJECTIVES: To construct eukaryotic expressing recombinant vector of canine transferrin receptor gene (TfR ), then to transfect Chinese hamster ovary (CHO) cells with the vector for establishment of stable expression of TfR in CHO cell line. METHODS: The full-length TfR fragment was amplified by RT-PCR from the canine cells (walter reed dog cell, WRD) and then inserted into eukaryotic expression vector pCDNA3. After identification with enzyme digestion and sequencing, the recombinant vector was transfected into CHO cells by TransLipid Transfection Reagent...
January 2017: Sichuan da Xue Xue Bao. Yi Xue Ban, Journal of Sichuan University. Medical Science Edition
https://www.readbyqxmd.com/read/28611413/a-microengineered-model-of-rbc-transfusion-induced-pulmonary-vascular-injury
#7
Jeongyun Seo, David Conegliano, Megan Farrell, Minseon Cho, Xueting Ding, Thomas Seykora, Danielle Qing, Nilam S Mangalmurti, Dongeun Huh
Red blood cell (RBC) transfusion poses significant risks to critically ill patients by increasing their susceptibility to acute respiratory distress syndrome. While the underlying mechanisms of this life-threatening syndrome remain elusive, studies suggest that RBC-induced microvascular injury in the distal lung plays a central role in the development of lung injury following blood transfusion. Here we present a novel microengineering strategy to model and investigate this key disease process. Specifically, we created a microdevice for culturing primary human lung endothelial cells under physiological flow conditions to recapitulate the morphology and hemodynamic environment of the pulmonary microvascular endothelium in vivo...
June 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28601608/activation-of-unfolded-protein-response-pathway-is-important-for-valproic-acid-mediated-increase-in-immunoglobulin-g-productivity-in-recombinant-chinese-hamster-ovary-cells
#8
Kamal Prashad Segar, Vikas Chandrawanshi, Sarika Mehra
Process engineering to improve product quality and titers is gaining importance at late-stage cell culture process development. Valproic acid, a US Food and Drug Administration-approved histone deacetylase (HDAC) inhibitor, has been shown to improve cell culture performance with higher productivities and minimal effect on the product quality. However, the wider physiological impact of valproic acid on recombinant cells has not been investigated till date. In this study, we investigate the role of unfolded protein response pathway when immunoglobulin G (IgG)-secreting Chinese hamster ovary (CHO) cells are treated with valproic acid, resulting in a 3-fold increase in product titers and productivity...
June 7, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28599495/investigation-of-the-relative-biological-effectiveness-and-uniform-isobiological-killing-effects-of-irradiation-with-a-clinical-carbon-sobp-beam-on-dna-repair-deficient-cho-cells
#9
Shigeaki Sunada, Ian M Cartwright, Hirokazu Hirakawa, Akira Fujimori, Mitsuru Uesaka, Takamitsu A Kato
Spread-out Bragg peak (SOBP) C ions have been used clinically in charged particle radiation therapy for years. An SOBP beam consists of various monoenergetic Bragg peaks; however, the biological effect of irradiation with an SOBP beam track has not been well-studied. In order to determine the clinically prospective molecular targets, radiosensitivity to the beam track in DNA repair deficient cell lines was investigated. A total of four distinct Chinese hamster ovary (CHO) cell lines, including CHO10B2 (wild-type), V3 (protein kinase DNA-activated catalytic polypeptide deficient), 51D1 (RAD51 paralog D deficient) and PADR9 [poly(ADP-ribose) polymerase (PARP) deficient], were irradiated with gamma-rays, C ions (290 MeV/n) and Fe ions (500 MeV/n), in order to compare cellular lethality...
June 2017: Oncology Letters
https://www.readbyqxmd.com/read/28595074/protective-effect-of-arthrospira-platensis-extracts-against-ultraviolet-b-induced-cellular-senescence-through-inhibition-of-dna-damage-and-matrix-metalloproteinase-1-expression-in-human-dermal-fibroblasts
#10
Jeong-Ju Lee, Ki Bbeum Kim, Jina Heo, Dae-Hyun Cho, Hee-Sik Kim, Song Hee Han, Kyu Joong Ahn, In-Sook An, Sungkwan An, Seunghee Bae
Ultraviolet (UV) light exposure causes skin photoaging, which is known to be preventable and controllable by application of UV-protective agents. In this study, we demonstrated, for the first time, that the extract of microalgae Arthrospira platensis has a reverse effect on UV-induced photodamage such as loss of cell viability, cellular senescence, DNA damage, and collagen destruction in dermal fibroblasts. Forty-eight extracts were prepared from the cell biomass by controlling culture light conditions, extract solvents, and disruption methods...
May 31, 2017: Journal of Photochemistry and Photobiology. B, Biology
https://www.readbyqxmd.com/read/28586316/direct-3d-cell-printing-of-human-skin-with-functional-transwell-system
#11
Byoung Soo Kim, Jung-Seob Lee, Ge Gao, Dong-Woo Cho
Three-dimensional (3D) cell-printing has been emerging as a promising technology with which to build up human skin models by enabling rapid and versatile design. Despite the technological advances, challenges remain in the development of fully functional models that recapitulate complexities in the native tissue. Moreover, although several approaches have been explored for the development of biomimetic human skin models, the present skin models based on multistep fabrication methods using polydimethylsiloxane chips and commercial transwell inserts could be tackled by leveraging 3D cell-printing technology...
June 6, 2017: Biofabrication
https://www.readbyqxmd.com/read/28586020/altered-expression-of-epithelial-mesenchymal-transition-and-pluripotent-associated-markers-by-sex-steroid-hormones-in-human-embryonic-stem-cells
#12
So-Ye Jeon, Kyung-A Hwang, Cho-Won Kim, Eui-Bae Jeung, Kyung-Chul Choi
Embryonic stem (ES) cells are pluripotent stem cells derived from a developmental stage of pre‑implanted embryos. The present study investigated the effect of female sex steroid hormones on the characteristics of human ES cells by using a feeder‑free culture protocol. In a feeder‑free condition without sex hormones, human ES cells assumed the form of tightly packed cells that grow in a monolayer. The cells had clean and defined edges with no evidence of differentiation and expressed several markers specific for undifferentiated ES cells including POU class 5 homeobox 1 (POU5F1), sex determining region Y‑box 2 (SOX2) and NANOG homeobox (NANOG)...
July 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28585535/blood-brain-barrier-spheroids-as-an-in-vitro-screening-platform-for-brain-penetrating-agents
#13
Choi-Fong Cho, Justin M Wolfe, Colin M Fadzen, David Calligaris, Kalvis Hornburg, E Antonio Chiocca, Nathalie Y R Agar, Bradley L Pentelute, Sean E Lawler
Culture-based blood-brain barrier (BBB) models are crucial tools to enable rapid screening of brain-penetrating drugs. However, reproducibility of in vitro barrier properties and permeability remain as major challenges. Here, we report that self-assembling multicellular BBB spheroids display reproducible BBB features and functions. The spheroid core is comprised mainly of astrocytes, while brain endothelial cells and pericytes encase the surface, acting as a barrier that regulates transport of molecules. The spheroid surface exhibits high expression of tight junction proteins, VEGF-dependent permeability, efflux pump activity and receptor-mediated transcytosis of angiopep-2...
June 6, 2017: Nature Communications
https://www.readbyqxmd.com/read/28583763/measuring-clinically-relevant-endpoints-in-a-serum-free-three-dimensional-primary-cell-culture-system-of-human-osteoarthritic-articular-chondrocytes
#14
Grace Bundens, Andrea Buckley, LaBraya Milton, Kathryn Behling, Sarah Chmielewski, Ellen Cho, Xiomara Lozano-Torres, Abdulhafez Selim, Richard Lackman, Mindy George-Weinstein, Lawrence Miller, Marina D'Angelo
Osteoarthritis (OA) is characterized by degeneration of articular cartilage within the joint, inflammation and pain. The purpose of this study was to develop a primary, serum free cell culture system of human osteoarthritic articular chondrocytes (HOACs) with which to study manifestations of the disease process. Joint tissues were obtained from OA patients undergoing total knee arthroplasty (TKA). HOACs isolated from the femoral condyles and tibial plateau of the same side were combined, plated in three-dimensional, alginate beads and cultured for five days in serum, hormone and protein free medium...
June 2, 2017: Experimental Cell Research
https://www.readbyqxmd.com/read/28583554/enterobacter-cloacae-related-necrotizing-fasciitis-after-peritoneal-dialysis-in-delayed-graft-function-a-case-report
#15
Y-M Choe, K-M Park, Y S Jeon, S G Cho, K C Hong, W Y Shin, J H Song
INTRODUCTION: Necrotizing fasciitis is an uncommon soft-tissue infection that involves the superficial fascia, subcutaneous fat, and deep fascia. Herein, we report the first case of Enterobacter cloacae-related necrotizing fasciitis after peritoneal dialysis in delayed graft function. CASE: A 58-year-old man, who was a hepatitis B-viral carrier and had atrial fibrillation, received cadaveric renal transplantation with peritoneal dialysis and encountered delayed graft function...
June 2017: Transplantation Proceedings
https://www.readbyqxmd.com/read/28577512/possible-radioprotective-effect-of-folic-acid-supplementation-on-low-dose-ionizing-radiation-induced-genomic-instability-in-vitro
#16
Gisel Padula, María Virginia Ponzinibbio, Analia I Seoane
Ionizing radiation (IR) induces DNA damage through production of single and double-strand breaks and reactive oxygen species (ROS). Folic acid (FA) prevents radiation-induced DNA damage by modification of DNA synthesis and/or repair and as a radical scavenger. We hypothesized that in vitro supplementation with FA will decrease the sensitivity of cells to genetic damage induced by low dose of ionizing radiation. Annexin V, comet and micronucleus assays were performed in cultured CHO cells. After 7 days of pre-treatment with 0, 100, 200 or 300 nM FA, cultures were exposed to radiation (100 mSv)...
August 2016: Indian Journal of Experimental Biology
https://www.readbyqxmd.com/read/28570275/long-term-culture-of-human-liver-tissue-with-advanced-hepatic-functions
#17
Soon Seng Ng, Anming Xiong, Khanh Nguyen, Marilyn Masek, Da Yoon No, Menashe Elazar, Eyal Shteyer, Mark A Winters, Amy Voedisch, Kate Shaw, Sheikh Tamir Rashid, Curtis W Frank, Nam Joon Cho, Jeffrey S Glenn
A major challenge for studying authentic liver cell function and cell replacement therapies is that primary human hepatocytes rapidly lose their advanced function in conventional, 2-dimensional culture platforms. Here, we describe the fabrication of 3-dimensional hexagonally arrayed lobular human liver tissues inspired by the liver's natural architecture. The engineered liver tissues exhibit key features of advanced differentiation, such as human-specific cytochrome P450-mediated drug metabolism and the ability to support efficient infection with patient-derived inoculums of hepatitis C virus...
June 2, 2017: JCI Insight
https://www.readbyqxmd.com/read/28566555/cross-linked-matrix-rigidity-and-soluble-retinoids-synergize-in-nuclear-lamina-regulation-of-stem-cell-differentiation
#18
Irena L Ivanovska, Joe Swift, Kyle Spinler, Dave Dingal, Sangkyun Cho, Dennis E Discher
Synergistic cues from extracellular matrix and soluble factors are often obscure in differentiation. Here, the rigidity of crosslinked collagen synergizes with retinoids in the osteogenesis of human marrow mesenchymal stem cells (MSCs). Collagen nano-films serve as a model matrix that MSCs can easily deform unless the film is enzymatically crosslinked, which promotes the spreading of cells and the stiffening of nuclei as both actomyosin assembly and nucleoskeletal lamin-A increase. Expression of lamin-A is known to be controlled by Retinoic Acid Receptor (RAR) transcription factors, but rigid matrix is needed to induce rapid nuclear accumulation of the RARG isoform and subsequent expression of osteogenic markers...
May 31, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28565035/clinical-experience-with-wbc-pet-ct-in-adpkd-patients-with-suspected-cyst-infection-a-prospective-case-series
#19
Hyunsuk Kim, Yun Kyu Oh, Hayne Cho Park, Seokwoo Park, Soojin Lee, Ho-Young Lee, Young-Hwan Hwang, Curie Ahn
AIMS: Cyst infection (CI) is a common problem in patients with autosomal dominant polycystic kidney disease (ADPKD). Localization is of great importance in CI. We describe the clinical experience with [18F] FDG-labeled white-blood cell (WBC) PET/CT in detecting CI in ADPKD. METHODS: 19 ADPKD patients (M:F = 7:12) suspected of having CI were enrolled in this prospective study. All underwent WBC-PET/CT and MRI or CT. The degree of their WBC accumulation was evaluated from the maximal standardized uptake value of cystic wall...
May 31, 2017: Nephrology
https://www.readbyqxmd.com/read/28557288/the-ef-1%C3%AE-promoter-maintains-high-level-transgene-expression-from-episomal-vectors-in-transfected-cho-k1-cells
#20
Xiaoyin Wang, Chongjie Xu, Zhengwei Tian, Xi Zhang, Danhua Xu, Qin Li, Junhe Zhang, Tianyun Wang
In our previous study, we demonstrated that episomal vectors based on the characteristic sequence of matrix attachment regions (MARs) and containing the cytomegalovirus (CMV) promoter allow transgenes to be maintained episomally in Chinese hamster ovary (CHO) cells. However, the transgene expression was unstable and the number of copies was low. In this study, we focused on enhancers, various promoters and promoter variants that could improve the transgene expression stability, expression magnitude (level) and the copy number of a MAR-based episomal vector in CHO-K1 cells...
May 30, 2017: Journal of Cellular and Molecular Medicine
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