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https://www.readbyqxmd.com/read/28435224/investigation-of-triamterene-as-an-inhibitor-of-the-tgr5-receptor-identification-in-cells-and-animals
#1
Yingxiao Li, Kai Chun Cheng, Chiang-Shan Niu, Shih-Hsiang Lo, Juei-Tang Cheng, Ho-Shan Niu
BACKGROUND: G-protein-coupled bile acid receptor 1 (GPBAR1, also known as TGR5) has been shown to participate in glucose homeostasis. In animal models, a TGR5 agonist increases incretin secretion to reduce hyperglycemia. Many agonists have been developed for clinical use. However, the effects of TGR5 blockade have not been studied extensively, with the exception of studies using TGR5 knockout mice. Therefore, we investigated the potential effect of triamterene on TGR5. METHODS: We transfected the TGR5 gene into cultured Chinese hamster ovary cells (CHO-K1 cells) to express TGR5...
2017: Drug Design, Development and Therapy
https://www.readbyqxmd.com/read/28432429/enhanced-production-of-recombinant-proteins-by-a-small-molecule-protein-synthesis-enhancer-in-combination-with-an-antioxidant-in-recombinant-chinese-hamster-ovary-cells
#2
Joseph Camire, Dongjoo Kim, Soonjo Kwon
The improvement in the production of recombinant proteins has been linked in a number of small molecules such as carboxylic acids to the inhibition of histone deacetylase, leading to increased transcription of genes. However, carboxylic acids such as pentanoic acid and butanoic acid have been shown to promote an apoptotic response in Chinese hamster ovary (CHO) cell culture. Supplementation of cultures with antioxidants has shown the ability to reduce the apoptotic response of carboxylic acid supplementation, leading to increased therapeutic protein production...
April 21, 2017: Bioprocess and Biosystems Engineering
https://www.readbyqxmd.com/read/28419922/cadmium-induces-ca-2-mediated-calpain-1-caspase-3-dependent-apoptosis-in-primary-cultured-rat-proximal-tubular-cells
#3
Hong Wang, Nianhui Zhai, Ying Chen, Haibin Xu, Kehe Huang
Calcium, as a ubiquitous second messenger, governs a large array of cellular processes and is necessary for cell survival. More recently, it was observed that the cytosolic Ca(2+) concentration ([Ca(2+)]c) elevation could induce apoptosis in primary cultured rat proximal tubular (rPT) cells exposed to cadmium (Cd), but the concrete mechanism is still unclear. This study was designed to investigate the signal pathway involved in [Ca(2+)]c elevation-mediated apoptosis. The results confirmed the elevation of [Ca(2+)]c by confocal microscopy and enhancement of the apoptosis by Hoechst 33258 staining and flow cytometer when rPT cells were exposed to Cd for 12h...
April 8, 2017: Journal of Inorganic Biochemistry
https://www.readbyqxmd.com/read/28414024/novel-high-throughput-cell-based-hybridoma-screening-methodology-using-the-celigo-image-cytometer
#4
Haohai Zhang, Leo Li-Ying Chan, William Rice, Nasim Kassam, Maria Serena Longhi, Haitao Zhao, Simon C Robson, Wenda Gao, Yan Wu
Hybridoma screening is a critical step for antibody discovery, which necessitates prompt identification of potential clones from hundreds to thousands of hybridoma cultures against the desired immunogen. Technical issues associated with ELISA- and flow cytometry-based screening limit accuracy and diminish high-throughput capability, increasing time and cost. Conventional ELISA screening with coated antigen is also impractical for difficult-to-express hydrophobic membrane antigens or multi-chain protein complexes...
April 14, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28410190/dietary-oleuropein-inhibits-tumor-angiogenesis-and-lymphangiogenesis-in-the-b16f10-melanoma-allograft-model-a-mechanism-for-the-suppression-of-high-fat-diet-induced-solid-tumor-growth-and-lymph-node-metastasis
#5
Hyerim Song, Do Young Lim, Jae In Jung, Han Jin Cho, So Young Park, Gyoo Taik Kwon, Young-Hee Kang, Ki Won Lee, Myung-Sook Choi, Jung Han Yoon Park
Previously, we reported that high-fat-diet (HFD)-induced obesity stimulates melanoma progression in the B16F10 allograft model. In this study, we examined whether oleuropein (OL), the most abundant phenolic compound in olives, inhibits HFD-induced melanoma progression. Four-week-old male C57BL/6N mice were fed a HFD-diet with or without OL. After 16 weeks of feeding, B16F10-luc cells were subcutaneously injected and the primary tumor was resected 3 weeks later. OL suppressed HFD-induced solid tumor growth. In the tumor tissues, OL reduced HFD-induced expression of angiogenesis (CD31, VE-cadherin, VEGF-A, and VEGFR2), lymphangiogenesis (LYVE-1, VEGF-C, VEGF-D, and VEGFR3), and hypoxia (HIF-1α and GLUT-1) markers as well as HFD-induced increases in lipid vacuoles and M2 macrophages (MΦs)...
March 31, 2017: Oncotarget
https://www.readbyqxmd.com/read/28409838/modulation-and-modeling-of-monoclonal-antibody-n-linked-glycosylation-in-mammalian-cell-perfusion-reactors
#6
Daniel J Karst, Ernesto Scibona, Elisa Serra, Jean-Marc Bielser, Jonathan Souquet, Matthieu Stettler, Hervé Broly, Miroslav Soos, Massimo Morbidelli, Thomas K Villiger
Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity...
April 14, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28409824/a-comparative-genomic-hybridization-approach-to-study-gene-copy-number-variations-among-chinese-hamster-cell-lines
#7
Nandita Vishwanathan, Arpan Bandyopadhyay, Hsu-Yuan Fu, Kathryn C Johnson, Nathan M Springer, Wei-Shou Hu
Chinese Hamster Ovary (CHO) cells are aneuploid in nature. The genome of recombinant protein producing CHO cell lines continuously undergoes changes in its structure and organization. We analyzed nine cell lines, including parental cell lines, using a comparative genomic hybridization (CGH) array focused on gene-containing regions. The comparison of CGH with copy-number estimates from sequencing data showed good correlation. Hierarchical clustering of the gene copy number variation data from CGH data revealed the lineage relationships between the cell lines...
April 14, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28409820/identification-and-control-of-novel-growth-inhibitors-in-fed-batch-cultures-of-chinese-hamster-ovary-cells
#8
Bhanu Chandra Mulukutla, Jaitashree Kale, Taylor Kalomeris, Michaela Jacobs, Gregory W Hiller
Chinese hamster ovary (CHO) cells in culture are known to consume large amounts of nutrients and divert most of them towards byproducts, some of which, including lactate and ammonia, are known to be toxic in nature. Glucose limitation strategies can successfully control lactate accumulation in fed-batch cultures yielding higher peak cell densities and titers. Interestingly, even in such optimized cultures, cell growth slows and eventually stops, indicating the emergence of other factors that negatively affect cell growth...
April 14, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28394476/enhanced-self-renewal-and-accelerated-differentiation-of-human-fetal-neural-stem-cells-using-graphene-oxide-nanoparticles
#9
Jin Kim, Kisuk Yang, Jong Seung Lee, Yong Hwa Hwang, Hyun-Ji Park, Kook In Park, Dong Yun Lee, Seung-Woo Cho
Graphene oxide (GO) has received increasing attention in bioengineering fields due to its unique biophysical and electrical properties, along with excellent biocompatibility. The application of GO nanoparticles (GO-NPs) to engineer self-renewal and differentiation of human fetal neural stem cells (hfNSCs) is reported. GO-NPs added to hfNSC culture during neurosphere formation substantially promote cell-to-cell and cell-to-matrix interactions in neurospheres. Accordingly, GO-NP-treated hfNSCs show enhanced self-renewal ability and accelerated differentiation compared to untreated cells, indicating the utility of GO in developing stem cell therapies for neurogenesis...
April 10, 2017: Macromolecular Bioscience
https://www.readbyqxmd.com/read/28393020/effects-of-exogenous-recombinant-human-bone-morphogenic-protein-7-on-the-corneal-epithelial-mesenchymal-transition-and-fibrosis
#10
Jin Kwon Chung, Shin Ae Park, Hee Sun Hwang, Kwang Sung Kim, Yang Je Cho, Yong Sung You, Young Sik Kim, Ju Woong Jang, Sung Jin Lee
AIM: To evaluate the effect of exogenous recombinant human bone morphogenic protein-7 (rhBMP-7) on transforming growth factor-β (TGF-β)-induced epithelial mesenchymal cell transition (EMT) and assessed its antifibrotic effect via topical application. METHODS: The cytotoxic effect of rhBMP-7 was evaluated and the EMT of human corneal epithelial cells (HECEs) was induced by TGF-β. HECEs were then cultured in the presence of rhBMP-7 and/or hyaluronic acid (HA). EMT markers, fibronectin, E-cadherin, α-smooth muscle actin (α-SMA), and matrix metaloproteinase-9 (MMP-9), were evaluated...
2017: International Journal of Ophthalmology
https://www.readbyqxmd.com/read/28390866/activation-of-transforming-growth-factor-beta-1-signaling-in-gastric-cancer-associated-fibroblasts-increases-their-motility-via-expression-of-rhomboid-5-homolog-2-and-ability-to-induce-invasiveness-of-gastric-cancer-cells
#11
Takatsugu Ishimoto, Keisuke Miyake, Tannistha Nandi, Masakazu Yashiro, Nobuyuki Onishi, Kie Kyon Huang, Suling Joyce Lin, Ramnarayanan Kalpana, Su Ting Tay, Yuka Suzuki, Byoung Chul Cho, Daisuke Kuroda, Kota Arima, Daisuke Izumi, Masaaki Iwatsuki, Yoshifumi Baba, Eiji Oki, Masayuki Watanabe, Hideyuki Saya, Kosei Hirakawa, Hideo Baba, Patrick Tan
BACKGROUND & AIMS: Fibroblasts that interact with cancer cells are called cancer-associated fibroblasts (CAFs)-they promote progression of different tumor types. We investigated the characteristics and functions of CAFs in diffuse-type gastric cancers (DGCs) by analyzing features of their genome and gene expression patterns. METHODS: We isolated CAFs and adjacent non-cancer fibroblasts (NFs) from 110 GC tissues from patients who underwent gastrectomy in Japan from 2008 through 2016...
April 5, 2017: Gastroenterology
https://www.readbyqxmd.com/read/28388924/enhancing-the-toolbox-to-study-il-17a-in-cattle-and-sheep
#12
Sean R Wattegedera, Yolanda Corripio-Miyar, Yvonne Pang, David Frew, Tom N McNeilly, Javier Palarea-Albaladejo, Colin J McInnes, Jayne C Hope, Elizabeth J Glass, Gary Entrican
The development of methods to detect cytokine expression by T cell subsets in ruminants is fundamental to strategic development of new livestock vaccines for prevention of infectious diseases. It has been possible to detect T cell expression of IFN-γ, IL-4 and IL-10 in ruminants for many years but methods to detect expression of IL-17A are relatively limited. To address this gap in capability we have cloned bovine and ovine IL-17A cDNAs and expressed biologically-active recombinant proteins in Chinese Hamster Ovary (CHO) cells...
April 8, 2017: Veterinary Research
https://www.readbyqxmd.com/read/28386188/optimizing-conditions-for-calcium-phosphate-mediated-transient-transfection
#13
Ling Guo, Liyang Wang, Ronghua Yang, Rui Feng, Zhongguang Li, Xin Zhou, Zhilong Dong, George Ghartey-Kwansah, MengMeng Xu, Miyuki Nishi, Qi Zhang, Williams Isaacs, Jianjie Ma, Xuehong Xu
BACKGROUND: Calcium phosphate mediated transfection has been used for delivering DNA into mammalian cells in excess of 30 years due to its most low cost for introducing recombinant DNA into culture cells. However, multiple factors affecting the transfect efficiency are commonly recognized meanwhile for years, the low transfection efficiency of this approach on higher differentiated and non-tumor cells such as CHO and C2C12 limits its application on research. RESULTS: In this paper, we systematically evaluated the possible factors affecting the transfection rate of this approach...
March 2017: Saudi Journal of Biological Sciences
https://www.readbyqxmd.com/read/28381637/aminoacyl-trna-synthetase-interacting-multifunctional-protein-1-activates-nk-cells-via-macrophages-in-vitro-and-in-vivo
#14
Myun Soo Kim, Ju Han Song, Edward P Cohen, Daeho Cho, Tae Sung Kim
Aminoacyl tRNA synthetase-interacting multifunctional protein 1 (AIMP1) has been reported to have antitumor effects in various tumor models. However, mechanisms by which AIMP1 ameliorates tumorigenesis are not well understood. As NK cells are a major cell type involved in antitumor activities and AIMP1 is known to activate professional APCs, we determined whether AIMP1 induced NK cell activation directly or via these APCs. AIMP1 induced the expression of surface activation markers on murine NK cells in total splenocytes, although AIMP1 did not directly induce these activation markers of NK cells...
April 5, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28378582/low-affinity-zinc-sensor-showing-fluorescence-responses-with-minimal-artifacts
#15
Xinhao Yan, Jin Ju Kim, Hye Sun Jeong, Yu Kyung Moon, Yoon Kyung Cho, Soyeon Ahn, Sang Beom Jun, Hakwon Kim, Youngmin You
The study of the zinc biology requires molecular probes with proper zinc affinity. We developed a low-affinity zinc probe (HBO-ACR) based on an azacrown ether (ACR) and an 2-(2-hydroxyphenyl)benzoxazole (HBO) fluorophore. This probe design imposed positive charge in the vicinity of a zinc coordination center, which enabled fluorescence turn-on responses to high levels of zinc without being affected by the pH and the presence of other transition-metal ions. Steady-state and transient photophysical investigations suggested that such a high tolerance benefits from orchestrated actions of proton-induced nonradiative and zinc-induced radiative control...
April 5, 2017: Inorganic Chemistry
https://www.readbyqxmd.com/read/28378146/use-of-a-highly-parallel-microfluidic-flow-cell-array-to-determine-therapeutic-drug-dose-response-curves
#16
Jesús A Arellano, Taylor A Howell, James Gammon, Sungpil Cho, Margit M Janát-Amsbury, Bruce Gale
A high-throughput, microfluidic flow cell array (MFCA) system has been modified to enable drug screening against small-volume cell-, and tissue cultures. The MFCA is composed of a 3D channel network that simultaneously flows fluids through forty-eight 830 μm by 500 μm flow cells, which physically divide and fluidically seal an existing culture into multiple compartments when docked onto the surface of a cell or tissue culture dish. The modified system provides temperature (37 °C) and CO2/pH level controls, while continuously flowing solutions (media or other liquid such as drug suspensions) over the cells/tissues...
June 2017: Biomedical Microdevices
https://www.readbyqxmd.com/read/28371311/low-glucose-concentrations-within-typical-industrial-operating-conditions-have-minimal-effect-on-the-transcriptome-of-recombinant-cho-cells
#17
Yogender Kumar Gowtham, Christopher A Saski, Sarah W Harcum
Typically, mammalian cell culture medium contains high glucose concentrations that are analogous to diabetic levels in humans, suggesting that mammalian cells are cultivated in excessive glucose. Using RNA-Seq, this study characterized the Chinese hamster ovary (CHO) cell transcriptome under two glucose concentrations to assess the genetic effects associated with metabolic pathways, in addition to other global responses. The initial extracellular glucose concentrations used represented high (30 mM) and low (10 mM) glucose conditions, where at the time the transcriptomes were compared, the glucose concentrations were approximately 24 and 4...
April 3, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28371246/long-term-liver-specific-functions-of-hepatocytes-in-electrospun-chitosan-nanofiber-scaffolds-coated-with-fibronectin
#18
Divya Rajendran, Ali Hussain, Derek Yip, Amit Parekh, Anil Shrirao, Cheul H Cho
In this study, a new 3-D liver model was developed using biomimetic nanofiber scaffolds and co-culture system consisting of hepatocytes and fibroblasts for the maintenance of long-term liver functions. The chitosan nanofiber scaffolds were fabricated by the electrospinning technique. To enhance cellular adhesion and spreading, the surfaces of the chitosan scaffolds were coated with fibronectin by adsorption and evaluated for various cell types. Cellular phenotype, protein expression, and liver-specific functions were extensively characterized by immunofluorescent and histochemical stainings, albumin ELISA and Cytochrome p450 detoxification assays, and SEM...
March 28, 2017: Journal of Biomedical Materials Research. Part A
https://www.readbyqxmd.com/read/28369783/utilization-of-sequence-variants-as-biomarkers-to-analyze-population-dynamics-in-cloned-cell-lines
#19
T Jennifer Lin, Kathryn M Beal, Heather S DeGruttola, Steven J Brennan, Lisa A Marzilli, Karin Anderson
The inherent nature of cloned CHO cell lines includes the presence of genetic and phenotypic drift that leads to heterogeneous populations. The genetic heterogeneity exhibited by these cells can be exploited to understand the population dynamics of cloned cell lines. Understanding the interplay between heterogeneity, cell culture conditions and population dynamics will allow for critical assessment of overarching cell line development methods and strategies in terms of population and monoclonality. Sequence variants (SVs) are protein isoforms of the gene-of-interest that contain unintended amino acid substitutions, extensions or truncations that may contribute to heterogeneity...
March 28, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28365978/establishment-of-immortalized-primary-human-foreskin-keratinocytes-and-their-application-to-toxicity-assessment-and-three-dimensional-skin-culture-construction
#20
Moonju Choi, Minkyung Park, Suhyon Lee, Jeong Woo Lee, Min Chul Cho, Minsoo Noh, Choongho Lee
In spite of frequent usage of primary human foreskin keratinocytes (HFKs) in the study of skin biology, senescence-induced blockage of in vitro proliferation has been a big hurdle for their effective utilization. In order to overcome this passage limitation, we first isolated ten HFK lines from circumcision patients and successfully immortalized four of them via a retroviral transduction of high-risk human papillomavirus (HPV) E6 and E7 oncogenes. We confirmed expression of a keratinocyte marker protein, keratin 14 and two viral oncoproteins in these immortalized HFKs...
May 1, 2017: Biomolecules & Therapeutics
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