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https://www.readbyqxmd.com/read/28735045/multiplexed-capillary-microfluidic-immunoassay-with-smartphone-data-acquisition-for-parallel-mycotoxin-detection
#1
Jessica M D Machado, Ruben R G Soares, Virginia Chu, João P Conde
The field of microfluidics holds great promise for the development of simple and portable lab-on-a-chip systems. The use of capillarity as a means of fluidic manipulation in lab-on-a-chip systems can potentially reduce the complexity of the instrumentation and allow the development of user-friendly devices for point-of-need analyses. In this work, a PDMS microchannel-based, colorimetric, autonomous capillary chip provides a multiplexed and semi-quantitative immunodetection assay. Results are acquired using a standard smartphone camera and analyzed with a simple gray scale quantification procedure...
July 15, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28734694/regenerative-biosensor-for-use-with-biotinylated-bait-molecules
#2
Claudia Knoglinger, Andreas Zich, Lukas Traxler, Kristýna Poslední, Gloria Friedl, Bianca Ruttmann, Anika Schorpp, Katharina Müller, Mirjam Zimmermann, Hermann J Gruber
Label-free biosensors are ideally suited for the quantitative analysis of specific interactions among biomolecules or of biomolecules with drugs, as well as for quantitation of diagnostic markers in biofluids. In contrast to the label-dependent methods, a new assay for a particular prey molecule can be set up within few minutes by immobilizing the corresponding bait molecule on the sensor surface, using one of the common immobilization procedures. Unfortunately, the extensive application of label-free biosensors is still hampered by the fact that the immobilization of the bait molecule is usually irreversible; for that reason, a new chip (which is expensive) is required for every successful or futile attempt...
July 13, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28734156/a-novel-hypoxia-response-element-regulates-oxygen-related-repression-of-tissue-factor-pathway-inhibitor-in-the-breast-cancer-cell-line-mcf-7
#3
Xue Yan Cui, Grethe Skretting, Mari Tinholt, Benedicte Stavik, Anders Erik Astrup Dahm, Kristine Kleivi Sahlberg, Sandip Kanse, Nina Iversen, Per Morten Sandset
BACKGROUND: Hypoxia is one of the most pervasive physiological stresses in solid tumors. We have previously demonstrated that tissue factor (TF) pathway inhibitor (TFPI) expression was transcriptionally repressed by the activation of hypoxia inducible factor (HIF)-1α under hypoxic conditions. However, the role of HIF-2α, also known as endothelial PAS domain-containing protein 1 (EPAS1), on TFPI expression remains unclear. AIM: To explore the role of HIF-2α/EPAS1 in the regulation of TFPI expression under hypoxia in breast cancer cells...
July 14, 2017: Thrombosis Research
https://www.readbyqxmd.com/read/28733419/myc2-orchestrates-a-hierarchical-transcriptional-cascade-that-regulates-jasmonate-mediated-plant-immunity-in-tomato
#4
Minmin Du, Jiuhai Zhao, David T W Tzeng, Yuanyuan Liu, Lei Deng, Tianxia Yang, Qingzhe Zhai, Fangming Wu, Zhuo Huang, Ming Zhou, Qiaomei Wang, Qian Chen, Silin Zhong, Chang-Bao Li, Chuanyou Li
The hormone jasmonate (JA), which functions in plant immunity, regulates resistance to pathogen infection and insect attack through triggering genome-wide transcriptional reprogramming in plants. We show that the basic helix-loop-helix transcription factor (TF) MYC2 in tomato (Solanum lycopersicum) acts downstream of the JA receptor to orchestrate JA-mediated activation of both the wounding and pathogen responses. Using chromatin immunoprecipitation sequencing (ChIP-seq) coupled with RNA sequencing (RNA-seq) assays, we identified 655 MYC2-targeted JA-responsive genes...
July 21, 2017: Plant Cell
https://www.readbyqxmd.com/read/28731191/carboxy-terminus-hsc70-interacting-protein-exerts-a-tumor-inhibition-function-in-head-and-neck-cancer
#5
Meng Xiao, Ming Yan, Jianjun Zhang, Qin Xu, Wantao Chen
Several independent studies have reported the roles of the E3 ubiquitin ligase, carboxy-terminus Hsc70 interacting protein (CHIP) in various types of cancers. However, the biological effects of CHIP vary in regards to different cancers, and the role of CHIP in head and neck cancers (HNCs) remains unknown. In the present study, CHIP overexpression plasmids and CHIP knockdown lentivirus were constructed to affect the expression levels of CHIP protein and biological behaviors in HNC cell lines bilaterally. The biological behaviors regulated by CHIP in HNCs were investigated both in vivo and in vitro with a series of assays and analyses...
July 17, 2017: Oncology Reports
https://www.readbyqxmd.com/read/28726915/oleuropein-represses-the-radiation-resistance-of-ovarian-cancer-by-inhibiting-hypoxia-and-microrna-299-targetted-heparanase-expression
#6
Yubai Xing, Dawei Cui, Shun Wang, Pingping Wang, Xiaojing Xing, Hui Li
Radiotherapy in ovarian cancer frequently invokes resistance; this severely compromises its therapeutic effect and results in poor clinical prognosis. How to overcome the acquired resistance and re-sensitize tumors to radiation is the central question in this clinical setting. Cancer cell survival was evaluated using a clonogenic assay. The microRNA expression profile was analyzed using a microarray. Transcript expression was determined using real time PCR. The expression of protein was determined by immunoblotting...
July 20, 2017: Food & Function
https://www.readbyqxmd.com/read/28726847/mapping-genome-wide-transcription-factor-binding-sites-using-dap-seq
#7
Anna Bartlett, Ronan C O'Malley, Shao-Shan Carol Huang, Mary Galli, Joseph R Nery, Andrea Gallavotti, Joseph R Ecker
To enable low-cost, high-throughput generation of cistrome and epicistrome maps for any organism, we developed DNA affinity purification sequencing (DAP-seq), a transcription factor (TF)-binding site (TFBS) discovery assay that couples affinity-purified TFs with next-generation sequencing of a genomic DNA library. The method is fast, inexpensive, and more easily scaled than chromatin immunoprecipitation sequencing (ChIP-seq). DNA libraries are constructed using native genomic DNA from any source of interest, preserving cell- and tissue-specific chemical modifications that are known to affect TF binding (such as DNA methylation) and providing increased specificity as compared with in silico predictions based on motifs from methods such as protein-binding microarrays (PBMs) and systematic evolution of ligands by exponential enrichment (SELEX)...
August 2017: Nature Protocols
https://www.readbyqxmd.com/read/28726375/nanoscale-hydrodynamic-film-for-diffusive-mass-transport-control-in-compartmentalized-microfluidic-chambers
#8
Minseok Kim, Ji Won Lim, Sung Kuk Lee, Taesung Kim
A compartmentalized microfluidic chamber array that not only offers separate cell culture environments but also independent control of the diffusion of small molecules provides an extremely useful platform for cell cultivations and versatile cellular assays. However, it is challenging to incorporate both cell compartmentalization and active diffusion control in real-time and precise manners. Here, we present a novel nanoscale hydrodynamic film (NHF) that is formed between a solid substrate and a polydimethylsiloxane (PDMS) surface...
July 20, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28723624/lncrna-ancr-down-regulation-promotes-tgf-%C3%AE-induced-emt-and-metastasis-in-breast-cancer
#9
Zhongwei Li, Meichen Dong, Dongmei Fan, Pingfu Hou, Hongyuan Li, Lingxia Liu, Cong Lin, Jiwei Liu, Liangping Su, Lan Wu, Xiaoxue Li, Baiqu Huang, Jun Lu, Yu Zhang
Epithelial to mesenchymal transition (EMT) is a progression of cellular plasticity critical for development, differentiation, cancer cells migration and tumor metastasis. As a well-studied factor, TGF-β participates in EMT and involves in physiological and pathological functions of tumor progression. Accumulating evidence indicates that long noncoding RNAs(lncRNAs) play crucial roles in EMT and tumor metastasis. Here, we find that lncRNA ANCR participates in TGF-β1-induced EMT. By our ChIP and Real-time PCR assays, we reveal that TGF-β1 down-regulates ANCR expression by increasing HDAC3 enrichment at ANCR promoter region, which decreases both H3 and H4 acetylation of ANCR promoter...
June 27, 2017: Oncotarget
https://www.readbyqxmd.com/read/28723184/steric-hindrance-assay-for-secreted-factors-in-stem-cell-culture
#10
Wendi Zhou, Sahar S Mahshid, Weijia Wang, Alexis Vallée-Bélisle, Peter W Zandstra, Edward H Sargent, Shana O Kelley
The ex vivo expansion of hematopoietic stem cells is significantly inhibited by secreted proteins that induce negative feedback loops. The ability to effectively monitor these factors is critical for their real-time regulation and control and, by extension, enhancing stem cell expansion. Here, we describe a novel monitoring strategy for the detection of soluble signaling factors in stem cell cultures using a DNA-based sensing mechanism on a chip-based nanostructured microelectrode platform. We combine DNA hybridization engineering with antibody-capturing chemistry in an amplified steric hindrance hybridization assay...
April 28, 2017: ACS Sensors
https://www.readbyqxmd.com/read/28719742/integrated-smartphone-app-chip-system-for-on-site-ppb-level-colorimetric-quantitation-of-aflatoxins
#11
Xiaochun Li, Fan Yang, Jessica X H Wong, Hua-Zhong Yu
We demonstrate herein an integrated, smartphone-app-chip (SPAC) system for on-site quantitation of food toxins, as demonstrated with aflatoxin B1 (AFB1), at parts-per-billion (ppb) level in food products. The detection is based on an indirect competitive immunoassay fabricated on a transparent plastic chip with the assistance of a microfluidic channel plate. A 3D-printed optical accessory attached to a smartphone is adapted to align the assay chip and to provide uniform illumination for imaging, with which high-quality images of the assay chip are captured by the smartphone camera and directly processed using a custom-developed Android app...
July 18, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28716136/up-regulation-of-cdk16-by-multiple-mechanisms-in-hepatocellular-carcinoma-promotes-tumor-progression
#12
Yitao Wang, Xian Qin, Tao Guo, Pengpeng Liu, Ping Wu, Zhisu Liu
BACKGROUND: Hepatocellular carcinoma (HCC) remains difficult to cure due to lack of effective treatment and the molecular mechanisms are complex and not completely understood. In this study, We investigated the role of CDK16 in tumor progression of HCC. METHODS: We interrogated the expression level of CDK16 by polymerase chain reaction and immunohistochemistry(IHC) and studied its clinical significance. The functional role of CDK16 on HCC was studied via gain and loss of function in vitro and in vivo...
July 17, 2017: Journal of Experimental & Clinical Cancer Research: CR
https://www.readbyqxmd.com/read/28715808/retinoic-acid-facilitates-toll-like-receptor-4-expression-to-improve-intestinal-barrier-function-through-retinoic-acid-receptor-beta
#13
Yingying Li, Yuan Gao, Ting Cui, Ting Yang, Lan Liu, Tingyu Li, Jie Chen
BACKGROUND/AIMS: Vitamin A (VA) protects the intestinal epithelial barrier by improving cell migration and proliferation. Our previous studies demonstrated that VA deficiency (VAD) during pregnancy suppresses the systemic and mucosal immune responses in the intestines of offspring and that VA supplementation (VAS) during early life can increase immune cell counts. However, little is known about the mechanisms by which VA regulates intestinal epithelial barrier function. METHODS: Caco-2 cells were treated with all-trans retinoic acid (ATRA) for 24 hours to determine the optimum ATRA concentration to which the cells in question respond...
July 17, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28715394/a-filtration-based-method-of-preparing-high-quality-nuclei-from-cross-linked-skeletal-muscle-for-chromatin-immunoprecipitation
#14
Kazunari Nohara, Zheng Chen, Seung-Hee Yoo
Chromatin immunoprecipitation (ChIP) is a powerful method to determine protein binding to chromatin DNA. Fiber-rich skeletal muscle, however, has been a challenge for ChIP due to technical difficulty in isolation of high-quality nuclei with minimal contamination of myofibrils. Previous protocols have attempted to purify nuclei before cross-linking, which incurs the risk of altered DNA-protein interaction during the prolonged nuclei preparation process. In the current protocol, we first cross-linked the skeletal muscle tissue collected from mice, and the tissues were minced and sonicated...
July 6, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28715365/analysis-of-the-c-kit-ligand-promoter-using-chromatin-immunoprecipitation
#15
Pingyu Zhang, Andres Rojas, Boris Blechacz
Multiple cellular processes, including DNA replication and repair, DNA recombination, and gene expression, require interactions between proteins and DNA. Therefore, DNA-protein interactions regulate multiple physiological, pathophysiological, and biological functions, such as cell differentiation, cell proliferation, cell cycle control, chromosome stability, epigenetic gene regulation, and cell transformation. In eukaryotic cells, the DNA interacts with histone and nonhistone proteins and is condensed into chromatin...
June 27, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28714515/elncrna1-a-long-non-coding-rna-that-is-transcriptionally-induced-by-oestrogen-promotes-epithelial-ovarian-cancer-cell-proliferation
#16
Jun-Jun Qiu, Xiao-Dan Zhang, Xiao-Yan Tang, Ting-Ting Zheng, Ying Zhang, Ke-Qin Hua
Previously, the novel oestrogen (E2)-upregulated lncRNA TC0101441, was identified by us, via microarray analysis. However, the detailed mechanism by which E2 upregulates TC0101441 and the role of TC0101441 in epithelial ovarian cancer (EOC) progression have not been elucidated. In the present study, we further analysed TC0101441, which we designated oestrogen-induced long non-coding RNA-1 (ElncRNA1). We showed that E2 transcriptionally upregulates ElncRNA1 through the oestrogen receptor α (ERα)-oestrogen response element (ERE) pathway using RNA stability assays, bioinformatics-based searches for ERE binding sites, chromatin immunoprecipitation (ChIP) assays and dual luciferase reporter assays...
August 2017: International Journal of Oncology
https://www.readbyqxmd.com/read/28704506/microfluidic-assay-of-circulating-endothelial-cells-in-coronary-artery-disease-patients-with-angina-pectoris
#17
Shuiyu Chen, Yukun Sun, Kuang Hong Neoh, Anqi Chen, Weiju Li, Xiaorui Yang, Ray P S Han
BACKGROUND: Circulating endothelial cells (CECs) are widely reported as a promising biomarker of endothelial damage/dysfunction in coronary artery disease (CAD). The two popular methods of CEC quantification include the use of immunomagnetic beads separation (IB) and flow cytometry analysis (FC); however, they suffer from two main shortcomings that affect their diagnostic and prognostic responses: non-specific bindings of magnetic beads to non-target cells and a high degree of variability in rare cell identification, respectively...
2017: PloS One
https://www.readbyqxmd.com/read/28702612/rapid-label-free-cd4-testing-using-a-smartphone-compatible-device
#18
Manoj Kumar Kanakasabapathy, Hardik J Pandya, Mohamed Shehata Draz, Manjyot Kaur Chug, Magesh Sadasivam, Shreya Kumar, Behzad Etemad, Vinish Yogesh, Mohammadali Safavieh, Waseem Asghar, Jonathan Z Li, Athe M Tsibris, Daniel R Kuritzkes, Hadi Shafiee
The most recent guidelines have called for a significant shift towards viral load testing for HIV/AIDS management in developing countries; however point-of-care (POC) CD4 testing still remains an important component of disease staging in multiple developing countries. Advancements in micro/nanotechnologies and consumer electronics have paved the way for mobile healthcare technologies and the development of POC smartphone-based diagnostic assays for disease detection and treatment monitoring. Here, we report a simple, rapid (30 minutes) smartphone-based microfluidic chip for automated CD4 testing using a small volume (30 μL) of whole blood...
July 12, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28702328/ask1-map3k5-is-transcriptionally-upregulated-by-e2f1-in-adipose-tissue-in-obesity-molecularly-defining-a-human-dys-metabolic-obese-phenotype
#19
Yulia Haim, Matthias Blüher, Daniel Konrad, Nir Goldstein, Nora Klöting, Ilana Harman-Boehm, Boris Kirshtein, Doron Ginsberg, Tanya Tarnovscki, Yftach Gepner, Iris Shai, Assaf Rudich
OBJECTIVE: Obesity variably disrupts human health, but molecular-based patients' health-risk stratification is limited. Adipose tissue (AT) stresses may link obesity with metabolic dysfunction, but how they signal in humans remains poorly-characterized. We hypothesized that a transcriptional AT stress-signaling cascade involving E2F1 and ASK1 (MAP3K5) molecularly defines high-risk obese subtype. METHODS: ASK1 expression in human AT biopsies was determined by real-time PCR analysis, and chromatin immunoprecipitation (ChIP) adopted to AT explants was used to evaluate the binding of E2F1 to the ASK1 promoter...
July 2017: Molecular Metabolism
https://www.readbyqxmd.com/read/28701588/igfbp2-plays-an-important-role-in-heat-shock-protein-27-mediated-cancer-progression-and-metastasis
#20
Chin-Sheng Hung, Chien-Yu Huang, Chia-Hwa Lee, Wei-Yu Chen, Ming-Te Huang, Po-Li Wei, Yu-Jia Chang
Heat shock protein 27 (Hsp27) is a key chaperone that interacts with over 200 client proteins. The expression of Hsp27 might be correlated with poor outcome in many types of cancer. Previous study indicated that Hsp27 might be an important biomarker in hepatocellular carcinoma (HCC). However, the detailed mechanism is less well understood. The shRNA-mediated silencing of Hsp27 decreased the proliferation, migration and invasion of HCC cells. In a xenograft model, the silencing of Hsp27 reduced tumor progression...
July 5, 2017: Oncotarget
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