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https://www.readbyqxmd.com/read/28212566/genetic-characterization-of-polish-ccrcc-patients-somatic-mutation-analysis-of-pbrm1-bap1-and-kdmc5-genomic-snp-array-analysis-in-tumor-biopsy-and-preliminary-results-of-chromosome-aberrations-analysis-in-plasma-cell-free-dna
#1
Katarzyna Kluzek, Malgorzata I Srebniak, Weronika Majer, Agnieszka Ida, Tomasz Milecki, Kinga Huminska, Robert M van der Helm, Adrian Silesian, Tomasz M Wrzesinski, Jacek Wojciechowicz, Berna H Beverloo, Zbigniew Kwias, Hans A R Bluyssen, Joanna Wesoly
BACKGROUND: Mutation analysis and cytogenetic testing in clear cell renal cell carcinoma (ccRCC) is not yet implemented in a routine diagnostics of ccRCC. MATERIAL AND METHODS: We characterized the chromosomal alterations in 83 ccRCC tumors from Polish patients using whole genome SNP genotyping assay. Moreover, the utility of next generation sequencing of cell free DNA (cfDNA) in patients plasma as a potential tool for non-invasive cytogenetic analysis was tested...
February 15, 2017: Oncotarget
https://www.readbyqxmd.com/read/28212557/using-high-sensitivity-sequencing-for-the-detection-of-mutations-in-btk-and-plc%C3%AE-2-genes-in-cellular-and-cell-free-dna-and-correlation-with-progression-in-patients-treated-with-btk-inhibitors
#2
Adam Albitar, Wanlong Ma, Ivan DeDios, Jeffrey Estella, Inhye Ahn, Mohammed Farooqui, Adrian Wiestner, Maher Albitar
Patients with chronic lymphocytic leukemia (CLL) that develop resistance to Bruton tyrosine kinase (BTK) inhibitors are typically positive for mutations in BTK or phospholipase c gamma 2 (PLCγ2). We developed a high sensitivity (HS) assay utilizing wild-type blocking polymerase chain reaction achieved via bridged and locked nucleic acids. We used this high sensitivity assay in combination with Sanger sequencing and next generation sequencing (NGS) and tested cellular DNA and cell-free DNA (cfDNA) from patients with CLL treated with the BTK inhibitor, ibrutinib...
February 13, 2017: Oncotarget
https://www.readbyqxmd.com/read/28210900/false-responses-of-renilla-luciferase-reporter-control-to-nuclear-receptor-tr4
#3
Dongyun Zhang, Sam S Atlasi, Krishna K Patel, Zihao Zhuang, Anthony P Heaney
Renilla luciferase reporter is a widely used internal control in dual luciferase reporter assay system, where its transcription is driven by a constitutively active promoter. However, the authenticity of the Renilla luciferase response in some experimental settings has recently been questioned. Testicular receptor 4 (TR4, also known as NR2C2) belongs to the subfamily 2 of nuclear receptors. TR4 binds to a direct repeat regulatory element in the promoter of a variety of target genes and plays a key role in tumorigenesis, lipoprotein regulation, and central nervous system development...
February 16, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28209555/detection-of-soluble-erbb2-in-breast-cancer-cell-lysates-using-a-combined-label-free-fluorescence-platform-based-on-bloch-surface-waves
#4
Alberto Sinibaldi, Camilla Sampaoli, Norbert Danz, Peter Munzert, Leonardo Sibilio, Frank Sonntag, Agostino Occhicone, Elisabetta Falvo, Elisa Tremante, Patrizio Giacomini, Francesco Michelotti
We report on the use of one-dimensional photonic crystals to detect clinically relevant concentrations of ERBB2/neu/Her2 in cell lysates. ERBB2 is a pivotal breast cancer biomarker and targetable oncogenic driver associated with aggressive breast cancer subtypes. To quantitate soluble ERBB2, we developed an optical platform that combines label-free and fluorescence detection modes. Such platform makes use of a sandwich assay in which the one-dimensional photonic crystals sustaining Bloch surface waves are tailored with a monoclonal antibody for highly specific biological recognition (BSW biochip)...
February 9, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28208613/structure-antioxidative-and-anti-inflammatory-activity-relationships-of-purpurin-and-related-anthraquinones-in-chemical-and-cell-assays
#5
Woo Nam, Sung Phil Kim, Seok Hyun Nam, Mendel Friedman
Anthraquinone (9,10-anthraquinone) and several hydroxy derivatives, including purpurin (1,2,4-trihydroxyanthraquinone), anthrarufin (1,5-dihydroxyanthraquinone), and chrysazin (1,8-dihydroxyanthraquinone), were evaluated for antioxidative and anti-inflammatory activities in chemical assays and mammalian cells (murine macrophage RAW 264.7 cells). Several tests were used to assess their activities: 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical; ABTS radical cation; hydrogen peroxide scavenging; reduction of potassium ferricyanide; chelation of ferrous ions; inhibition of lipid peroxidation; inhibition of nitric oxide generation; scavenging of the intracellular hydroxyl radical; expression of NLRP3 polypeptide for inflammasome assembly; and quantitation of proinflammatory cytokine interleukin 1β (IL-1β) for inflammasome activation...
February 10, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/28208291/growth-based-bacterial-viability-assay-for-interference-free-and-high-throughput-toxicity-screening-of-nanomaterials
#6
Tian A Qiu, Thu Ha Thi Nguyen, Natalie V Hudson-Smith, Peter L Clement, Dona-Carla Forester, Hilena Frew, Mimi N Hang, Catherine J Murphy, Robert J Hamers, Z Vivian Feng, Christy L Haynes
Current high-throughput approaches evaluating toxicity of chemical agents toward bacteria typically rely on optical assays, such as luminescence and absorbance, to probe the viability of the bacteria. However, when applied to toxicity induced by nanomaterials, scattering and absorbance from the nanomaterials act as interferences that complicate quantitative analysis. Herein, we describe a bacterial viability assay that is free of optical interference from nanomaterials and can be performed in a high-throughput format on 96-well plates...
February 7, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28208281/enzyme-activated-g-quadruplex-synthesis-for-in-situ-label-free-detection-and-bioimaging-of-cell-apoptosis
#7
Zhuoliang Liu, Xingyu Luo, Zhu Li, Yan Huang, Zhou Nie, Hong-Hui Wang, Shouzhuo Yao
Fluorogenic probes targeting G-quadruplex structures have emerged as the promising toolkit for functional research of G-quadruplex and biosensor development. However, their biosensing applications are still largely limited in in-tube detection. Herein, we proposed a fluorescent bioimaging method based on enzyme-generated G-quadruplexes for detecting apoptotic cells at the cell and tissue level, namely, terminal deoxynucleotidyl transferase (TdT)-activated de novo G-quadruplex synthesis (TAGS) assay. The detection target is genomic DNA fragmentation, a biochemical hallmark of apoptosis...
February 7, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28207992/inhibitory-activities-of-n-butanol-fraction-from-butea-monosperma-lam-taub-bark-against-free-radicals-genotoxins-and-cancer-cells
#8
Varinder Kaur, Manish Kumar, Paramjeet Kaur, Sandeep Kaur, Satwinderjeet Kaur
The present study was undertaken to investigate antioxidant, antigenotoxic and antiproliferative activity of n-butanol fraction (Bmbu) from bark of medicinal plant Butea monosperma. Antioxidant potency of Bmbu was examined by various in vitro assays. It was also investigated for antigenotoxic activity using E. coli. PQ37 employing SOS chromotest. Further, cytotoxic and apoptosis inducing activity of Bmbu was evaluated in MCF-7 breast cancer cells. Bmbu showed potent free radical scavenging ability in ABTS assay (IC50 56...
February 16, 2017: Chemistry & Biodiversity
https://www.readbyqxmd.com/read/28206954/rassf1a-promoter-methylation-in-high-grade-serous-ovarian-cancer-a-direct-comparison-study-in-primary-tumors-adjacent-morphologically-tumor-cell-free-tissues-and-paired-circulating-tumor-dna
#9
Lydia Giannopoulou, Issam Chebouti, Kitty Pavlakis, Sabine Kasimir-Bauer, Evi S Lianidou
The RASSF1A promoter is frequently methylated in high-grade serous ovarian cancer (HGSC). We examined RASSF1A promoter methylation in primary tumors, adjacent morphologically tumor cell-free tissues and corresponding circulating tumor DNA (ctDNA) samples of patients with HGSC, using a real-time methylation specific PCR (real-time MSP) and a methylation-sensitive high-resolution melting analysis (MS-HRMA) assay for the detection and semi-quantitative estimation of methylation, respectively. Two groups of primary HGSC tumor FFPE samples were recruited (Group A n=67 and Group B n=61), along with matched adjacent morphologically tumor cell-free tissues (n=58) and corresponding plasma samples (n=59) for group B...
February 10, 2017: Oncotarget
https://www.readbyqxmd.com/read/28205434/recent-advances-on-magnetic-relaxation-switching-assay-based-nanosensors
#10
Yang Zhang, Hong Yang, Zhiguo Zhou, Kai Huang, Shi-Ping Yang, Gang Han
Magnetic relaxation switching assay (MRSw)-based nanosensors respond to the changes of transverse relaxation time (T2) of water molecules resulted from the analyte-induced aggregation / disaggregation of magnetic nanoparticles (MNPs). This strategy has been widely applied to the detections of various substrates from heavy metal ions to organic pollutants, proteins, nucleic acids, bacteria/viruses, and specific cells. Compared with other nanosensors, MRSw-based nanosensors not only are free from the background interferences, signal bleaching, and quenching but also overcome light scattering from samples without pre-treatments...
February 16, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28205180/methods-for-monitoring-abca1-dependent-sterol-release
#11
Yoshio Yamauchi, Shinji Yokoyama, Ta-Yuan Chang
Releasing sterols to the extracellular milieu is an important part of sterol homeostasis in cells and in the body. ATP-binding cassette transporter A1 (ABCA1) plays an essential role in cellular phospholipid and sterol release to lipid-free or lipid-poor apolipoprotein A-I (apoA-I), the major apolipoprotein in high-density lipoprotein (HDL), and constitutes the first step in the formation of nascent HDL. Loss-of-function mutations in the ABCA1 gene lead to a rare disease known as Tangier disease that causes severe deficiency in plasma HDL level...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28205023/low-serum-culture-with-novel-medium-promotes-maxillary-mandibular-bone-marrow-stromal-cell-proliferation-and-osteogenic-differentiation-ability
#12
Fumio Suehiro, Masakazu Ishii, Izumi Asahina, Hiroshi Murata, Masahiro Nishimura
OBJECTIVES: The purpose of this study was to evaluate the effect of low-serum STK2 medium on the isolation and osteogenic differentiation of human maxillary/mandibular bone marrow stromal cells (MBMSCs). MATERIALS AND METHODS: Human MBMSCs were obtained from patients undergoing dental implant treatment. These cells were cultured in serum-free medium or STK2 medium containing 1  % fetal bovine serum (low-serum) or α-MEM containing 10  % fetal bovine serum (control)...
February 16, 2017: Clinical Oral Investigations
https://www.readbyqxmd.com/read/28202289/rolipram-potentiates-bevacizumab-induced-cell-death-in-human-glioblastoma-stem-like-cells
#13
Sara Ramezani, Nasim Vousooghi, Fatemeh Ramezani Kapourchali, Mahmoudreza Hadjighasem, Parisa Hayat, Naser Amini, Mohammad Taghi Joghataei
AIMS: Glioblastoma cancer stem-like cells (GCSCs) promote themselves proliferation by secreting the vascular endothelial growth factor A (VEGFA) in an autocrine manner, positively regulated by phosphodiesterase IV (PDE4). In the current study, we investigated the putative cytotoxic effect of bevacizumab, a VEGFA blocker, alone and in combination with a specific inhibitor of PDE4 called rolipram on GCSCs isolated from human surgical tumor specimen with a focus on PI3K/AKT pathway. MAIN METHODS: CD133+/CD15+ GCSCs were characterized by flow cytometry and expanded in a serum-free primary culture system...
February 12, 2017: Life Sciences
https://www.readbyqxmd.com/read/28199863/analysis-of-chemopredictive-assay-for-targeting-cancer-stem-cells-in-glioblastoma-patients
#14
Candace M Howard, Jagan Valluri, Anthony Alberico, Terrence Julien, Rida Mazagri, Robert Marsh, Hoyt Alastair, Antonio Cortese, Michael Griswold, Wanmei Wang, Krista Denning, Linda Brown, Pier Paolo Claudio
INTRODUCTION: The prognosis of glioblastoma (GBM) treated with standard-of-care maximal surgical resection and concurrent adjuvant temozolomide (TMZ)/radiotherapy remains very poor (less than 15 months). GBMs have been found to contain a small population of cancer stem cells (CSCs) that contribute to tumor propagation, maintenance, and treatment resistance. The highly invasive nature of high-grade gliomas and their inherent resistance to therapy lead to very high rates of recurrence. For these reasons, not all patients with similar diagnoses respond to the same chemotherapy, schedule, or dose...
February 12, 2017: Translational Oncology
https://www.readbyqxmd.com/read/28197649/dose-response-relationship-of-temozolomide-determined-by-the-pig-a-comet-and-micronucleus-assay
#15
M Guérard, G Johnson, S Dertinger, G Duran-Pacheco, J Funk, A Zeller
Temozolomide (TMZ), a monofunctional alkylating agent, was selected as a model compound to determine its quantitative genotoxic dose-response relationship in different tissues (blood, liver, and jejunum) and endpoints [Pig-a-, comet-, and micronucleus assay (MNT)] in male rats. TMZ was administered p.o. over 5 consecutive days (day 1-5), followed by a treatment-free period of 50 days (day 6-56) and a final administration prior to necropsy (day 57-59). TMZ showed a dose-dependent increase in DNA damage in all interrogated endpoints...
February 15, 2017: Archives of Toxicology
https://www.readbyqxmd.com/read/28196921/noninvasive-prenatal-screening-of-fetal-aneuploidy-without-massively-parallel-sequencing
#16
Chenming Xu, Ting Wang, Chao Liu, Hong Li, Xiaoyan Chen, Huanhuan Zhu, Songchang Chen, Qiuhong Xin, Jing Tao, Liming Huang, Zhengwen Jiang
BACKGROUND: Noninvasive prenatal screening (NIPS) using plasma cell-free DNA has gained tremendous popularity in the clinical assessment of fetal aneuploidy. Most, if not all, of these tests rely on complex and expensive massively parallel sequencing (MPS) techniques, hindering the use of NIPS as a common screening procedure. METHODS: We have developed and optimized an MPS-independent noninvasive genetic test that can rapidly detect fetal aneuploidy at considerably lower costs...
February 14, 2017: Clinical Chemistry
https://www.readbyqxmd.com/read/28196836/label-free-dynamic-mass-redistribution-reveals-low-density-pro-survival-%C3%AE-1b-adrenergic-receptors-in-human-sw480-colon-carcinoma-cells
#17
Dorathy-Ann Harris, Ji-Min Park, Kyung Soon-Lee, Cong Xu, Nephi Stella, Chris Hague
Small molecules that target the adrenergic family of G protein-coupled receptors (GPCRs) show promising therapeutic efficacy for the treatment of various cancers. Herein, we report that human colon cancer cell line SW480 expresses low-density functional α1B-adrenergic receptors (ARs) as revealed by label-free dynamic mass redistribution (DMR) signaling technology and confirmed by quantitative reverse-transcriptase polymerase chain reaction analysis. Remarkably, while endogenous α1B-ARs are not detectable via either [3H]-prazosin binding analysis or phosphoinositol hydrolysis assays, their activation leads to robust DMR and enhanced cell viability...
February 14, 2017: Journal of Pharmacology and Experimental Therapeutics
https://www.readbyqxmd.com/read/28193039/molecularly-imprinted-polymer-based-plasmonic-immunosandwich-assay-for-fast-and-ultrasensitive-determination-of-trace-glycoproteins-in-complex-samples
#18
Xueying Tu, Pir Muhammad, Jia Liu, Yanyan Ma, Shuangshou Wang, Danyang Yin, Zhen Liu
Glycoproteins play significant roles in many biological processes. Assays of glycoproteins have significant biological importance and clinical values, for which immunoassay has been the workhorse tool. However, immunoassay suffers from some disadvantages, such as poor availability of high-specificity antibodies and limited stability of biological reagents. Herein, we present an antibody-free and enzyme-free approach, called molecularly imprinted polymer (MIP)-based plasmonic immunosandwich assay (PISA), for fast and ultrasensitive detection of trace glycoproteins in complex samples...
December 20, 2016: Analytical Chemistry
https://www.readbyqxmd.com/read/28193027/fluorescence-imaging-of-intracellular-telomerase-activity-using-enzyme-free-signal-amplification
#19
Min Hong, Lidan Xu, Qingwang Xue, Lu Li, Bo Tang
A novel enzyme-free signal amplification-based assay for highly sensitive in situ fluorescence imaging and detection of intracellular telomerase activity was developed by using a gold nanoflare probe-triggered mimic-hybridization chain reaction (mimic-HCR) coupled with a graphene oxide (GO) surface-anchored fluorescence signal readout pathway. The nanoflare probe consists of gold nanoparticles (AuNPs) functionalized with a dense shell of nucleic acid sequences by Au-S bond formation. The nucleic acid sequence is composed of three segments: a long thiol-labeled sequence (HS-DNA) and two short sequences (a telomerase primer sequence, "Primer-DNA", and an FAM-terminated reporter sequence, "Flare-DNA"), both of which are complementary to HS-DNA...
December 20, 2016: Analytical Chemistry
https://www.readbyqxmd.com/read/28192955/microfluidic-migration-and-wound-healing-assay-based-on-mechanically-induced-injuries-of-defined-and-highly-reproducible-areas
#20
Drago Sticker, Sarah Lechner, Christian Jungreuthmayer, Jürgen Zanghellini, Peter Ertl
All cell migration and wound healing assays are based on the inherent ability of adherent cells to move into adjacent cell-free areas, thus providing information on cell culture viability, cellular mechanisms and multicellular movements. Despite their widespread use for toxicological screening, biomedical research and pharmaceutical studies, to date no satisfactory technological solutions are available for the automated, miniaturized and integrated induction of defined wound areas. To bridge this technological gap, we have developed a lab-on-a-chip capable of mechanically inducing circular cell-free areas within confluent cell layers...
January 31, 2017: Analytical Chemistry
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