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https://www.readbyqxmd.com/read/28936657/whole-mount-rna-fish-on-ovules-and-developing-seeds
#1
Andrea Bleckmann, Thomas Dresselhaus
A key element to understand developmental and reproductive processes like germline development, double fertilization, and embryogenesis is the study of cell-specific gene expression patterns which is best analyzed by RNA in situ hybridization. Different visualization techniques have been established to mark either the region of mRNA production (using the classical chromogenic detection system) or the specific localization of mRNAs (using fluorescent labeled probes). In this chapter, we describe and compare whole mount RNA in situ hybridization techniques on ovules and young developing seeds from Arabidopsis thaliana using three different detection systems...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28934307/amplification-of-the-egfr-gene-can-be-maintained-and-modulated-by-variation-of-egf-concentrations-in-in-vitro-models-of-glioblastoma-multiforme
#2
Doreen William, Poroshista Mokri, Nora Lamp, Michael Linnebacher, Carl Friedrich Classen, Andreas Erbersdobler, Björn Schneider
Glioblastoma multiforme (GBM) is the most common and lethal brain tumor in adults. It is known that amplification of the epidermal growth factor receptor gene (EGFR) occurs in approximately 40% of GBM, leading to enhanced activation of the EGFR signaling pathway and promoting tumor growth. Although GBM mutations are stably maintained in GBM in vitro models, rapid loss of EGFR gene amplification is a common observation during cell culture. To maintain EGFR amplification in vitro, heterotopic GBM xenografts with elevated EGFR copy number were cultured under varying serum conditions and EGF concentrations...
2017: PloS One
https://www.readbyqxmd.com/read/28918472/a-reliable-protocol-for-colorimetric-determination-of-iron-oxide-nanoparticle-uptake-by-cells
#3
Daiana K Deda, Roberta M Cardoso, Mayara K Uchiyama, Christiane Pavani, Sergio H Toma, Mauricio S Baptista, Koiti Araki
Size, shape, and surface properties of superparamagnetic iron oxide nanoparticles (SPIONs) can influence their interaction with biological systems, particularly the incorporation by tumor cells and consequently the biological activity and efficiency in biomedical applications. Several strategies have been used to evaluate cellular uptake of SPIONs. While qualitative methods are generally based on microscopy techniques, quantitative assays are carried out by techniques such as inductively coupled plasma-mass spectrometry and flow cytometry...
September 16, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28917808/single-drop-gold-nanoparticles-for-headspace-microextraction-and-colorimetric-assay-of-mercury-ii-in-environmental-waters
#4
Tesfaye Tolessa, Zhi-Qiang Tan, Yong-Guang Yin, Jing-Fu Liu
A novel headspace colorimetric nanosensor strategy for specific detection of Hg(II) was developed based upon analyte induced etching and amalgamation of gold nanoparticles (AuNPs). The Hg(II) was first selectively reduced to its volatile form, Hg(0), by stannous chloride (SnCl2) through chemical cold vapor generation (CVG) reaction. Then, the Hg(0) was headspace extracted into 37μL thioglycolic acid functionalized AuNP aqueous suspension containing 10% methanol as extractant and simultaneously reacted with AuNPs through the strong metallophilic Hg-Au interaction, resulting in a red-to-blue color change...
January 1, 2018: Talanta
https://www.readbyqxmd.com/read/28911344/comparison-of-spectrophotometric-methods-for-the-determination-of-copper-in-sugar-cane-spirit
#5
Sarah Adriana R Soares, Silvânio Silvério L Costa, Rennan Geovanny O Araujo, Leonardo Sena Gomes Teixeira, Alailson Falcão Dantas
Three spectrophotometric methods were developed for the determination of copper (Cu) in sugar cane spirit using the chromogenic reagents neocuproine, cuprizone, and bathocuproine. Experimental conditions, such as reagent concentration, reducer concentration, pH, buffer concentration, the order of addition of reagents, and the stability of the complexes, were optimized. The work range was established from 1.0 to 10.0 µg/mL, with correlation coefficients of >0.999 for all three optimized methods. The methods were evaluated regarding accuracy by addition and recovery tests at five concentration levels, and the obtained recoveries ranged from 91 to 105% (n = 3)...
September 15, 2017: Journal of AOAC International
https://www.readbyqxmd.com/read/28904817/ihc-based-subcellular-quantification-provides-new-insights-into-prognostic-relevance-of-flip-and-procaspase-8-in-non-small-cell-lung-cancer
#6
Ryan A Hutchinson, Helen G Coleman, Kathy Gately, Vincent Young, Siobhan Nicholson, Robert Cummins, Elaine Kay, Sean O Hynes, Philip D Dunne, Seedevi Senevirathne, Peter W Hamilton, Darragh G McArt, Daniel B Longley
In this study, we developed an image analysis algorithm for quantification of two potential apoptotic biomarkers in non-small-cell lung cancer (NSCLC): FLIP and procaspase-8. Immunohistochemical expression of FLIP and procaspase-8 in 184 NSCLC tumors were assessed. Individual patient cores were segmented and classified as tumor and stroma using the Definiens Tissue Studio. Subsequently, chromogenic expression of each biomarker was measured separately in the nucleus and cytoplasm and reported as a quantitative histological score...
2017: Cell Death Discovery
https://www.readbyqxmd.com/read/28903986/correction-for-perry-a-decade-of-development-of-chromogenic-culture-media-for-clinical-microbiology-in-an-era-of-molecular-diagnostics
#7
John D Perry
No abstract text is available yet for this article.
October 2017: Clinical Microbiology Reviews
https://www.readbyqxmd.com/read/28898981/performance-of-the-cobas-mrsa-sa-test-for-simultaneous-detection-of-methicillin-susceptible-and-methicillin-resistant-staphylococcus-aureus-from-nasal-swabs
#8
Lance R Peterson, Christopher W Woods, Thomas E Davis, Zi-Xuam Wang, Stephen A Young, John C Osiecki, Michael A Lewinski, Oliver Liesenfeld
Objectives: Health care-associated methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus aureus (SA) infections are continuing problems. Rapidly determining the MRSA colonization status of a patient facilitates practice to reduce spread of MRSA clinical disease. Sensitive detection of all SA prior to surgery, followed by decolonization, can significantly reduce postoperative infection from this pathogen. Our goal was to validate a new automated assay for this testing. Methods: We compared performance of the cobas MRSA/SA Test on the cobas 4800 System to direct and enriched chromogenic culture using nasal swabs collected from patients at six United States sites...
August 1, 2017: American Journal of Clinical Pathology
https://www.readbyqxmd.com/read/28892622/gold-loaded-nanoporous-ferric-oxide-nanocubes-with-peroxidase-mimicking-activity-for-electrocatalytic-and-colorimetric-detection-of-autoantibody
#9
Mostafa Kamal Masud, Sharda Yadav, Md Nazmul Islam, Nam-Trung Nguyen, Carlos Salomon, Richard Kline, Hatem R Alamri, Zeid Abdullah Alothman, Yusuke Yamauchi, Md Shahriar A Hossain, Muhammad J A Shiddiky
Enzyme-mimicking activity of iron oxide based nanostructures have provided a significant advantage in developing advanced molecular sensors for biomedical and environmental applications. Herein, we introduce the horseradish peroxidase (HRP)-like activity of gold loaded nanoporous ferric oxide nanocubes (Au-NPFe2O3NC) for the development of a molecular sensor with enhanced electrocatalytic and colorimetric (naked-eye) detection of autoantibodies. The results showed that Au-NPFe2O3NC exhibits enhanced peroxidase-like activity towards the catalytic oxidation of 3,3',5,5'-tertamethylbenzidine (TMB) in the presence of H2O2 at room temperature (25°C) and follows the typical Michaelis-Menten kinetics...
September 11, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28891289/in-situ-fluorogenic-and-chromogenic-reactions-for-the-sensitive-dual-readout-assay-of-tyrosinase-activity
#10
Jiahui Zhao, Xingfu Bao, Shuang Wang, Shasha Lu, Jian Sun, Xiurong Yang
As a well-known copper-containing oxidase, tyrosinase has been anticipated to serve as the biomarker of skin diseases. We describe here an exquisite label-free fluorescent and colorimetric dual-readout assay of its activity, inspired by the specific oxidation ability of monophenolamine substrates to catecholamines and a unique fluorogenic reaction between resorcinol and catecholamines. By employing commercially available tyramine as the model substrate (dopamine as the product), it is found that the tyrosinase-incubated tyramine solution exhibits obvious pale yellow with intense blue fluorescence in the presence of resorcinol and O2, where the absorbance and fluorescence intensity are directly related to the concentration of added tyrosinase (i...
September 22, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28889376/immunohistochemical-analysis-of-urothelial-carcinoma-tissues-for-proliferation-and-differentiation-markers
#11
Michael Rose, Nadine T Gaisa
Immunohistochemistry is a standard method in histopathology and enables the localized detection of proteins in histological tissue sections. Specific antibodies are bound to cellular antigens, captured by secondary antibodies or polymers, linked to enzymes and visualized by chromogenic substrates. Here, we describe an automated staining technique for larger slide batches as well as a manual protocol for only few slides using a polymer technique. We focus on differentiation markers, measures of cell proliferation, and therapeutic targets in benign and malignant urothelial tissues...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28888219/factor-xa-inhibition-by-rivaroxaban-in-the-trough-steady-state-can-significantly-reduce-thrombin-generation
#12
Shigeo Horinaka, Rie Sugawara, Yutaka Yonezawa, Toshihiko Ishimitsu
AIMS: The aim of the present study was to demonstrate evidence of reduced thrombin generation at the trough plasma rivaroxaban concentration. METHODS: A single-centre, prospective, non-randomised, drug intervention, self-controlled study was conducted in 51 anticoagulation therapy-naïve patients with non-valvular atrial fibrillation. Plasma rivaroxaban concentration was measured by liquid chromatography tandem mass spectrometry (LC-MS/MS) and anti-factor Xa chromogenic assay...
September 9, 2017: British Journal of Clinical Pharmacology
https://www.readbyqxmd.com/read/28888037/myc-drives-overexpression-of-telomerase-rna-htr-terc-in-prostate-cancer
#13
Javier A Baena-Del Valle, Qizhi Zheng, David M Esopi, Michael Rubenstein, Gretchen K Hubbard, Maria C Moncaliano, Andrew Hruszkewycz, Ajay Vaghasia, Srinivasan Yegnasubramanian, Sarah J Wheelan, Alan K Meeker, Christopher M Heaphy, Mindy K Graham, Angelo M De Marzo
Telomerase consists of at least two essential elements, an RNA component hTR or TERC that contains the template for telomere DNA addition, and a catalytic reverse transcriptase (TERT). While expression of TERT has been considered the key rate limiting component for telomerase activity, increasing evidence suggests an important role for the regulation of TERC in telomere maintenance and perhaps other functions in human cancer. By using three orthogonal methods including RNAseq, RT-qPCR, and an analytically validated chromogenic RNA in situ hybridization assay, we report consistent overexpression of TERC in prostate cancer...
September 9, 2017: Journal of Pathology
https://www.readbyqxmd.com/read/28886505/a-dual-responsive-colorimetric-and-fluorescent-chemosensor-based-on-diketopyrrolopyrrole-derivative-for-naked-eye-detection-of-fe-3-and-its-practical-application
#14
Shanshan Zhang, Tao Sun, Dejun Xiao, Fang Yuan, Tianduo Li, Enhua Wang, Haixia Liu, Qingfen Niu
A novel dual-responsive colorimetric and fluorescent chemosensor L based on diketopyrrolopyrrole derivative for Fe(3+) detection was designed and synthesized. In presence of Fe(3+), sensor L displayed strong colorimetric response as amaranth to rose pink and significant fluorescence enhancement and chromogenic change, which served as a naked-eye indicator by an obvious color change from purple to red. The binding constant for L-Fe(3+) complex was found as 2.4×10(4) with the lower detection limit of 14.3nM...
September 5, 2017: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/28882017/utility-of-rapid-microbiological-techniques-for-the-diagnosis-of-severe-infections
#15
REVIEW
E Cercenado
Rapid diagnostic microbiological techniques and antimicrobial susceptibility testing are necessary for early and adequate treatment. The utility of old (Gram stain, antigen detection, direct antimicrobial susceptibility testing, chromogenic media) and new techniques (molecular assays, MALDI-TOF) is summarized for the rapid diagnosis of bacteraemia and fungaemia, catheter-related bloodstream infections, pneumonia, meningitis, skin and soft-tissue infections, urinary tract infection, Clostridium difficile infection, viral infections, and tuberculosis...
September 2017: Revista Española de Quimioterapia: Publicación Oficial de la Sociedad Española de Quimioterapia
https://www.readbyqxmd.com/read/28873396/a-mouse-mastitis-model-to-study-the-effects-of-the-intramammary-infusion-of-a-food-grade-lactococcus-lactis-strain
#16
Cristina Camperio, Federica Armas, Elena Biasibetti, Paolo Frassanito, Carlo Giovannelli, Liliana Spuria, Claudia D'Agostino, Sabrina Tait, Maria Teresa Capucchio, Cinzia Marianelli
Lactococcus lactis is one of the most important microorganisms in the dairy industry and has "generally recognized as safe" (GRAS) status. L. lactis belongs to the group of lactic acid bacteria (LAB) and is encountered in a wide range of environments. Recently, the use of the intramammary infusion of a live culture of LAB has been investigated as a new antibiotic alternative for treating mastitis in dairy ruminants. Controversial results are described in literature regarding its efficacy and safety. In this study we conducted in-depth investigation of the mammary gland immune response induced by intramammary inoculum of a live culture of L...
2017: PloS One
https://www.readbyqxmd.com/read/28870730/mrsa-decolonization-control-comparing-a-single-day-swabbing-regimen-with-an-established-three-day-protocol
#17
Hagen Frickmann, Norbert Georg Schwarz, Andreas Hahn, Alicja Ludyga, Philipp Warnke, Andreas Podbielski
OBJECTIVES: Success of MRSA decolonization procedures is usually verified by control swabs of the colonized body region. This prospective controlled study compared a single-day regimen with a well-established 3-day scheme for noninferiority and adherence to the testing scheme. METHODS: Two sampling schemes for screening MRSA patients of a single study cohort at a German tertiary hospital 2 days after decolonization were compared regarding their ability to identify MRSA colonization in throat or nose...
September 1, 2017: Clinical Microbiology and Infection
https://www.readbyqxmd.com/read/28869002/mycobacterium-eburneum-sp-nov-a-non-chromogenic-fast-growing-strain-isolated-from-sputum
#18
Imen Nouioui, Lorena Carro, Kanae Teramoto, José M Igual, Marlen Jando, Maria Del Carmen Montero-Calasanz, Iain Sutcliffe, Vartul Sangal, Michael Goodfellow, Hans-Peter Klenk
A polyphasic study was undertaken to establish the taxonomic position of a non-chromogenic, rapidly growing Mycobacterium strain that had been isolated from sputum. The strain, CECT 8775T, has chemotaxonomic and cultural properties consistent with its classification in the genus Mycobacterium and was distinguished from the type strains of closely related mycobacterial species, notably from Mycobacterium paraense DSM 46749T, its nearest phylogenetic neighbour, based on 16S rRNA, hsp65 and rpoB gene sequence data...
September 2017: International Journal of Systematic and Evolutionary Microbiology
https://www.readbyqxmd.com/read/28866201/high-performance-peroxidase-mimics-for-rapid-colorimetric-detection-of-h2o2-and-glucose-derived-from-perylene-diimides-functionalized-co3o4-nanoparticles
#19
Yanan Ding, Miaomiao Chen, Kaili Wu, Mingxing Chen, Lifang Sun, Zhenxue Liu, Zhiqiang Shi, Qingyun Liu
N,N'-di-caboxy methyl perylene diimides (PDI), as one of the most promising functional materials in optional chemosensing, was first used to combine with Co3O4 nanoparticles through a facile two-step hydrothermal method and obtain the PDI functionalized Co3O4 nanocomposites (PDI-Co3O4 NCs). PDI-Co3O4 NCs were characterized by a series of technical analysis including transmission electron microscope (TEM), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR), respectively. The experimental results indicated that the as-prepared PDI-Co3O4 NCs possessed the higher peroxidase-like activity than that of Co3O4 nanoparticles without PDI, and could rapidly catalyze oxidation reaction of the chromogenic substrate TMB in the presence of H2O2 to a blue product (oxTMB) observed by the naked eye...
November 1, 2017: Materials Science & Engineering. C, Materials for Biological Applications
https://www.readbyqxmd.com/read/28856794/rapid-and-sensitive-colorimetric-detection-of-ascorbic-acid-in-food-based-on-the-intrinsic-oxidase-like-activity-of-mno2-nanosheets
#20
Liuying He, Feiyang Wang, Ying Chen, Yueying Liu
In this paper, we report a colorimetric sensor for the rapid, selective detection of ascorbic acid (AA) in aqueous solutions. Single-layered MnO2 nanosheets were established as an artificial oxidase; consequently colorless 3,3´,5,5´-tetramethylbenzidine (TMB) was oxidized to a blue product (oxTMB), with increase in absorbance at 650 nm. The absorbance of the reaction system decreased after introduction AA, which reduced MnO2 into Mn(2+) . Under optimum conditions, a detection limit of 62.81 nM for AA in aqueous solutions could be achieved...
August 30, 2017: Luminescence: the Journal of Biological and Chemical Luminescence
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