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Intracellular pH flow cytometry

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https://www.readbyqxmd.com/read/28882160/intrinsic-fluorescence-of-the-clinically-approved-multikinase-inhibitor-nintedanib-reveals-lysosomal-sequestration-as-resistance-mechanism-in-fgfr-driven-lung-cancer
#1
Bernhard Englinger, Sebastian Kallus, Julia Senkiv, Daniela Heilos, Lisa Gabler, Sushilla van Schoonhoven, Alessio Terenzi, Patrick Moser, Christine Pirker, Gerald Timelthaler, Walter Jäger, Christian R Kowol, Petra Heffeter, Michael Grusch, Walter Berger
BACKGROUND: Studying the intracellular distribution of pharmacological agents, including anticancer compounds, is of central importance in biomedical research. It constitutes a prerequisite for a better understanding of the molecular mechanisms underlying drug action and resistance development. Hyperactivated fibroblast growth factor receptors (FGFRs) constitute a promising therapy target in several types of malignancies including lung cancer. The clinically approved small-molecule FGFR inhibitor nintedanib exerts strong cytotoxicity in FGFR-driven lung cancer cells...
September 7, 2017: Journal of Experimental & Clinical Cancer Research: CR
https://www.readbyqxmd.com/read/28800409/cytotoxic-effects-of-the-proton-pump-inhibitor-omeprazole-on-the-non-target-marine-microalga-tetraselmis-suecica
#2
Marta Seoane, Marta Esperanza, Ángeles Cid
Omeprazole (OMP) is one of the most commonly used drugs for the treatment of gastro-intestinal disorders. Although it is daily consumed in high quantities and commonly detected in waters worldwide, relatively little is known about its ecotoxicity. The aim of this study was to evaluate the potential acute toxicity of increasing concentrations of OMP on the marine microalga Tetraselmis suecica analysing several cytotoxicity biomarkers by flow cytometry after 24h of exposure. Results showed that OMP caused a decrease in growth and autofluorescence, an increase in cellular volume and intracellular complexity, hyperpolarization of cytoplasmic and mitochondrial membranes and intracellular acidification...
August 2, 2017: Aquatic Toxicology
https://www.readbyqxmd.com/read/28797971/modified-pamam-dendrimer-with-4-carbomethoxypyrrolidone-surface-groups-its-uptake-efflux-and-location-in-a-cell
#3
Anna Janaszewska, Maciej Studzian, Johannes F Petersen, Mario Ficker, Valentina Paolucci, Jørn B Christensen, Donald A Tomalia, Barbara Klajnert-Maculewicz
Traditional amine terminated PAMAM dendrimers may be readily surface engineered by a facile one-pot conversion with dialkyl itaconate esters into 4-carbomethoxypyrrolidone terminated PAMAM (G=0-4) dendrimers. These terminated dendrimers are uniquely characterized by exhibiting blue fluorescence emissions (λexc=370nm, λmaxem=440nm). Thanks to this property they can be directly analyzed by confocal microscopy and flow cytometry without additional fluorescence labeling, treatment of dendrimers with chemicals or adjusting pH...
July 29, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28778000/non-proinflammatory-and-responsive-nanoplatforms-for-targeted-treatment-of-atherosclerosis
#4
Yin Dou, Yue Chen, Xiangjun Zhang, Xiaoqiu Xu, Yidan Chen, Jiawei Guo, Dinglin Zhang, Ruibing Wang, Xiaohui Li, Jianxiang Zhang
Atherosclerosis is the leading cause of many fatal cardiovascular and cerebrovascular diseases. Whereas nanomedicines are promising for targeted therapy of atherosclerosis, great challenges remain in development of effective, safe, and translational nanotherapies for its treatment. Herein we hypothesize that non-proinflammatory nanomaterials sensitive to low pH or high reactive oxygen species (ROS) may serve as effective platforms for triggerable delivery of anti-atherosclerotic therapeutics in cellular and tissue microenvironments of inflammation...
July 29, 2017: Biomaterials
https://www.readbyqxmd.com/read/28752896/metabolic-reprogramming-by-hif-1-activation-enhances-survivability-of-human-adipose-derived-stem-cells-in-ischaemic-microenvironments
#5
Chang Chen, Qi Tang, Yan Zhang, Minjia Dai, Yichen Jiang, Hang Wang, Mei Yu, Wei Jing, Weidong Tian
OBJECTIVES: Poor cell survival severely limits the beneficial effect of adipose-derived stem cell (ADSC)-based therapy for disease treatment and tissue regeneration, which might be caused by the attenuated level of hypoxia-inducible factor-1 (HIF-1) in these cells after having been cultured in 21% ambient oxygen in vitro for weeks. In this study, we explored the role of pre-incubation in dimethyloxalylglycine (DMOG, HIF-1 activator) in the survivability of human ADSCs in a simulated ischaemic microenvironment in vitro and in vivo...
October 2017: Cell Proliferation
https://www.readbyqxmd.com/read/28750328/inactivation-efficiency-of-plasmid-encoded-antibiotic-resistance-genes-during-water-treatment-with-chlorine-uv-and-uv-h2o2
#6
Younggun Yoon, Hay Jung Chung, Doris Yoong Wen Di, Michael C Dodd, Hor-Gil Hur, Yunho Lee
This study assessed the inactivation efficiency of plasmid-encoded antibiotic resistance genes (ARGs) both in extracellular form (e-ARG) and present within Escherichia coli (intracellular form, i-ARG) during water treatment with chlorine, UV (254 nm), and UV/H2O2. A quantitative real-time PCR (qPCR) method was used to quantify the ARG damage to amp(R) (850 bp) and kan(R) (806 bp) amplicons, both of which are located in the pUC4K plasmid. The plate count and flow cytometry methods were also used to determine the bacterial inactivation parameters, such as culturability and membrane damage, respectively...
June 22, 2017: Water Research
https://www.readbyqxmd.com/read/28732161/elegant-ph-responsive-nanovehicle-for-drug-delivery-based-on-triazine-dendrimer-modified-magnetic-nanoparticles
#7
Amir Landarani-Isfahani, Majid Moghadam, Shima Mohammadi, Maryam Royvaran, Naimeh Moshtael-Arani, Saghar Rezaei, Shahram Tangestaninejad, Valiollah Mirkhani, Iraj Mohammadpoor-Baltork
Owing to properties of magnetic nanoparticles and elegant three-dimensional macromolecule architectural features, dendrimeric structures have been investigated as nanoscale drug delivery systems. In this work, a novel magnetic nanocarrier, generation two (G2) triazine dendrimer modified Fe3O4@SiO2 magnetic nanoparticles (MNP-G2), was designed, fabricated, and characterized by Fourier transform infrared (FT-IR), thermal gravimetric analysis (TGA), vibrating sample magnetometer (VSM), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), and dynamic light scattering (DLS)...
August 15, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/28704808/enhanced-reactive-oxygen-species-production-acidic-cytosolic-ph-and-upregulated-na-h-exchanger-nhe-in-dicer-deficient-cd4-t-cells
#8
Yogesh Singh, Yuetao Zhou, Shaqiu Zhang, Khalid N M Abdelazeem, Bernat Elvira, Madhuri S Salker, Florian Lang
BACKGROUND: MicroRNAs (miRNAs) negatively regulate gene expression at a post-transcriptional level. Dicer, a cytoplasmic RNase III enzyme, is required for the maturation of miRNAs from precursor miRNAs. Dicer, therefore, is a critical enzyme involved in the biogenesis and processing of miRNAs. Several biological processes are controlled by miRNAs, including the regulation of T cell development and function. T cells generate reactive oxygen species (ROS) with parallel H+ extrusion accomplished by the Na+/H+-exchanger 1 (NHE1)...
July 14, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28685162/in-vivo-indicators-of-cytoplasmic-vacuolar-and-extracellular-ph-using-phluorin2-in-candida-albicans
#9
Hélène Tournu, Arturo Luna-Tapia, Brian M Peters, Glen E Palmer
Environmental or chemically induced stresses often trigger physiological responses that regulate intracellular pH. As such, the capacity to detect pH changes in real time and within live cells is of fundamental importance to essentially all aspects of biology. In this respect, pHluorin, a pH-sensitive variant of green fluorescent protein, has provided an invaluable tool to detect such responses. Here, we report the adaptation of pHluorin2 (PHL2), a substantially brighter variant of pHluorin, for use with the human fungal pathogen Candida albicans...
July 2017: MSphere
https://www.readbyqxmd.com/read/28671589/theranostic-liposome-nanoparticle-hybrids-for-drug-delivery-and-bioimaging
#10
Muharrem Seleci, Didem Ag Seleci, Thomas Scheper, Frank Stahl
Advanced theranostic nanomedicine is a multifunctional approach which combines the diagnosis and effective therapy of diseased tissues. Here, we investigated the preparation, characterization and in vitro evaluation of theranostic liposomes. As is known, liposome-quantum dot (L-QD) hybrid vesicles are promising nanoconstructs for cell imaging and liposomal-topotecan (L-TPT) enhances the efficiency of TPT by providing protection against systemic clearance and allowing extended time for it to accumulate in tumors...
July 2, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28576550/comparison-of-different-zeolite-framework-types-as-carriers-for-the-oral-delivery-of-the-poorly-soluble-drug-indomethacin
#11
Christina Karavasili, Elsa P Amanatiadou, Eleni Kontogiannidou, Georgios K Eleftheriadis, Nikolaos Bouropoulos, Eleni Pavlidou, Ioanna Kontopoulou, Ioannis S Vizirianakis, Dimitrios G Fatouros
Microporous zeolites of distinct framework types, textural properties and crystal morphologies namely BEA, ZSM and NaX, have been employed as carriers to assess their effect on modulating the dissolution behavior of a BCS II model drug (indomethacin). Preparation of the loaded carriers via the incipient wetness method induced significant drug amorphization for the BEA and NaX samples, as well as high drug payloads. The stability of the amorphous drug content was retained after stressing test evaluation of the porous carriers...
August 7, 2017: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/28484514/increased-lignocellulosic-inhibitor-tolerance-of-saccharomyces-cerevisiae-cell-populations-in-early-stationary-phase
#12
Venkatachalam Narayanan, Jenny Schelin, Marie Gorwa-Grauslund, Ed Wj van Niel, Magnus Carlquist
BACKGROUND: Production of second-generation bioethanol and other bulk chemicals by yeast fermentation requires cells that tolerate inhibitory lignocellulosic compounds at low pH. Saccharomyces cerevisiae displays high plasticity with regard to inhibitor tolerance, and adaptation of cell populations to process conditions is essential for reaching efficient and robust fermentations. RESULTS: In this study, we assessed responses of isogenic yeast cell populations in different physiological states to combinations of acetic acid, vanillin and furfural at low pH...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/28482576/folate-decorated-pegylated-triblock-copolymer-as-a-ph-reduction-dual-responsive-nanovehicle-for-targeted-intracellular-co-delivery-of-doxorubicin-and-bcl-2-sirna
#13
Aili Suo, Junmin Qian, Minghui Xu, Weijun Xu, Yaping Zhang, Yu Yao
Co-delivery of chemotherapeutic drug and small interfering RNA (siRNA) within a single nanovehicle has emerged as a promising combination therapy approach to treating cancers because of their synergistic effect. Nanocarrier delivery systems with low cytotoxicity and high efficiency are needed for such a purpose. In this study, a novel folate-conjugated PEGylated cationic triblock copolymer, poly(acrylhydrazine)-block-poly(3-dimethylaminopropyl methacrylamide)-block-poly(acrylhydrazine) (PAH-b-PDMAPMA-b-PAH), was synthesized and evaluated as a stimuli-sensitive vehicle for the targeted co-delivery of doxorubicin (DOX) and Bcl-2 siRNA into breast cancer MCF-7 cells...
July 1, 2017: Materials Science & Engineering. C, Materials for Biological Applications
https://www.readbyqxmd.com/read/28479046/a-triple-fluorophore-labeled-nucleic-acid-ph-nanosensor-to-investigate-non-viral-gene-delivery
#14
David R Wilson, Denis Routkevitch, Yuan Rui, Arman Mosenia, Karl J Wahlin, Alfredo Quinones-Hinojosa, Donald J Zack, Jordan J Green
There is a need for new tools to better quantify intracellular delivery barriers in high-throughput and high-content ways. Here, we synthesized a triple-fluorophore-labeled nucleic acid pH nanosensor for measuring intracellular pH of exogenous DNA at specific time points in a high-throughput manner by flow cytometry following non-viral transfection. By including two pH-sensitive fluorophores and one pH-insensitive fluorophore in the nanosensor, detection of pH was possible over the full physiological range...
July 5, 2017: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/28342376/label-free-silicon-nanodots-featured-ratiometric-fluorescent-aptasensor-for-lysosomal-imaging-and-ph-measurement
#15
Yanan Zhang, Shan Guo, Shibo Cheng, Xinghu Ji, Zhike He
The homeostasis of lysosomal pH is crucial in cell physiology. Developing small fluorescent nanosensors for lysosome imaging and ratiometric measurement of pH is highly demanded yet challenging. Herein, a pH-sensitive fluorescein tagged aptamer AS1411 has been utilized to covalently modify the label-free fluorescent silicon nanodots via a crosslinker for construction of a ratiometric pH biosensor. The established aptasensor exhibits the advantages of ultrasmall size, hypotoxicity, excellent pH reversibility and good photostability, which favors its application in an intracellular environment...
March 22, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28261191/heterogeneity-between-and-within-strains-of-lactobacillus-brevis-exposed-to-beer-compounds
#16
Yu Zhao, Susanne Knøchel, Henrik Siegumfeldt
This study attempted to investigate the physiological response of six Lactobacillus brevis strains to hop stress, with and without the addition of Mn(2+) or ethanol. Based on the use of different fluorescent probes, cell viability and intracellular pH (pHi) were assessed by fluorescence microscopy combined with flow cytometry, at the single cell level. The combined approach was faster than the traditional colony based method, but also provided additional information about population heterogeneity with regard to membrane damage and cell size reduction, when exposed to hop compounds...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28244372/action-mechanisms-involved-in-the-bioprotective-effect-of-lactobacillus-harbinensis-k-v9-3-1-np-against-yarrowia-lipolytica-in-fermented-milk
#17
Sophie Mieszkin, Nolwenn Hymery, Stella Debaets, Emmanuel Coton, Gwenaelle Le Blay, Florence Valence, Jérôme Mounier
The use of lactic acid bacteria (LAB) as bioprotective cultures can be an alternative to chemical preservatives or antibiotic to prevent fungal spoilage in dairy products. Among antifungal LAB, Lactobacillus harbinensis K.V9.3.1Np showed a remarkable antifungal activity for the bioprotection of fermented milk without modifying their organoleptic properties (Delavenne et al., 2015). The aim of the present study was to elucidate the action mechanism of this bioprotective strain against the spoilage yeast Yarrowia lipolytica...
May 2, 2017: International Journal of Food Microbiology
https://www.readbyqxmd.com/read/28224106/impact-of-high-dose-vitamin-c-on-platelet-function
#18
Bassem M Mohammed, Kimberly W Sanford, Bernard J Fisher, Erika J Martin, Daniel Contaifer, Urszula Osinska Warncke, Dayanjan S Wijesinghe, Charles E Chalfant, Donald F Brophy, Alpha A Fowler Iii, Ramesh Natarajan
AIM: To examine the effect of high doses of vitamin C (VitC) on ex vivo human platelets (PLTs). METHODS: Platelet concentrates collected for therapeutic or prophylactic transfusions were exposed to: (1) normal saline (control); (2) 0.3 mmol/L VitC (Lo VitC); or (3) 3 mmol/L VitC (Hi VitC, final concentrations) and stored appropriately. The VitC additive was preservative-free buffered ascorbic acid in water, pH 5.5 to 7.0, adjusted with sodium bicarbonate and sodium hydroxide...
February 4, 2017: World Journal of Critical Care Medicine
https://www.readbyqxmd.com/read/28217898/metal-induced-changes-in-trivalent-chromium-resistant-alcaligenes-faecalis-vitsim2
#19
Shiny C Matilda, Chittibabu Shanthi
The changes induced in bacterial strains under stress conditions provide an insight into metal resistance strategies. Trivalent chromium resistant bacterium were isolated and identified by 16S rRNA gene sequencing and designated as Alcaligenes faecalis VITSIM2. The growth pattern was monitored. The organism also showed resistance to copper, cadmium, and certain antibiotics. The differentially expressed proteins in SDS PAGE were identified by mass spectrometry as flagellin and 50S ribosomal L36 protein. The morphological changes were identified by scanning electron microscopy...
May 2017: Journal of Basic Microbiology
https://www.readbyqxmd.com/read/28186394/mechanisms-of-ph-sensitivity-and-cellular-internalization-of-peoz-b-pla-micelles-with-varied-hydrophilic-hydrophobic-ratios-and-intracellular-trafficking-routes-and-fate-of-the-copolymer
#20
Dishi Wang, Yanxia Zhou, Xinru Li, Xiaoyou Qu, Yunqiang Deng, Ziqi Wang, Chuyu He, Yang Zou, Yiguang Jin, Yan Liu
pH-responsive polymeric micelles have shown promise for the targeted and intracellular delivery of antitumor agents. The present study aimed to elucidate the possible mechanisms of pH-sensitivity and cellular internalization of PEOz-b-PLA micelles in detail, further unravel the effect of hydrophilic/hydrophobic ratio of the micelles on their cellular internalization, and examine the intracellular trafficking routes and fate of PEOz-b-PLA after internalization of the micelles. The results of variations in the size and Zeta potential of PEOz-b-PLA micelles and cross-sectional area of PEOz-b-PLA molecules with pH values suggested that electrostatic repulsion between PEOz chains resulting from ionization of the tertiary amide groups along PEOz chain at pH lower than its pKa was responsible for pH-sensitivity of PEOz-b-PLA micelles...
March 1, 2017: ACS Applied Materials & Interfaces
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