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Intracellular pH flow cytometry

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https://www.readbyqxmd.com/read/29737834/ph-controlled-liposomes-for-enhanced-cell-penetration-in-tumor-environment
#1
Michela Barattin, Andrea Mattarei, Anna Balasso, Cristina Paradisi, Laura Cantù, Elena Del Favero, Tapani Viitala, Francesca Mastrotto, Paolo Caliceti, Stefano Salmaso
An innovative pH-switchable colloidal system that can be exploited for site-selective anticancer drug delivery has been generated by liposomes decoration with a new novel synthetic non-peptidic oligo-Arginine cell-penetration enhancer (CPE) and a quenching PEGylated counterpart that detaches from the vesicle surface under the acidic conditions of tumors. The CPE module (Arg4-DAG) is formed by four arginine units conjugated to a first generation (G1) 2,2-bis(hydroxymethyl)propionic acid (bis-MPA)/2,2-bis(aminomethyl)propionic acid (bis-AMPA) polyester dendron terminating with 1,2-distearoyl-3-azidopropane for liposome bilayer insertion...
May 8, 2018: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/29712729/conservation-of-intracellular-pathogenic-strategy-among-distantly-related-cryptococcal-species
#2
Joudeh B Freij, Man Shun Fu, Carlos M De Leon Rodriguez, Amanda Dziedzic, Anne E Jedlicka, Quigly Dragotakes, Diego C P Rossi, Eric H Jung, Carolina Coelho, Arturo Casadevall
The genus Cryptococcus includes several pathogenic species for humans. Until recently, the two major pathogenic species were recognized as C. neoformans (Cn) and C. gattii (Cg). We compared the interaction of murine macrophages with three Cg species complex strains (WM179, R265, and WM161, representing molecular types VGI, VGIIa, and VGIII, respectively) and one Cn species complex strain (H99, molecular type VNI) to ascertain similarities and differences in the yeast intracellular pathogenic strategy. Parameters analyzed include non-lytic exocytosis frequency, phagolysosomal pH, intracellular capsular growth, phagolysosomal membrane permeabilization, and macrophage transcriptional response, assessed using time-lapse microscopy, fluorescence microscopy, flow cytometry and gene expression microarray analysis...
April 30, 2018: Infection and Immunity
https://www.readbyqxmd.com/read/29690762/purification-characterization-and-mode-of-action-of-plantaricin-gz1-27-a-novel-bacteriocin-against-bacillus-cereus
#3
Hechao Du, Jie Yang, Xiaohong Lu, Zhaoxin Lu, Xiaomei Bie, Haizhen Zhao, Chong Zhang, Fengxia Lu
Bacillus cereus is an opportunistic pathogen that causes foodborne diseases. We isolated a novel bacteriocin, designated plantaricin GZ1-27, and elucidated its mode of action against B. cereus. Plantaricin GZ1-27 was purified using ammonium sulfate precipitation, gel-filtration chromatography, and RP-HPLC. MALDI-TOF/MS revealed that its molecular mass was 975 Da, and Q-TOF-MS/MS analysis predicted the amino acid sequence as VSGPAGPPGTH. Plantaricin GZ1-27 showed thermostability and pH stability. The antibacterial mechanism was investigated using flow cytometry, confocal laser-scanning microscopy, scanning and transmission electron microscopy, and RT-PCR, which revealed that GZ1-27 increased cell membrane permeability, triggered K+ leakage and pore formation, damaged cell membrane integrity, altered cell morphology and intracellular organization, and reduced the expression of genes related to cytotoxin production, peptidoglycan synthesis, and cell division...
May 1, 2018: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/29579564/2-2-dithiodisuccinic-acid-stabilized-polyion-complex-micelles-for-ph-and-reduction-dual-responsive-drug-delivery
#4
Ying Du, Wei Yan, Hua Lian, Chenyang Xiang, Lijie Duan, Chunsheng Xiao
In this work, a new kind of polyion complex (PIC) micelles with pH and reduction dual responsiveness was developed for effective intracellular drug delivery. The PIC micelles can be readily prepared by mixing a polycationic block polymer, methoxy poly(ethylene glycol)-b-poly(l-lysine) (mPEG-PLL), with a small molecule polyacid, 2, 2'-dithiodisuccinic acid (DTS) in aqueous media. The resultant PIC micelles are of uniform spherical shapes with hydrodynamic radii ranging from 65 to 75 nm based on different feeding ratios of mPEG-PLL and DTS...
March 15, 2018: Journal of Colloid and Interface Science
https://www.readbyqxmd.com/read/29575108/coumarin-bodipy-hybridization-for-ratiometric-sensing-of-intracellular-polarity-oscillation
#5
Weijiang He, Yang Bai, Xiangchao Shi, Yuncong Chen, Chengcheng Zhu, Yang Jiao, Zhong Han, Zijian Guo
With the different polarity response, coumarin and BODIPY were hybridized to construct polarity fluorescent sensors, CBDP and iso-CBDP, to overcome the disadvantages of solvatochromic sensors in ratiometric polarity sensing. Only CBDP displayed that the emission ratio (Icou/IBDP, coumarin to BODIPY emissions) increased with an exponential dependence on medium relative permittivity in a wide polarity range. This sensing ability of CBDP was not interfered by medium pH, viscosity, and most intracellular species especially reactive oxygen, nitrogen, and sulfur species...
March 25, 2018: Chemistry: a European Journal
https://www.readbyqxmd.com/read/29497295/polyaspartic-acid-anchored-mesoporous-silica-nanoparticles-for-ph-responsive-doxorubicin-release
#6
Abdul Hakeem, Fouzia Zahid, Guiting Zhan, Ping Yi, Hai Yang, Lu Gan, Xiangliang Yang
Background: Nanotechnology-based drug delivery systems exhibit promising therapeutic efficacy in cancer chemotherapy. However, ideal nano drug carriers are supposed to be sufficiently internalized into cancer cells and then release therapeutic cargoes in response to certain intracellular stimuli, which has never been an easy task to achieve. Objective: This study is to design mesoporous silica nanoparticles (MSNs)-based pH-responsive nano drug delivery system that is effectively internalized into cancer cells and then release drug in response to lysosomal/endosomal acidified environment...
2018: International Journal of Nanomedicine
https://www.readbyqxmd.com/read/29487850/regulating-netosis-increasing-ph-promotes-nadph-oxidase-dependent-netosis
#7
Meraj A Khan, Lijy M Philip, Guillaume Cheung, Shawn Vadakepeedika, Hartmut Grasemann, Neil Sweezey, Nades Palaniyar
Neutrophils migrating from the blood (pH 7.35-7.45) into the surrounding tissues encounter changes in extracellular pH (pHe ) conditions. Upon activation of NADPH oxidase 2 (Nox), neutrophils generate large amounts of H+ ions reducing the intracellular pH (pHi ). Nevertheless, how extracellular pH regulates neutrophil extracellular trap (NET) formation (NETosis) is not clearly established. We hypothesized that increasing pH increases Nox-mediated production of reactive oxygen species (ROS) and neutrophil protease activity, stimulating NETosis...
2018: Frontiers in Medicine
https://www.readbyqxmd.com/read/29477643/paclitaxel-controlled-delivery-using-a-ph-responsive-functional-aunp-block-copolymer-vesicular-nanocarrier-composite-system
#8
Athina Liaskoni, Athina Angelopoulou, Efstathia Voulgari, Maria-Teodora Popescu, Constantinos Tsitsilianis, Konstantinos Avgoustakis
Paclitaxel (PTX)-loaded gold nanoparticles functionalized with mercaptooctanoic acid (MOA) and folic acid (FA) (AuMOA-FA) were encapsulated within pH-sensitive poly(2-vinylpyridine)-b-poly(ethylene oxide) (P2VP-PEO) vesicles with the aim to develop a more selective injectable nano-formulation for PTX, lacking the side effects of the conventional PTX delivery system. The size of the resulting composite vesicles was lower than 200 nm, i.e. it is suitable for tumor targeting applications taking advantage of the enhanced permeability and retention (EPR) effect...
May 30, 2018: European Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/29445584/acid-sensing-ion-channel-1a-regulates-fate-of-rat-nucleus-pulposus-cells-in-acid-stimulus-through-endoplasmic-reticulum-stress
#9
Zhi-Yang Xie, Lu Chen, Cong Zhang, Lei Liu, Feng Wang, Feng Cai, Xiao-Hu Wang, Rui Shi, Arjun Sinkemani, Hao-Min Yu, Xin Hong, Xiao-Tao Wu
Acid-sensing ion channel 1a (ASIC1a) participates in human intervertebral disc degeneration (IVDD) and regulates the destiny of nucleus pulposus cells (NPCs) in acid stimulus. However, the mechanism of ASIC1a activation and its downstream pathway remain unclear. Endoplasmic reticulum (ER) stress also participates in the acid-induced apoptosis of NPCs. The main purpose of this study was to investigate whether there is any connection between ASIC1a and ER stress in an acid-induced nucleus pulposus degeneration model...
2018: BioResearch Open Access
https://www.readbyqxmd.com/read/29343959/antitumor-effect-of-a-new-nano-vector-with-mirna-135a-on-malignant-glioma
#10
Chaofeng Liang, Weitong Sun, Haiyong He, Baoyu Zhang, Cong Ling, Bocheng Wang, Tengchao Huang, Bo Hou, Ying Guo
Introduction: MiR-135a is found to selectively induce apoptosis in glioma cell but not in normal neurons and glial cells. However, low transfection efficacy limits its application in vivo as other miRNAs. We prepared a new kind of nano-vector based on polyethylene glycol methyl ether (mPEG) and hyper-branched polyethylenimine (hy-PEI) in order to improve the miRNA delivery system into the glioma cells. Methods: The mPEG-g-PEI/miR-135a was constructed and detected by 1H NMR and FTIR analyses...
2018: International Journal of Nanomedicine
https://www.readbyqxmd.com/read/29284258/ph-sensitive-coiled-coil-peptide-cross-linked-hyaluronic-acid-nanogels-synthesis-and-targeted-intracellular-protein-delivery-to-cd44-positive-cancer-cells
#11
Lingling Ding, Yu Jiang, Jian Zhang, Harm-Anton Klok, Zhiyuan Zhong
The clinical translation of protein drugs that act intracellularly is limited by the absence of safe and efficient intracellular protein delivery vehicles. Here, pH-sensitive coiled-coil peptide-cross-linked hyaluronic acid nanogels (HA-cNGs) were designed and investigated for targeted intracellular protein delivery to CD44 overexpressing MCF-7 breast cancer cells. HA-cNGs were obtained with a small size of 176 nm from an equivalent mixture of hyaluronic acid conjugates with GY(EIAALEK)3 GC (E3) and GY(KIAALKE)3 GC (K3) peptides, respectively, at pH 7...
February 12, 2018: Biomacromolecules
https://www.readbyqxmd.com/read/29188718/polyplex-micelles-with-phenylboronate-gluconamide-cross-linking-in-the-core-exerting-promoted-gene-transfection-through-spatiotemporal-responsivity-to-intracellular-ph-and-atp-concentration
#12
Naoto Yoshinaga, Takehiko Ishii, Mitsuru Naito, Taisuke Endo, Satoshi Uchida, Horacio Cabral, Kensuke Osada, Kazunori Kataoka
Polyplexes as gene delivery carriers require integrated functionalities to modulate intracellular trafficking for efficient gene transfection. Herein, we developed plasmid DNA (pDNA)-loaded polyplex micelles (PMs) from poly(ethylene glycol)-based block catiomers derivatized with 4-carboxy-3-fluorophenylboronic acid (FPBA) and d-gluconamide to form pH- and ATP-responsive cross-linking in the core. These PMs exhibited robustness in the extracellular milieu and smooth endosomal escape after cellular uptake, and they facilitated pDNA decondensation triggered by increased ATP concentration inside of the cell...
December 27, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/29186618/semi-automatized-segmentation-method-using-image-based-flow-cytometry-to-study-sperm-physiology-the-case-of-capacitation-induced-tyrosine-phosphorylation
#13
Arturo Matamoros-Volante, Ayelen Moreno-Irusta, Paulina Torres-Rodriguez, Laura Giojalas, María G Gervasi, Pablo E Visconti, Claudia L Treviño
STUDY QUESTION: Is image-based flow cytometry a useful tool to study intracellular events in human sperm such as protein tyrosine phosphorylation or signaling processes? SUMMARY ANSWER: Image-based flow cytometry is a powerful tool to study intracellular events in a relevant number of sperm cells, which enables a robust statistical analysis providing spatial resolution in terms of the specific subcellular localization of the labeling. WHAT IS KNOWN ALREADY: Sperm capacitation is required for fertilization...
February 1, 2018: Molecular Human Reproduction
https://www.readbyqxmd.com/read/29135959/intratumoral-delivery-of-doxorubicin-on-folate-conjugated-graphene-oxide-by-in-situ-forming-thermo-sensitive-hydrogel-for-breast-cancer-therapy
#14
Yi Teng Fong, Chih-Hao Chen, Jyh-Ping Chen
By taking advantage of the pH-sensitive drug release property of graphene oxide (GO) after intracellular uptake, we prepared folic acid (FA)-conjugated GO (GOFA) for targeted delivery of the chemotherapeutic drug doxorubicin (DOX). GOFA-DOX was further encapsulated in an injectable in-situ forming thermo-sensitive hyaluronic acid-chitosan- g -poly( N -isopropylacrylamide) (HACPN) hydrogel for intratumoral delivery of DOX. As the degradation time of HACPN could be extended up to 3 weeks, intratumoral delivery of GOFA-DOX/HACPN could provide controlled and targeted delivery of DOX through slow degradation HACPN and subsequent cellular uptake of released GOFA-DOX by tumor cells through interactions of GOFA with folate receptors on the tumor cell's surface...
November 14, 2017: Nanomaterials
https://www.readbyqxmd.com/read/29109030/layer-by-layer-assembly-of-graphene-oxide-on-thermosensitive-liposomes-for-photo-chemotherapy
#15
Mohadeseh Hashemi, Meisam Omidi, Bharadwaj Muralidharan, Lobat Tayebi, Matthew J Herpin, Mohammad Ali Mohagheghi, Javad Mohammadi, Hugh D C Smyth, Thomas E Milner
Stimuli responsive polyelectrolyte nanoparticles have been developed for chemo-photothermal destruction of breast cancer cells. This novel system, called layer by layer Lipo-graph (LBL Lipo-graph), is composed of alternate layers of graphene oxide (GO) and graphene oxide conjugated poly (l-lysine) (GO-PLL) deposited on cationic liposomes encapsulating doxorubicin. Various concentrations of GO and GO-PLL were examined and the optimal LBL Lipo-graph was found to have a particle size of 267.9 ± 13 nm, zeta potential of +43...
January 2018: Acta Biomaterialia
https://www.readbyqxmd.com/read/29039643/exploring-the-ph-sensitivity-of-poly-allylamine-phosphate-supramolecular-nanocarriers-for-intracellular-sirna-delivery
#16
Patrizia Andreozzi, Eleftheria Diamanti, Karen Rapp Py-Daniel, Paolin Rocio Cáceres-Vélez, Chiara Martinelli, Nikolaos Politakos, Ane Escobar, Marco Muzi-Falconi, Ricardo Azevedo, Sergio E Moya
Silencing RNA (siRNA) technologies emerge as a promising therapeutic tool for the treatment of multiple diseases. An ideal nanocarrier (NC) for siRNAs should be stable at physiological pH and release siRNAs in acidic endosomal pH, fulfilling siRNA delivery only inside cells. Here, we show a novel application of polyamine phosphate NCs (PANs) based on their capacity to load negatively charged nucleic acids and their pH stability. PANs are fabricated by complexation of phosphate anions from phosphate buffer solution (PB) with the amine groups of poly(allylamine) hydrochloride as carriers for siRNAs...
November 8, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28968600/the-h-k-atpase-inhibitor-sch-28080-inhibits-insulin-secretion-and-induces-cell-death-in-ins-1e-rat-insulinoma-cells
#17
Martin Jakab, Nina Ketterl, Johannes Fürst, Marlena Beyreis, Michael Kittl, Tobias Kiesslich, Cornelia Hauser-Kronberger, Martin Gaisberger, Markus Ritter
BACKGROUND/AIMS: Glucose-stimulated insulin secretion (GSIS) of pancreatic β-cells involves glucose uptake and metabolism, closure of KATP channels and depolarization of the cell membrane potential (Vmem), activation of voltage-activated Ca2+ currents (ICav) and influx of Ca2+, which eventually triggers hormone exocytosis. Beside this classical pathway, KATP-independent mechanisms such as changes in intracellular pH (pHi) or cell volume, which also affect β-cell viability, can elicit or modify insulin release...
2017: Cellular Physiology and Biochemistry
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
#18
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
#19
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...
October 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
#20
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...
November 1, 2017: Colloids and Surfaces. B, Biointerfaces
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