Abstract:
PURPOSE:The aim of this work was to develop a quantitative, flow cytometric method for tracking the endolysosomal escape of a fluorescently labelled saporin toxin. METHODS:Flow cytometric measurements of fluorescent pulse width and height were used to track the endocytic uptake into Daudi cells of a fluorescently labelled saporin toxin and the saporin based immunotoxin, OKT10-SAP. Subsequently, measurement of changes in pulse width were used to investigate the effect of a triterpenoid saponin on the endolysosomal escape of internalised toxin into the cytosol. Live cell confocal microscopy was used to validate the flow cytometry data. RESULTS:Increased endolysosomal escape of saporin and OKT10-SAP was observed by confocal microscopy in cells treated with saponin. Fluorescent pulse width measurements were also able to detect and quantify escape more sensitively than confocal microscopy. Saponin induced endolysosomal escape could be abrogated by treatment with chloroquine, an inhibitor of endolysosomal acidification. Chloroquine abrogation of escape was also mirrored by a concomitant abrogation of cytotoxicity. CONCLUSIONS:Poor endolysosomal escape is often a rate limiting step for the cytosolic delivery of protein toxins and other macromolecules. Pulse width analysis offers a simple method to semi-quantify the endolysosomal escape of this and similar molecules into the cytosol.
journal_name
Pharm Resjournal_title
Pharmaceutical researchauthors
Wensley HJ,Johnston DA,Smith WS,Holmes SE,Flavell SU,Flavell DJdoi
10.1007/s11095-019-2725-1subject
Has Abstractpub_date
2019-12-23 00:00:00pages
16issue
1eissn
0724-8741issn
1573-904Xpii
10.1007/s11095-019-2725-1journal_volume
37pub_type
杂志文章abstract:PURPOSE:Information on genetic variants that affect the pharmacokinetics and pharmacodynamics (PK/PD) of drugs in different populations from Mexico is still an ongoing endeavor. Here, we investigate allele frequencies on pharmacogenetic targets in Mexican Mestizos and Natives from three different States and its associa...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-016-1990-5
更新日期:2016-11-01 00:00:00
abstract:PURPOSE:This study aimed to investigate the impact of the size of X-ray iodinated contrast agent in nano-emulsions, on their toxicity and fate in vivo. METHODS:A new compound, triiodobenzoate cholecalciferol, was synthetized, formulated as nano-emulsions, and followed after i.v. administration in mice by X-ray imaging...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-015-1813-0
更新日期:2016-03-01 00:00:00
abstract:PURPOSE:Our previous studies showed that the mRNA level of human organic anion transporter (hOAT) 3 in the kidney was correlated with the rate of elimination of an anionic antibiotic cefazolin. However, the correlation coefficient was not so high. In the present study, therefore, we enrolled more patients to examine wh...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-005-8383-5
更新日期:2005-12-01 00:00:00
abstract:PURPOSE:The objective of this study was to develop and evaluate a pulsatile drug delivery system based on an impermeable capsule body filled with drug and an erodible plug placed in the opening of the capsule body. METHODS:The erodible plugs were either prepared by direct compression followed by placing the tablets in...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1011940718534
更新日期:1998-03-01 00:00:00
abstract:PURPOSE:Multidrug and toxin extrusion proteins (MATEs) are multispecific organic cation transporters mediating the efflux of various cationic drugs into the urine. The present study aimed at identifying endogenous compounds in human plasma and urine specimens as biomarkers to evaluate drug interactions involving MATEs ...
journal_title:Pharmaceutical research
pub_type: 临床试验,杂志文章
doi:10.1007/s11095-013-1144-y
更新日期:2014-01-01 00:00:00
abstract:PURPOSE:To compare the physical stability of amorphous molecular level solid dispersions of nifedipine and felodipine, in the presence of poly(vinylpyrrolidone) (PVP) and small amounts of moisture. METHODS:Thin amorphous films of nifedipine and felodipine and amorphous molecular level solid dispersions with PVP were s...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-007-9420-3
更新日期:2008-03-01 00:00:00
abstract:PURPOSE:To develop an enzymatically-controlled pulsatile drug release system based on an impermeable capsule body, which contains the drug and is closed by an erodible pectin/pectinase-plug. METHODS:The plug was prepared by direct compression of pectin and pectinase in different ratios. In addition to the disintegrati...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1018959327311
更新日期:1999-09-01 00:00:00
abstract::The effect of γ-cyclodextrin and its four derivatives on the solubility of progesterone in phosphate buffer pH 7.4 was investigated. γ-Cyclodextrin forms a complex precipitating from solution at low cyclodextrin concentrations. No precipitation of complexes was observed with the γ-cyclodextrin derivatives. This change...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/A:1016301919928
更新日期:1985-11-01 00:00:00
abstract:PURPOSE:A plasmid-based gene expression system was complexed with protective, interactive, and non-condensing (PINC) polymer system and administered with Medi-Jector, a needle-free injection device (NFID), to achieve high and sustained levels of antigen-specific antibodies in blood circulation. METHODS:Human growth ho...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1018834305079
更新日期:1999-06-01 00:00:00
abstract:PURPOSE:The gastric mucoadhesive property of tetracycline-sucralfate acidic complex (CO) was evaluated by using a novel method in vitro to compare with the in vivo test. The mucoadhesive mechanism of the acidic complex was also studied. METHODS:The gastric mucosa removed from a rat was placed covering the end of a plu...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/B:PHAM.0000019293.57927.7f
更新日期:2004-03-01 00:00:00
abstract:PURPOSE:The objective of this study was to develop and characterize long-circulating, biodegradable, and biocompatible nanoparticulate formulation as an intracellular delivery vehicle. METHODS:Poly(ethylene glycol) (PEG)-modified gelatin was synthesized by reacting Type-B gelatin with PEG-epoxide. The nanoparticles, p...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1016486910719
更新日期:2002-07-01 00:00:00
abstract::Molecular modeling has been used to study the complexation between alpha, beta, or gamma-cyclodextrin (CD) and a group of amine compounds having the diphenylmethyl functionality. The computer program SYBYL 5.3 and the Tripos force field (version 5.2) were used for all the calculations. Three-dimensional structures of ...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1015816215751
更新日期:1991-10-01 00:00:00
abstract:PURPOSE:The addition of high amounts of calcium remains a pharmaceutical concern due to its precipitation with phosphate in total parenteral nutrient (TPN) admixtures, compromising also the stability of the lipid emulsion. MATERIALS AND METHODS:Calcium-phosphate solubility was compared when using binary PN solutions v...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-008-9671-7
更新日期:2008-11-01 00:00:00
abstract::The mechanism of release from sustained-release adinazolam mesylate tablets was assessed by the Higuchi equation and by analysis of drug release profiles through 60% released using the Peppas equation. Computed values of the diffusional exponent, n, ranged from 0.59 to 0.66. Values of n in this range are consistent wi...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1015834114359
更新日期:1991-12-01 00:00:00
abstract:PURPOSE:To determine whether LHRH-receptor is expressed in Calu-3, a human bronchial epithelial cell line, and to further determine whether this receptor plays a role in the transport of deslorelin, an LHRH agonist. METHODS:Using cultured monolayers of Calu-3 grown at air-interface, the presence and localization of LH...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/b:pham.0000029294.70707.74
更新日期:2004-06-01 00:00:00
abstract::Since ancient times, natural products, herbs and spices have been used for preventing several diseases, including cancer. The term chemoprevention was coined in the late 1970s and referred to the prevention of cancer by selective use of phytochemicals or their analogs. The field utilizes experimental carcinogenesis mo...
journal_title:Pharmaceutical research
pub_type: 历史文章,杂志文章,评审
doi:10.1007/s11095-010-0085-y
更新日期:2010-06-01 00:00:00
abstract:PURPOSE:This study was conducted to investigate in vivo the impact of interferon-alpha (IFN)-alpha on P-glycoprotein (P-gp) activity in rats by studying how its administration modifies the bioavailability of digoxin, a fairly pure P-gp substrate. METHODS:Human recombinant IFN-alpha was given to rats (n = 5-7 per group...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-005-7415-5
更新日期:2005-11-01 00:00:00
abstract:PURPOSE:A novel photo-activated targeted chemotherapy was developed by photochemical internalization (PCI) of glutathione-sensitive polymeric micelles incorporating camptothecin (CPT) prepared from thiolated CPT (CPT-DP) and thiolated poly(ethylene glycol)-b-poly(glutamic acid) (PEG-b-P(Glu-DP)) METHODS: PEG-b-P(Glu-DP...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-008-9712-2
更新日期:2009-01-01 00:00:00
abstract::The technique of Affinity capillary electrophoresis (ACE) is a useful tool to characterize complexation and partition equilibria. A wide range of applications in pharmaceutics has been presented: the determination of pKA values, of association constants between drugs and cyclodextrins or amphiphilic compounds as well ...
journal_title:Pharmaceutical research
pub_type: 杂志文章,评审
doi:
更新日期:1999-11-01 00:00:00
abstract:PURPOSE:The work aims at investigating the correlation of water sorption potential with different measures of molecular mobility in an annealed amorphous model compound (trehalose). METHODS:Amorphous trehalose, prepared by freeze-drying, was annealed at 100°C (17°C < T (g)) for up to 120 h. Global mo...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-012-0910-6
更新日期:2013-03-01 00:00:00
abstract:PURPOSE:Microphysiological systems (MPS), also known as "organs-on-chips" or "tissue chips," leverage recent advances in cell biology, tissue engineering, and microfabrication to create in vitro models of human organs and tissues. These systems offer promising solutions for modeling human physiology and disease in vitr...
journal_title:Pharmaceutical research
pub_type: 杂志文章,评审
doi:10.1007/s11095-019-2742-0
更新日期:2019-12-17 00:00:00
abstract:PURPOSE:To evaluate different dissolution testing methods and subsequently develop a simple to perform but reproducible and discriminating dissolution technique for inhalative powders. METHODS:From a dry powder a fraction of aerosolized particles with an aerodynamic particle size below 5 μm was collected on regenerate...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-012-0744-2
更新日期:2012-08-01 00:00:00
abstract:PURPOSE:The objective of this study was to investigate the possible influence of probenecid on morphine transport across the blood-brain barrier (BBB) in rats. METHODS:Microdialysis probes, calibrated using retrodialysis by drug, were placed into the striatum and jugular vein of seven Sprague-Dawley rats. Morphine was...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1023250900462
更新日期:2003-04-01 00:00:00
abstract:PURPOSE:The objective of this investigation is to develop a rational design of Osteotropic Drug Delivery System (ODDS), which we have proposed as a novel method for drug delivery to the skeleton via bisphosphonic prodrug, based on the relationship between physicochemical and pharmacokinetic properties of bisphosphonate...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1011033326980
更新日期:2001-05-01 00:00:00
abstract:PURPOSE:To investigate, for the first time, the performance of a dry powder inhaler (DPI, Aerolizer(®)) in the case of a model drug (i.e. albuterol sulphate) formulated with spray dried mannitol carrier particles with homogeneous shape and solid-state form but different sizes. METHODS:Spray dried mannitol (SDM) partic...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-013-1132-2
更新日期:2014-01-01 00:00:00
abstract::Human serum albumin (HSA) was encapsulated in a 50:50 copolymer of DL-lactide/glycolide in the form of microspheres. These microspheres were used as a model formulation to study the feasibility of controlling the release of large proteins over a 20- to 30-day period. We show that HSA can be successfully incorporated i...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1015948829632
更新日期:1990-11-01 00:00:00
abstract::Chimeric L6 is a mouse-human monoclonal antibody specific for tumor cell-associated antigens. The factors affecting the physical and chemical stability of chimeric L6 were assessed at elevated temperatures (30-60 degrees C) and by multiple freezing and thawing. Three routes of degradation were observed: chemical degra...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1018948901599
更新日期:1994-05-01 00:00:00
abstract:PURPOSE:To study the structure and function of the intestinal H+/ peptide transporter PET1, we compared its amino acid sequence with those of related transporters belonging to the oligopeptide transporter family PTR, and with more distant transporter families. METHODS:We have developed a new approach to the sequence a...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1012070726480
更新日期:1997-04-01 00:00:00
abstract:PURPOSE:Adjuvants are required to ensure the efficacy of subunit vaccines. Incorporating molecular immunopotentiators within particles could overcome drawbacks of molecular adjuvants (such as solubility and toxicity), and improve adjuvanticity of particles, achieving stronger adjuvant activity. Aim of this study is to ...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1007/s11095-015-1666-6
更新日期:2015-09-01 00:00:00
abstract::Recently a new in vivo approach in man, using a regional intestinal perfusion technique, has been developed. The perfusion tube consists of a multichannel tube with two inflatable balloons, which are placed 10 cm apart. The tube is introduced orally and the time required for insertion and positioning of the tube is ap...
journal_title:Pharmaceutical research
pub_type: 杂志文章
doi:10.1023/a:1015888813741
更新日期:1992-10-01 00:00:00