Intracellular trafficking of Clostridium perfringens iota-toxin b.
AuthorsNagahama M, Umezaki M, Tashiro R, Oda M, Kobayashi K, Shibutani M, Takagishi T, Ishidoh K, Fukuda M, Sakurai J,
JournalInfect Immun
PubMed ID22825447
'Clostridium perfringens iota-toxin is composed of an enzymatic component (Ia) and a binding component (Ib). Ib binds to a cell surface receptor, undergoes oligomerization in lipid rafts, and binds Ia. The resulting complex is then endocytosed. Here, we show the intracellular trafficking of iota-toxin. After the binding of the Ib ... More
Oxidative stress protection by exogenous delivery of rhHsp70 chaperone to the retinal pigment epithelium (RPE), a possible therapeutic strategy against RPE degeneration.
AuthorsSubrizi A, Toropainen E, Ramsay E, Airaksinen AJ, Kaarniranta K, Urtti A,
Journal
PubMed ID25030185
'To measure the cytoprotective effects of rhHsp70 against oxidative stress and study its cellular uptake, intracellular and intraocular distribution in the retinal pigment epithelium. Human retinal pigment epithelial cells (ARPE-19) were pre-treated with rhHsp70 for 24 h, 48 h, and 72 h before being exposed to 1.25 mM hydrogen peroxide. Non-treated cells served as ... More
Epithelial cell mitochondrial dysfunction and PINK1 are induced by transforming growth factor-beta1 in pulmonary fibrosis.
AuthorsPatel AS, Song JW, Chu SG, Mizumura K, Osorio JC, Shi Y, El-Chemaly S, Lee CG, Rosas IO, Elias JA, Choi AM, Morse D,
Journal
PubMed ID25785991
'Epithelial cell death is a major contributor to fibrogenesis in the lung. In this study, we sought to determine the function of mitochondria and their clearance (mitophagy) in alveolar epithelial cell death and fibrosis. We studied markers of mitochondrial injury and the mitophagy marker, PTEN-induced putative kinase 1 (PINK1), in ... More
Visualizing the endocytosis of phenylephrine in living cells by quantum dot-based tracking.
AuthorsMa J, Wu L, Hou Z, Song Y, Wang L, Jiang W,
Journal
PubMed ID24855959
'To study the intracellular receptor-drug transportation, a fluorescent probe consisting of phenylephrine-polyethylene glycol-quantum dots conjugate was employed to track endocytosis process of phenylephrine in living cells. This type of movement was studied by continuously filming fluorescent images in the same cell. We also calculated the movement parameters, and divided the ... More
Dynamic colocalization microscopy to characterize intracellular trafficking of nanomedicines.
AuthorsVercauteren D, Deschout H, Remaut K, Engbersen JF, Jones AT, Demeester J, De Smedt SC, Braeckmans K,
JournalACS Nano
PubMed ID21923168
'To gain a better understanding of intracellular processing of nanomedicines, we employed quantitative live-cell fluorescence colocalization microscopy to study endosomal trafficking of polyplexes in retinal pigment epithelium cells. A new, dynamic colocalization algorithm was developed, based on particle tracking and trajectory correlation, allowing for spatiotemporal characterization of internalized polyplexes in ... More
Self-assembling dual component nanoparticles with endosomal escape capability.
AuthorsWong AS, Mann SK, Czuba E, Sahut A, Liu H, Suekama TC, Bickerton T, Johnston AP, Such GK,
Journal
PubMed ID25731820
'This study reports a novel nanoparticle system with simple and modular one-step assembly, which can respond intelligently to biologically relevant variations in pH. Importantly, these particles also show the ability to induce escape from the endosomal/lysosomal compartments of the cell, which is integral to the design of efficient polymeric delivery ... More
Polycations such as polyethylenimine (PEI) are used in many novel nonviral vector designs and there are continuous efforts to increase our mechanistic understanding of their interactions with cells. Even so, the mechanism of polyplex escape from the endosomal/lysosomal pathway after internalization is still elusive. The ... More
Inhibition of Ebola virus entry by a C-peptide targeted to endosomes.
AuthorsMiller EH, Harrison JS, Radoshitzky SR, Higgins CD, Chi X, Dong L, Kuhn JH, Bavari S, Lai JR, Chandran K,
JournalJ Biol Chem
PubMed ID21454542
Ebola virus (EboV) and Marburg virus (MarV) (filoviruses) are the causative agents of severe hemorrhagic fever. Infection begins with uptake of particles into cellular endosomes, where the viral envelope glycoprotein (GP) catalyzes fusion between the viral and host cell membranes. This fusion event is thought to involve conformational rearrangements of ... More
Inhibitors of intravesicular acidification protect against Shiga toxin in a pH-independent manner.
AuthorsDyve Lingelem AB, Bergan J, Sandvig K,
JournalTraffic
PubMed ID22132807
Shiga toxin inhibits protein synthesis after being transported from the cell surface to endosomes and retrogradely through the Golgi apparatus to the endoplasmic reticulum (ER) and into the cytosol. In this study, we have abolished proton gradients across internal membranes in different ways and investigated the effect on the various ... More
Covalent coupling of gum arabic onto superparamagnetic iron oxide nanoparticles for MRI cell labeling: physicochemical and in vitro characterization.
AuthorsPalma SI, Carvalho A, Silva J, Martins P, Marciello M, Fernandes AR, del Puerto Morales M, Roque AC,
Journal
PubMed ID25766788
Gum arabic (GA) is a hydrophilic composite polysaccharide derived from exudates of Acacia senegal and Acacia seyal trees. It is biocompatible, possesses emulsifying and stabilizing properties and has been explored as coating agent of nanomaterials for biomedical applications, namely magnetic nanoparticles (MNPs). Previous studies focused on the adsorption of GA ... More
Pathogenic LRRK2 mutations, through increased kinase activity, produce enlarged lysosomes with reduced degradative capacity and increase ATP13A2 expression.
AuthorsHenry AG, Aghamohammadzadeh S, Samaroo H, Chen Y, Mou K, Needle E, Hirst WD,
Journal
PubMed ID26251043
Lysosomal dysfunction plays a central role in the pathogenesis of several neurodegenerative disorders, including Parkinson's disease (PD). Several genes linked to genetic forms of PD, including leucine-rich repeat kinase 2 (LRRK2), functionally converge on the lysosomal system. While mutations in LRRK2 are commonly associated with autosomal-dominant PD, the physiological and ... More
The abcEDCBA-Encoded ABC Transporter and the virB Operon-Encoded Type IV Secretion System of Brucella ovis Are Critical for Intracellular Trafficking and Survival in Ovine Monocyte-Derived Macrophages.
AuthorsMacedo AA, Silva AP, Mol JP, Costa LF, Garcia LN, Araújo MS, Martins Filho OA, Paixão TA, Santos RL,
Journal
PubMed ID26366863
Brucella ovis infection is associated with epididymitis, orchitis and infertility in rams. Most of the information available on B. ovis and host cell interaction has been generated using murine macrophages or epithelial cell lines, but the interaction between B. ovis and primary ovine macrophages has not been studied. The aim ... More
PSMA-mediated endosome escape-accelerating polymeric micelles for targeted therapy of prostate cancer and the real time tracing of their intracellular trafficking.
AuthorsGao Y, Li Y, Li Y, Yuan L, Zhou Y, Li J, Zhao L, Zhang C, Li X, Liu Y,
Journal
PubMed ID25419788
The cytotoxicity of chemotherapeutic agents to healthy organs and drug resistance of tumor cells are believed to be the main obstacles to the successful cancer chemotherapy in the clinic. To ensure that anticancer drugs could be delivered to the tumor region, are quickly released from carriers in tumor cells and ... More
Effective endogenous gene silencing mediated by pH responsive peptides proceeds via multiple pathways.
AuthorsLam JK, Liang W, Lan Y, Chaudhuri P, Chow MY, Witt K, Kudsiova L, Mason AJ,
JournalJ Control Release
PubMed ID22138072
Cationic amphipathic histidine rich peptides possess high plasmid DNA and siRNA delivery capabilities. To further understand the pH responsive siRNA delivery process and evaluate the capabilities of such peptides we have investigated their ability to mediate specific silencing of endogenous GAPDH gene activity in MCF-7 and A549 cells and compared ... More
Engineering of blended nanoparticle platform for delivery of mitochondria-acting therapeutics.
AuthorsMarrache S, Dhar S,
JournalProc Natl Acad Sci U S A
PubMed ID22991470
Mitochondrial dysfunctions cause numerous human disorders. A platform technology based on biodegradable polymers for carrying bioactive molecules to the mitochondrial matrix could be of enormous potential benefit in treating mitochondrial diseases. Here we report a rationally designed mitochondria-targeted polymeric nanoparticle (NP) system and its optimization for efficient delivery of various ... More
Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell.
AuthorsSzaszák M, Steven P, Shima K, Orzekowsky-Schröder R, Hüttmann G, König IR, Solbach W, Rupp J,
JournalPLoS Pathog
PubMed ID21779161
Chlamydia trachomatis is an obligate intracellular bacterium that alternates between two metabolically different developmental forms. We performed fluorescence lifetime imaging (FLIM) of the metabolic coenzymes, reduced nicotinamide adenine dinucleotides [NAD(P)H], by two-photon microscopy for separate analysis of host and pathogen metabolism during intracellular chlamydial infections. NAD(P)H autofluorescence was detected inside ... More
Spreading of neurodegenerative pathology via neuron-to-neuron transmission of ß-amyloid.
Alzheimer's disease (AD) is the major cause of dementia. During the development of AD, neurofibrillary tangles progress in a fixed pattern, starting in the transentorhinal cortex followed by the hippocampus and cortical areas. In contrast, the deposition of ß-amyloid (Aß) plaques, which are the other histological hallmark of AD, does ... More
Baculovirus-mediated gene transfer into mammalian cells.
AuthorsBoyce FM, Bucher NL,
JournalProc Natl Acad Sci U S A
PubMed ID8637876
This paper describes the use of the baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) as a vector for gene delivery into mammalian cells. A modified AcMNPV virus was prepared that carried the Escherichia coli lacZ reporter gene under control of the Rous sarcoma virus promoter and mammalian RNA processing ... More
BacMam technology and its application to drug discovery.
AuthorsAmes RS, Kost TA, Condreay JP,
JournalExpert Opin Drug Discov
PubMed ID23488908
The recombinant baculovirus/insect cell system was firmly established as a leading method for recombinant protein production when a new potential use for these viruses was revealed in 1995. It was reported that engineered recombinant baculoviruses could deliver functional expression cassettes to mammalian cell types; a system which has come to ... More
Tools and techniques to measure mitophagy using fluorescence microscopy.
AuthorsDolman NJ, Chambers KM, Mandavilli B, Batchelor RH, Janes MS,
Journal
PubMed ID24121704
Mitophagy is a specialized form of autophagy that removes damaged mitochondria, thereby maintaining efficient cellular metabolism and reducing cellular stress caused by aberrant oxidative bursts. Deficits in mitophagy underlie several diseases, and a substantial body of research has elucidated key steps in the pathways that lead to and execute autophagic ... More
Cellular uptake of cyclotide MCoTI-I follows multiple endocytic pathways.
Cyclotides are plant-derived proteins that naturally exhibit various biological activities and whose unique cyclic structure makes them remarkably stable and resistant to denaturation or degradation. These attributes, among others, make them ideally suited for use as drug development tools. This study investigated the cellular uptake of cyclotide, MCoTI-I in live ... More
Targeted Delivery of Deoxycytidine Kinase to Her2-Positive Cells Enhances the Efficacy of the Nucleoside Analog Fludarabine.
AuthorsKoduvayur SP, Su Y, Kay BK, Lavie A,
JournalPLoS One
PubMed ID27280468
'Cytotoxic drugs, such as nucleoside analogs and toxins, commonly suffer from off-target effects. One approach to mitigate this problem is to deliver the cytotoxic drug selectively to the intended site. While for toxins this can be achieved by conjugating the cell-killing moiety to a targeting moiety, it is not an ... More
Immuno-potentiating pathway of HBsAg-HBIG immunogenic complex visualized.
AuthorsLiu H, Geng S, Wang B, Wu B, Xie X, Wang S, Zhong Y, Wang X, Qu D, Wen Y, Wang B,
JournalHum Vaccin Immunother
PubMed ID26618396
'Chronic viral hepatitis B (CHB) is a major global health problem. A therapeutic vaccine for CHB comprised of yeast-derived recombinant HBsAg-anti-HBs immunogenic complexes (YIC) has been devloped by us. A series of clinical trials has shown its therapeutic efficacy in decreasing HBV viral load and converting serum HBeAg-positive to anti-HBe-positive ... More
The cytochrome P450 inhibitor SKF-525A disrupts autophagy in primary rat hepatocytes.
AuthorsLuo Y, Yang X, Shi Q,
JournalChem Biol Interact
PubMed ID26964495
'The cytochrome P450 (CYP) inhibitor SKF-525A is commonly used to study drug metabolism and toxicity, particularly hepatotoxicity. By using Western blot and immunofluorescence staining, we unexpectedly found that SKF-525A at 2-20 µM caused remarkable accumulation of microtubule-associated protein light chain 3 II (LC3-II) in primary rat hepatocytes at 1, 4 and ... More
Direct Observation of a-Synuclein Amyloid Aggregates in Endocytic Vesicles of Neuroblastoma Cells.
Aggregation of a-synuclein has been linked to both familial and sporadic Parkinson's disease. Recent studies suggest that a-synuclein aggregates may spread from cell to cell and raise questions about the propagation of neurodegeneration. While continuous progress has been made characterizing a-synuclein aggregates in vitro, there is a lack of information ... More
Binding, internalization and fate of Huntingtin Exon1 fibrillar assemblies in mitotic and nonmitotic neuroblastoma cells.
The aggregation of Huntingtin (HTT) protein and of its moiety encoded by its Exon1 (HTTExon1) into fibrillar structures inside neurons is the molecular hallmark of Huntington's disease. Prion-like transmission of these aggregates between cells has been demonstrated. The cell-to-cell transmission mechanisms of these protein aggregates and the susceptibility of different ... More
Calcium phosphate-based organic-inorganic hybrid nanocarriers with pH-responsive on/off switch for photodynamic therapy.
AuthorsNomoto T, Fukushima S, Kumagai M, Miyazaki K, Inoue A, Mi P, Maeda Y, Toh K, Matsumoto Y, Morimoto Y, Kishimura A, Nishiyama N, Kataoka K,
JournalBiomater Sci
PubMed ID26971562
Photodynamic therapy (PDT) is a promising treatment modality for malignant tumors in a light-selective manner. To improve the PDT efficacy, numerous kinds of nanocarriers have been developed to deliver photosensitizers (PSs) selectively into the tumor through leaky tumor-associated vasculature. However, the corresponding prolonged retention of the nanocarrier in the bloodstream ... More