A comparison between different human hepatocyte models reveals profound differences in net glucose production, lipid composition and metabolism in vitro.
AuthorsBonanini F,Singh M,Yang H,Kurek D,Harms AC,Mardinoglu A,Hankemeier T
JournalExperimental cell research
PubMed ID38499143
Effects of an indole derivative on cell proliferation, transfection, and alternative splicing in production of lentiviral vectors by transient co-transfection.
AuthorsMier NC,Roper DK
JournalPloS one
PubMed ID38833479
Lentiviral vectors derived from human immunodeficiency virus type I are widely used to deliver functional gene copies to mammalian cells for research and gene therapies. Post-transcriptional splicing of lentiviral vector transgene in transduced host and transfected producer cells presents barriers to widespread application of lentiviral vector-based therapies. The present study ... More
Metabolism of 4 beta -hydroxycholesterol in humans.
'One of the major oxysterols in the human circulation is 4 beta-hydroxycholesterol formed from cholesterol by the drug-metabolizing enzyme cytochrome P450 3A4. Deuterium-labeled 4 beta-hydroxycholesterol was injected into two healthy volunteers, and the apparent half-life was found to be 64 and 60 h, respectively. We have determined earlier the half-lives ... More
Delivery of proteins into living cells by reversible membrane permeabilization with streptolysin-O.
AuthorsWalev I, Bhakdi SC, Hofmann F, Djonder N, Valeva A, Aktories K, Bhakdi S,
JournalProc Natl Acad Sci U S A
PubMed ID11248053
'The pore-forming toxin streptolysin O (SLO) can be used to reversibly permeabilize adherent and nonadherent cells, allowing delivery of molecules with up to 100 kDa mass to the cytosol. Using FITC-labeled albumin, 10(5)-10(6) molecules were estimated to be entrapped per cell. Repair of toxin lesions depended on Ca(2+)-calmodulin and on ... More
Use of human tissue explants to study human infectious agents.
AuthorsGrivel JC, Margolis L,
JournalNat Protoc
PubMed ID19197269
The study of human cell-cell and cell-pathogen interactions that occur in the context of complex tissue cytoarchitecture is critical for deciphering the mechanisms of many normal and pathogenic processes. This protocol describes methods for culturing and infecting explants of human tissues to study the pathogenesis of human infectious agents and ... More
Polycystin-1, the product of the polycystic kidney disease 1 gene, co-localizes with desmosomes in MDCK cells.
AuthorsScheffers MS, van der Bent P, Prins F, Spruit L, Breuning MH, Litvinov SV, de Heer E, Peters DJ,
JournalHum Mol Genet
PubMed ID11063733
Polycystin-1 is a novel protein predicted to be a large membrane-spanning glycoprotein with an extracellular N-terminus and an intracellular C-terminus, harboring several structural motifs. To study the subcellular localization, antibodies raised against various domains of polycystin-1 and against specific adhesion complex proteins were used for two-color immunofluorescence staining. In Madine ... More
Apoptosis induces efflux of the mitochondrial matrix enzyme deoxyguanosine kinase.
AuthorsJüllig M, Eriksson S,
JournalJ Biol Chem
PubMed ID11294860
Deoxyguanosine kinase (dGK) initiates the salvage of purine deoxynucleosides in mitochondria and is a key enzyme in mitochondrial DNA precursor synthesis. The active form of the enzyme is a 60-kDa protein normally located in the mitochondrial matrix. Here we describe the subcellular distribution of dGK during apoptosis in human epithelial ... More
Purification, Cloning, and Characterization of Nek8, a Novel NIMA-related Kinase, and Its Candidate Substrate Bicd2.
Authors Holland Pamela M; Milne Alison; Garka Kirsten; Johnson Richard S; Willis Cynthia; Sims John E; Rauch Charles T; Bird Timothy A; Virca G Duke;
JournalJ Biol Chem
PubMed ID11864968
We describe the isolation, cloning, and characterization of human Nek8, a new mammalian NIMA-related kinase, and its candidate substrate Bicd2. Nek8 was isolated as a beta-casein kinase activity in rabbit lung and has an N-terminal catalytic domain homologous to the Nek family of protein kinases. Nek8 also contains a central ... More
Malignant effusions and immunogenic tumour-derived exosomes.
BACKGROUND: Exosomes derived from tumours are small vesicles released in vitro by tumour cell lines in culture supernatants. To assess the role of these exosomes in vivo, we examined malignant effusions for their presence. We also investigated whether these exosomes could induce production of tumour-specific T cells when pulsed with ... More