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- Strömberg, Anette, et al.
(författare)
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Manipulating the genetic identity and biochemical surface properties of individual cells with electric-field-induced fusion
- 2000
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - Stanford Univ, Dept Chem, Stanford, CA 94305 USA. Gothenburg Univ, Dept Chem, S-41296 Gothenburg, Sweden. Sahlgrens Univ Hosp, Inst Clin Neurosci, Dept Neurol, S-41345 Gothenburg, Sweden. : NATL ACAD SCIENCES. - 0027-8424 .- 1091-6490. ; 97:1, s. 7-11
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Tidskriftsartikel (refereegranskat)abstract
- A method for cell-cell and cell-liposome fusion at the single-cell level is described. Individual cells or liposomes were first selected and manipulated either by optical trapping or by adhesion to a micromanipulator-controlled ultramicroelectrode. Spatially selective fusion of the cell-cell or cell-liposome pair was achieved by the application of a highly focused electric field through a pair of 5-mu m o.d. carbon-fiber ultramicroelectrodes. The ability to fuse together single cells opens new possibilities in the manipulation of the genetic and cellular makeup of individual cells in a controlled manner, In the study of cellular networks, for example, the alteration of the biochemical identity of a selected cell can have a profound effect on the behavior of the entire network. Fusion of a single liposome with a target cell allows the introduction of the liposomal content into the cell interior as well as the addition of lipids and membrane proteins onto the cell surface. This cell-liposome fusion represents an approach to the manipulation of the cytoplasmic contents and surface properties of single cells. As an example, we have introduced a membrane protein (gamma-glutamyltransferase) reconstituted in liposomes into the cell plasma membrane.
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| 3. |
- Wilhelmsson, Marcus, 1974, et al.
(författare)
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Genetic screening using the colour change of a PNA-DNA hybrid-binding cyanine dye
- 2002
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Ingår i: Nucleic Acids Research. - : Oxford University Press (OUP). - 0305-1048 .- 1362-4962. ; 30:2
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Tidskriftsartikel (refereegranskat)abstract
- As the relationship between human genes and various malfunctions and diseases becomes revealed at an ever-increasing pace, the need arises for the development of rapid genetic screening methods for diagnostic purposes. Genetic diseases show great diversity. Some are caused by a few characteristic localised mutations, while others arise from a large number of variations. Hence, It. is unlikely that. a single, general diagnostic method that applies to all cases will ever exist. Instead, a combination of methods is frequently applied. Here we propose the use of a dramatic colour change that a cyanine ye, 3,3'-diethylthiadicarbocyanine, displays upon binding to DNA-PNA duplexes. This method could become an inexpensive, fast and simple genetic screening test by visual inspection, with no need for complicated equipment. Our results demonstrate that this diagnostic method may be sufficiently sensitive to discriminate between even a fully complementary and a single mutation DNA sequence.
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| 4. |
- Wilson, C F, et al.
(författare)
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Nanoengineered structures for holding and manipulating liposomes and cells
- 2001
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Ingår i: Analytical Chemistry. - Stanford Univ, Dept Chem, Stanford, CA 94305 USA. Univ Gothenburg, Dept Chem, SE-41296 Gothenburg, Sweden. : AMER CHEMICAL SOC. - 0003-2700 .- 1520-6882. ; 73:4, s. 787-791
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Tidskriftsartikel (refereegranskat)abstract
- We describe the fabrication of nanoengineered holding pipets with concave seating surfaces and fine pressure control. These pipets were shown to exhibit exceptional stability in capturing, transporting, and releasing single cells and liposomes 1-12 mum in diameter, which opens previously inaccessible avenues of research. Three specific examples demonstrated the, utility and versatility of this manipulation system. In the first, carboxyrhodaminie was selectively incorporated into individual cells by electroporation, after which nearly all the medium (hundreds of microliters) surrounding the docked and tagged cells was rapidly exchanged (in seconds) and the cells were subsequently probed by laser-induced fluorescence (LIF). In the second study, a single liposome containing carboxyrhodamine was transported to a dye-free solution using a transfer pipet, docked to a holding pipet, and held firmly during physical agitation and interrogation by LIF. In the third study, pairs of liposomes were positioned between two microelectrodes, held in contact, and selectively electrofused and the resulting liposomes undocked intact.
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| 5. |
- Zare, Fariba, 1972, et al.
(författare)
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Arthritogenic properties of double-stranded (viral) RNA
- 2004
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Ingår i: J Immunol. - 0022-1767. ; 172:9, s. 5656-63
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Tidskriftsartikel (refereegranskat)abstract
- Viral infections often lead to arthralgias and overt arthritic states. The inflammatogenic compound of the viruses giving rise to such an outcome has to date not been identified. Because expression of dsRNA is a common feature of all viruses, we decided to analyze whether this property leads to the induction of arthritis. Histological signs of arthritis were evident already on day 3 following intra-articular administration of dsRNA. Arthritis was characterized by infiltration of macrophages into synovial tissue. It was not dependent on acquired immune responses because SCID mice also raised joint inflammation. NF-kappa B was activated upon in vitro exposure to dsRNA, indicating its role in the induction/progression of arthritis. Importantly, we found that dsRNA arthritis was triggered through IL-1R signaling because mice being deficient for this molecule were unable to develop joint inflammation. Although dsRNA is typically recognized by Toll-like receptor 3, Toll-like receptor 3 knockout mice developed arthritis, indicating that some other receptors are instrumental in the inducing of inflammation. Our results from in vitro experiments indicate that proinflammatory cytokines and chemokines stimulating monocyte influx were readily triggered in response to stimulation with dsRNA. These findings demonstrate that viral dsRNA is clearly arthritogenic. Importantly, macrophages and their products play an important role in the development of arthritis triggered by dsRNA.
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