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| 2. |
- Hilson, P., et al.
(författare)
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Versatile gene-specific sequence tags for Arabidopsis functional genomics : Trancript profiling and reverse genetics applications
- 2004
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Ingår i: Genome Research. - : Cold Spring Harbor Laboratory. - 1088-9051 .- 1549-5469. ; 14:10B, s. 2176-2189
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Tidskriftsartikel (refereegranskat)abstract
- Microarray transcript profiling and RNA interference are two new technologies crucial for large-scale gene function studies in multicellular eukaryotes. Both rely on sequence-specific hybridization between complementary nucleic acid strands, inciting us to create a collection of gene-specific sequence tags (GSTs) representing at least 21,500 Arabidopsis genes and which are compatible with both approaches. The GSTs were carefully selected to ensure that each of them shared no significant similarity with any other region in the Arabidopsis genome. They were synthesized by PCR amplification from genomic DNA. Spotted microarrays fabricated from the GSTs show good dynamic range, specificity, and sensitivity in transcript profiling experiments. The GSTs have also been transferred to bacterial plasmid vectors via recombinational cloning protocols. These cloned GSTs constitute the ideal starting point for a variety of functional approaches, including reverse genetics. We have subcloned GSTs on a large scale into vectors designed for gene silencing in plant cells. We show that in planta expression of GST hairpin RNA results in the expected phenotypes in silenced Arabidopsis lines. These versatile GST resources provide novel and powerful tools for functional genomics.
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| 3. |
- Brooks, R. F., et al.
(författare)
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Reliable data for high-temperature viscosity and surface tension : results from a European project
- 2001
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Ingår i: High Temperatures-High Pressures. - : Old City Publishing, Inc. - 0018-1544 .- 1472-3441. ; 33:6, s. 631-637
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Tidskriftsartikel (refereegranskat)abstract
- Results are reported of work carried out for the European Commission under contract number MAT1 CT 940006, between 1994 and 1998. The project provided recommended surface tension - temperature relationships for copper and nickel samples prepared by cold crucible melting to +/-3%, by the levitation and sessile drop methods, and values were also obtained for iron and gold in preliminary studies. For copper, (gamma /N m(-1)) = 1.304 - 0.289 x 10(-3)[(theta - 1083)/degreesC]; for nickel, (gamma /N m(-1)) = 1.781 - 0.285 x 10(-3)[(theta - 1455)/degreesC]. Comparison of terrestrial measurements with those made in microgravity have shown that the Cumming's equation is a reliable and satisfactory method of correcting the surface tensions derived with the levitated drop method for the effect of electromagnetic pressure. For measurements with an oscillating cylinder viscometer, recommended viscosity-temperature relationships are given for copper, nickel, and iron. For copper, log(10)(eta /mPa s) = -0.2791 + [1245.5/(T/K)]; for nickel, log(10)(q/mPa s) = -0.5038 + [2029/(T/K)]; and for iron, log(10)(eta /mPa s) = -0.6494 + [2568/(T/K)].
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| 5. |
- Giesecke, T., et al.
(författare)
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Evidence of augmented central pain processing in idiopathic chronic low back pain
- 2004
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Ingår i: Arthritis Rheum. - : Wiley. - 0004-3591. ; 50:2, s. 613-23
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Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVE: For many individuals with chronic low back pain (CLBP), there is no identifiable cause. In other idiopathic chronic pain conditions, sensory testing and functional magnetic resonance imaging (fMRI) have identified the occurrence of generalized increased pain sensitivity, hyperalgesia, and altered brain processing, suggesting central augmentation of pain processing in such conditions. We compared the results of both of these methods as applied to patients with idiopathic CLBP (n = 11), patients with widespread pain (fibromyalgia; n = 16), and healthy control subjects (n = 11). METHODS: Patients with CLBP had low back pain persisting for at least 12 months that was unexplained by MRI/radiographic changes. Experimental pain testing was performed at a neutral site (thumbnail) to assess the pressure-pain threshold in all subjects. For fMRI studies, stimuli of equal pressure (2 kg) and of equal subjective pain intensity (slightly intense pain) were applied to this same site. RESULTS: Despite low numbers of tender points in the CLBP group, experimental pain testing revealed hyperalgesia in this group as well as in the fibromyalgia group; the pressure required to produce slightly intense pain was significantly higher in the controls (5.6 kg) than in the patients with CLBP (3.9 kg) (P = 0.03) or the patients with fibromyalgia (3.5 kg) (P = 0.006). When equal amounts of pressure were applied to the 3 groups, fMRI detected 5 common regions of neuronal activation in pain-related cortical areas in the CLBP and fibromyalgia groups (in the contralateral primary and secondary [S2] somatosensory cortices, inferior parietal lobule, cerebellum, and ipsilateral S2). This same stimulus resulted in only a single activation in controls (in the contralateral S2 somatosensory cortex). When subjects in the 3 groups received stimuli that evoked subjectively equal pain, fMRI revealed common neuronal activations in all 3 groups. CONCLUSION: At equal levels of pressure, patients with CLBP or fibromyalgia experienced significantly more pain and showed more extensive, common patterns of neuronal activation in pain-related cortical areas. When stimuli that elicited equally painful responses were applied (requiring significantly lower pressure in both patient groups as compared with the control group), neuronal activations were similar among the 3 groups. These findings are consistent with the occurrence of augmented central pain processing in patients with idiopathic CLBP.
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| 6. |
- Grant, LM, et al.
(författare)
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Influence of surface hydrophobicity on the layer properties of adsorbed nonionic surfactants
- 2000
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Ingår i: Langmuir. - 0743-7463 .- 1520-5827. ; 16, s. 2285-2291
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Tidskriftsartikel (refereegranskat)abstract
- The structure of adsorbed octa(oxyethlyene) n-dodecyl ether (C12E8) layers at a series of gold-alkanethiolate surfaces was investigated by atomic force microscopy. The hydrophobicity of the surfaces was systematically varied by changing the relative amounts of chemisorbed thiohexadecane (CH3(CH2)15SH) and thiohexadecanol (CH2OH(CH2)15SH) surface groups. This allowed complete control over the hydrophobicity of the surface. Adsorption was studied on five different thiol-modified gold surfaces prepared from solutions containing 0%, 25%, 50%, 75%, and 100% hexadecane thiol, respectively (henceforth referred to as 0% CH3, 25% CH3, 50% CH3, 75% CH3, and 100% CH3). The following general evolution of the adsorbed layer morphology with increasing surface hydrophobicity was observed: diffuse micellar coverage; dense micellar coverage; bilayer; and finally a monolayer structure at the most hydrophobic surface. The adsorbed layer structure observed at the different surfaces was interpreted in terms of the effective interaction between different parts of the surfactant and the solid surface. On the basis of our adsorption data, we also infer that hydrophobic interactions are the main driving force for adsorption of ethylene oxide segments at partially hydroxylated surfaces (such as silica) and that hydrogen bonding reduces the free energy penalty of displacing water.
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| 7. |
- Grant, LM, et al.
(författare)
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Normal and lateral forces between lipid covered solids in solution: Correlation with layer packing and structure
- 2002
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Ingår i: Biophysical Journal. - 0006-3495 .- 1542-0086. ; 82, s. 1373-1385
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Tidskriftsartikel (refereegranskat)abstract
- We report on the normal and lateral forces between controlled-density mono- and bilayers of phospholipid co-adsorbed onto hydrophobic and hydrophilic solid supports, respectively. Interactions between 1,2-dioleoyl-sn-glycero-3-phosphocholine layers were measured using an atomic force microscope. Notable features of the normal force curves (barrier heights and widths) were found to correlate with the thickness and density of the supported lipid layers. The friction and normal force curves were also found interrelated. Thus, very low friction values were measured as long as the supported layer(s) resisted the normal pressure of the tip. However, as the applied load exceeded the critical value needed for puncturing the layers, the friction jumped to values close to those recorded between bare surfaces. The lipid layers were self-healing between measurements, but a significant hysteresis was observed in the force curves measured on approach and retraction, respectively. The study shows the potential of using atomic force microscopy for lipid layer characterization both with respect to structure and interactions. It further shows the strong lubricating effect of adsorbed lipid layers and how this varies with surface density of lipids. The findings may have important implications for the issue of joint lubrication.
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| 9. |
- Philips, Matthew F., et al.
(författare)
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Neuroprotective and behavioral efficacy of nerve growth factor-transfected hippocampal progenitor cell transplants after experimental traumatic brain injury
- 2001
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Ingår i: Journal of Neurosurgery. - 0022-3085. ; 94:5, s. 765-765
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Tidskriftsartikel (refereegranskat)abstract
- OBJECT: Immortalized neural progenitor cells derived from embryonic rat hippocampus (HiB5), were transduced ex vivo with the gene for mouse nerve growth factor (NGF) to secrete NGF (NGF-HiB5) at 2 ng/hr/10(5) cells in culture. METHODS: Fifty-nine male Wistar rats weighing 300 to 370 g each were anesthetized with 60 mg/kg sodium pentobarbital and subjected to lateral fluid-percussion brain injury of moderate severity (2.3-2.4 atm, 34 rats) or sham injury (25 rats). At 24 hours postinjury, 2 microl (150,000 cells/microl) of [3H]thymidine-labeled NGF-HiB5 cells were transplanted stereotactically into three individual sites in the cerebral cortex adjacent to the injury site (14 rats). Separate groups of brain-injured rats received nontransfected (naive [n])-HiB5 cells (12 animals) or cell suspension vehicle (eight animals). One week postinjury, animals underwent neurological evaluation for motor function and cognition (Morris water maze) and were killed for histological, autoradiographic, and immunocytochemical analysis. Viable HiB5 cell grafts were identified in all animals, together with reactive microglia and macrophages located throughout the periinjured parenchyma and grafts (OX-42 immunohistochemistry). Brain-injured animals transplanted with either NGF-HiB5 or n-HiB5 cells displayed significantly improved neuromotor function (p < 0.05) and spatial learning behavior (p < 0.005) compared with brain-injured animals receiving microinjections of vehicle alone. A significant reduction in hippocampal CA3 cell death was observed in brain-injured animals receiving transplants of NGF-HiB5 cells compared with those receiving n-HiB5 cells or vehicle (p < 0.025). CONCLUSIONS: This study demonstrates that immortalized neural stem cells that have been retrovirally transduced to produce NGF can markedly improve cognitive and neuromotor function and rescue hippocampal CA3 neurons when transplanted into the injured brain during the acute posttraumatic period.
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