Reduction of metal artefacts in musculoskeletal MR imaging.

The purpose of this article is to present a educational overview of practical tips to deal with metal artefacts in clinical musculoskeletal MRI. A brief theoretical explanation to understand the cause of metal artefacts is provided followed by a discussion on parameters to reduce these metal artefacts. Effects of adjustable parameters are demonstrated both in a volunteer with a titanium screw and a saline bag attached to the shoulder and in a in vitro experiment. These parameters include positioning of the patient with the long axis of metallic hardware parallel to B0, use of fast spin echo sequences, use of inversion recovery fat suppression, swapping phase and frequency encoding direction, use of view angle tilting, increasing the read-out bandwidth, and decreasing voxel size.

Hypoxic repression of lactate dehydrogenase-B in retina.

In order to elucidate mechanisms controlling hypoxia induced gene repression in the retina, we studied expression of the lactate dehydrogenase gene LDH-B. RT/PCR was used to isolate cDNA fragments of the LDH-A and LDH-B genes from both rat and chick. Northern analysis was used to measure LDH-B mRNA expression and in situ hybridization to localize LDH-B mRNA during development of the avascular chick retina. Primary retinal cultures of rat and chick were subjected to hypoxia and reperfusion paradigms, and levels of LDH-A and LDH-B mRNA expression were measured by Northern and semi-quantitative RT/PCR analyses. Northern analysis demonstrated that LDH-B mRNA is developmentally regulated in the chick retina according to a time course that correlates with oxygen availability. In developing chick retina in situ hybridization localized LDH-B mRNA to aerobic regions of the retina, coincident with the distribution of active mitochondria. Northern and semi-quantitative RT/PCR analyses demonstrated that LDH-B mRNA levels decrease in primary rat and chick retinal cells in response to hypoxia. LDH-B mRNA increased to control levels in retinal cells exposed to hypoxia then reperfused with oxygen, while the opposite was true for LDH-A, an hypoxia inducible gene. These data demonstrate that LDH-B gene repression after hypoxia and reactivation after oxygen reperfusion occurs in both vascular and avascular retinal cells. Oxygen regulated expression of LDH-B is opposite and complementary to that of LDH-A and provides a system to elucidate the mechanisms underlying hypoxic gene repression and reactivation after reperfusion. Mechanisms controlling gene repression and reactivation based on oxygen availability are likely to be critically involved in ischemic damage and neovascular retinopathy.

The 5' flanking regions of IRBP and arrestin have promoter activity in primary embryonic chicken retina cell cultures.

Primary cultures of embryonic chicken cells from various tissues were transiently transfected with plasmid vectors containing reporter genes linked to a 1.8 kb fragment of the mouse interphotoreceptor retinoid-binding protein (IRBP) 5' flanking region, a 1.5 kb fragment of the mouse arrestin 5' flanking region, or a 3.4 kb sequence of the bovine arrestin 5' flanking region. Promoter activity was evident in retina-derived cells, but not in fibroblasts or cells from whole brain. Transfection response also varied with transfection method, plasmid DNA concentration, post-transfection incubation time, and cell density. The data suggest that the primary embryonic chicken retinal cell culture system is a useful tool in studying photoreceptor-specific gene regulation.

Synergistic transcriptional activation of the IL-8 gene by NF-kappa B p65 (RelA) and NF-IL-6.

Transcriptional activation of the IL-8 gene by several inflammatory mediators, including the cytokines IL-1 and TNF-alpha, is mediated through sequences located between nucleotide -94 and -71 of the IL-8 promoter. Because adjacent binding sites for the inducible transcription factors NF-kappa B and NF-IL-6 are located within this region, we examined the functional interaction of these two transcription factor families in IL-8 gene regulation. Maximal transcriptional activation by PMA in Jurkat T lymphocytes was shown to require intact binding sites for both NF-kappa B and NF-IL-6. Electrophoretic mobility shift analysis indicates that NF-IL-6, as well as other related members of this family, bind specifically to the NF-IL-6 site in the IL-8 promoter. In addition, NF-kappa B p65 (RelA), but not NF-kappa B p50 (NFKB1), binds specifically to the NF-kappa B site. When incubated together, RelA and NF-IL-6/C/EBP form a ternary complex with this region of the IL-8 promoter; this binding is dependent on intact binding sites for both NF-IL-6 and RelA. Transient cotransfection analyses indicate that the cooperative association of NF-IL-6 and RelA with the IL-8 promoter results in synergistic transcriptional activation. Mutational analyses of RelA demonstrate that the C-terminal transactivation domain and the DNA binding domain are required for synergistic activation with NF-IL-6. In addition, overexpression of the NF-kappa B inhibitor molecule, I kappa B, abolished the RelA- and RelA/NF-IL-6-dependent synergistic activation. These data demonstrate that RelA and members of the C/EBP/NF-IL-6 family can functionally cooperate in transcriptional activation of the IL-8 gene and suggest a common mechanism for inducible regulation of cytokine gene expression.

Requirement for nuclear factor (NF)-kappa B p65 and NF-interleukin-6 binding elements in the tumor necrosis factor response region of the granulocyte colony-stimulating factor promoter.

Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic growth factor produced by mesenchymal and myeloid cells following activation by inflammatory stimuli. It has previously been shown that a region of the G-CSF promoter, (-200 to -165) containing the decanucleotide CK-1 element and two repeated sequences that resemble nuclear factor (NF)-interleukin-6 (IL-6) binding sites, is required for activation of the G-CSF gene by tumor necrosis factor-alpha (TNF-alpha) and IL-1 beta. We now show that the NF-kappa B p65 protein can bind to and activate this TNF response region. There are several unusual features of this p65 interaction with the TNF response region. First, NF-kappa B p65 but not the related NF-kappa B p50 binds to the CK-1 element and a p50/65 hybrid protein that relies on the p50 rel homology domain for DNA binding does not transactivate the TNF response region. Second, p65 transactivation of this region is cell specific and requires not only its own binding site but also the NF-IL6 consensus sites. NF-IL6 also binds to the TNF response region of the G-CSF promoter. Electrophoretic mobility shift studies show that p65 and NF-IL6 can bind cooperatively to the TNF response region. The ability of this region to respond to TNF-alpha or p65 is correlated with the ability to form the p65/NF-IL6 ternary complex.

HTLV-1 tax activation of the GM-CSF and G-CSF promoters requires the interaction of NF-kB with other transcription factor families.

The trans-activator protein, tax, from the human T leukemia virus type 1 (HTLV-1) trans-activates both viral and cellular genes. It has previously been shown that granulocyte macrophage-colony stimulating factor (GM-CSF) is constitutively expressed in HTLV-1 infected cells and in cells artificially expressing tax. We show here that the GM-CSF promoter is tax responsive in fibroblasts and T cells, whereas the granulocyte (G)-CSF promoter is tax responsive only in fibroblasts. The tax protein can activate cellular genes through a least two families of transcription factors; the NF-kB/rel and CREB/ATF families. We have used mutant tax proteins to show that the activation of NF-kB proteins is essential for tax trans-activation of both the GM-CSF and G-CSF promoters. The ability of tax to activate CREB/ATF proteins is also essential for GM-CSF transactivation. We have identified a 44 bp region of the GM-CSF promoter that contains tax responsive elements. This region contains a classical NF-kB site, a CK-1 element that can bind the NF-kB p65 protein, as well as a putative ATF binding site. The tax response of the G-CSF promoter requires not only the conserved CK-1 sequence but also an adjacent NF-IL6 binding site that may explain the cell restricted function of the G-CSF promoter.