Impact of combinatorial dysfunctions of Tet2 and Ezh2 on the epigenome in the pathogenesis of myelodysplastic syndrome.

Somatic inactivating mutations in epigenetic regulators are frequently found in combination in myelodysplastic syndrome (MDS). However, the mechanisms by which combinatory mutations in epigenetic regulators promote the development of MDS remain unknown. Here we performed epigenomic profiling of hematopoietic progenitors in MDS mice hypomorphic for Tet2 following the loss of the polycomb-group gene Ezh2 (Tet2KD/KDEzh2Δ/Δ). Aberrant DNA methylation propagated in a sequential manner from a Tet2-insufficient state to advanced MDS with deletion of Ezh2. Hyper-differentially methylated regions (hyper-DMRs) in Tet2KD/KDEzh2Δ/Δ MDS hematopoietic stem/progenitor cells were largely distinct from those in each single mutant and correlated with transcriptional repression. Although Tet2 hypomorph was responsible for enhancer hypermethylation, the loss of Ezh2 induced hyper-DMRs that were enriched for CpG islands of polycomb targets. Notably, Ezh2 targets largely lost the H3K27me3 mark while acquiring a significantly higher level of DNA methylation than Ezh1 targets that retained the mark. These findings indicate that Ezh2 targets are the major targets of the epigenetic switch in MDS with Ezh2 insufficiency. Our results provide a detailed trail for the epigenetic drift in a well-defined MDS model and demonstrate that the combined dysfunction of epigenetic regulators cooperatively remodels the epigenome in the pathogenesis of MDS.

[Potential of photodynamic therapy with a second-generation sensitizer: mono-L-aspartyl chlorin e6].

We evaluated the potential for photodynamic therapy of a new photosensitizer, mono-L-aspartyl chlorin e6 (NPe6), whose tissue uptake and clearance are rapid. Firstly, NPe6 solution was exposed to a diode laser at 664 nm that fits the maximum absorption of NPe6. Immediately after laser exposure, the solution was subjected to electron spin resonance (ESR) to detect generation of activated oxygen species. With ESR, O2-, .OH, and 1O2 were detected in the NPe6 solution. O2- and .OH disappeared when superoxide dismutase was added to the solution. Next, the fundi of pigmented rabbits were irradiated with the diode laser after administration of NPe6. Histopathologically, there was necrosis of endothelial cells and vascular occlusion in the choroid, especially in the choriocapillaris. Retinal pigment epithelial cells were also injured. In contrast, the sensory retina was undamaged even after use of a large amount of dye. This new combination of NPe6 and diode laser at 664 nm enables us to occlude choroidal vessels efficiently by generating various activated oxygen species. Thus, photodynamic therapy with NPe6 is potentially useful in treating retinochoroidal involvement.

Structural comparison and chromosomal localization of the human and mouse IL-13 genes.

The genomic structure of the recently described cytokine IL-13 has been determined for both human and mouse genes. The nucleotide sequence of a 4.6-kb DNA segment of the human gene is described. The human IL-13 gene (IL13) occurs as a single copy in the haploid genome and maps to human chromosome 5. A 4.3-kb DNA fragment of the mouse IL-13 gene (IL13) has been sequenced and found to occur as a single copy, mapping to mouse chromosome 11. Intrachromosomal mapping studies revealed that both genes contain four exons and three introns and show a high degree of sequence identity throughout their length. Potential recognition sequences for transcription factors that are present in the 5'-flanking region and are conserved between both genes include IFN-responsive elements, binding sites for AP-1, AP-2 and AP-3, and NF-IL 6 site, and a TATA-like sequence. Both genes map to chromosomal locations adjacent to genes encoding other cytokines, including IL-3, GM-CSF, IL-5, and IL-4, suggesting that IL-13 is another member of this cytokine gene family that may have arisen by gene duplication.

Iatrogenic acute spinal epidural abscess with septic meningitis: MR findings.

A contaminated catheter used in epidural anesthesia in a 71-year-old female produced acute epidural abscess and septic meningitis. Methicillin-resistant Staphylococcus aureus (MRSA) was detected in a culture of the epidural pus. Both T1- and T2-weighted MR images showed low intensity mass lesion compressing the thecal sac behind the vertebral body L3. The low intensity lesion was probably pus with gas component. In these low intensity lesions in MR findings with gas component, MR was superior to myelography because it visualized both the degree of compression to the thecal sac and extension of the lesion in all directions.

[Monitoring of the spinal cord function using evoked spinal cord potentials].

Monitoring of the spinal cord function by evoked spinal cord potentials (ESP) has come to be used widely with the establishment of a safe technique using an epidural electrode and the great improvement in medical electronic devices. According to other investigations ESP has been recorded through an electrode placed in the posterior epidural space, it is well known that residual ESP can be actually recorded after transection of the posterior half of the spinal cord and that ESP shows no change after a small transection of the anterior column. If ESP originating from the anterior half of the spinal cord is recorded, it is very useful for monitoring during anterior decompression surgery for myelopathy such as ossification of the posterior longitudinal ligament and cervical spondylosis. The purpose of this report is to analyse ESP originating from the anterior half of the spinal cord experimentally and further investigate the ESP monitoring clinically. Sixty cats were used in this study. ESP recording principally consisted of stimulation to the spinal cord at the level of thoracic or lumbar region and recording of ESP with a monopolar electrode placed in the cervical epidural space. Changes in the ESP after transection and compression of the spinal cord were analysed. Anterior vertebrectomy and laminectomy at C2 and C6 were performed for the purpose of placing an electrode correctly and facilitating transection or compression of the spinal cord. Recording electrodes were placed on both the anterior and posterior surfaces of the dura mater at C2. As the result of this study, it is apparent that there exists the ESP originating from the anterior half of the spinal cord. Such ESPs are more accurately recorded through an anterior placed electrode rather than through that placed posteriorly.

Black lipid membranes as a model for intestinal absorption of drugs.

Black lipid membranes were generated in isotonic buffer (pH 4-5 and pH 6-5) from egg phosphatidylcholine and intestinal lipid, and the permeability to salicylamide, salicylic acid, p-aminobenzoic acid and tryptophan of these membranes was studied. Electrical resistance of intestinal lipid membranes was higher than that of phosphatidylcholine membranes. The presence of cholesterol produced an increase in the electrical resistance of black lipid membranes and a small decrease in the permeability of membranes to drugs. The permeability coefficient of salicylamide, an uncharged drug, was much larger than the coefficients of the charged drugs examined. The values for salicylic acid and p-aminobenzoic acid were much larger than comparable values predicted from their partition coefficients. Intestinal lipid membranes were more permeable to acidic drugs than phosphatidylcholine membranes. It is suggested that phospholipids and other lipid components of the small intestine may play an important role in the membrane permeability to acidic drugs. This method may be of interest in studying the complex processes of drug absorption from intestine.