We present the first Korean individual genome sequence (SJK) and analysis results. The diploid genome of a Korean male was sequenced to 28.95-fold redundancy using the Illumina paired-end sequencing method. SJK covered 99.9% of the NCBI human reference genome. We identified 420,083 novel single nucleotide polymorphisms (SNPs) that are not in the dbSNP database. Despite a close similarity, significant differences were observed between the Chinese genome (YH), the only other Asian genome available, and SJK: (1) 39.87% (1,371,239 out of 3,439,107) SNPs were SJK-specific (49.51% against Venter's, 46.94% against Watson's, and 44.17% against the Yoruba genomes); (2) 99.5% (22,495 out of 22,605) of short indels (< 4 bp) discovered on the same loci had the same size and type as YH; and (3) 11.3% (331 out of 2920) deletion structural variants were SJK-specific. Even after attempting to map unmapped reads of SJK to unanchored NCBI scaffolds, HGSV, and available personal genomes, there were still 5.77% SJK reads that could not be mapped. All these findings indicate that the overall genetic differences among individuals from closely related ethnic groups may be significant. Hence, constructing reference genomes for minor socio-ethnic groups will be useful for massive individual genome sequencing.

The corpus callosum (CC) is the major conduit for information transfer between the cerebral hemispheres and plays an integral role in relaying sensory, motor and cognitive information between homologous cortical regions. The majority of fibers that make up the CC arise from large pyramidal neurons in layers III and V, which project contra-laterally. These neurons degenerate in Huntington’s disease (HD) in a topographically and temporally selective way. Since any focus of cortical degeneration could be expected to secondarily de-afferent homologous regions of cortex, we hypothesized that regionally selective cortical degeneration would be reflected in regionally selective degeneration of the CC. We used conventional T1-weighted, diffusion tensor imaging (DTI), and a modified corpus callosum segmentation scheme to examine the CC in healthy controls, huntingtin gene-carriers and symptomatic HD subjects. We measured mid-sagittal callosal cross-sectional thickness and several DTI parameters, including fractional anisotropy (FA), which reflects the degree of white matter organization, radial diffusivity, a suggested index of myelin integrity, and axial diffusivity, a suggested index of axonal damage of the CC. We found a topologically selective pattern of alterations in these measures in pre-manifest subjects that were more extensive in early symptomatic HD subjects and that correlated with performance on distinct cognitive measures, suggesting an important role of for disrupted inter-hemispheric transfer in the clinical symptoms of HD. Our findings provide evidence for early degeneration of commissural pyramidal neurons in the neocortex, loss of cortico-cortical connectivity, and functional compromise of associative cortical processing.

Background: Metformin exhibits anti-inflammatory effects. Results: In murine macrophages, metformin induces activating transcription factor-3 (ATF-3) in parallel with protective effects against LPS-induced inflammation. Conclusion: Anti-inflammatory action of metformin is at least partly mediated via ATF-3 induction. Significance: This finding provides a new perspective on metformin action and novel therapeutic means of treating inflammation-related diseases, i.e. ATF-3 modulation.

The apoptotic actions of p53 require its phosphorylation by a family of phosphoinositide-3-kinase-related-kinases (PIKKs), which include DNA-PKcs and ATM. These kinases are stabilized by the TTT (Tel2, Tti1, Tti2) co-chaperone family, whose actions are mediated by CK2 phosphorylation. The inositol pyrophosphates, such as 5-diphosphoinositol pentakisphosphate (IP7), are generated by a family of inositol hexakisphosphate kinases (IP6Ks) of which IP6K2 has been implicated in p53-associated cell death. In the present study we report a novel apoptotic signaling cascade linking CK2, TTT, the PIKKs, and p53. We demonstrate that IP7, formed by IP6K2, binds CK2 to enhance its phosphorylation of the TTT complex thereby stabilizing DNA-PKcs and ATM. This process stimulates p53 phosphorylation at serine-15 to activate the cell death program in human cancer cells and in murine B cells.

Early onset torsion dystonia, the most common form of hereditary primary dystonia, is caused by a mutation in the TOR1A gene, which codes for the protein torsinA. This form of dystonia is referred to as DYT1. We have used a transgenic mouse model of DYT1 dystonia [human mutant-type (hMT)1 mice] to examine the effect of the mutant human torsinA protein on striatal dopaminergic function. Analysis of striatal tissue dopamine (DA) and metabolites using HPLC revealed no difference between hMT1 mice and their non-transgenic littermates. Pre-synaptic DA transporters were studied using in vitro autoradiography with receptors. There were again no differences in the density of striatal binding sites for these ligands. Using in vivo microdialysis in awake animals, we studied basal as well as amphetamine-stimulated striatal extracellular DA levels. Basal extracellular DA levels were similar, but the response to amphetamine was markedly attenuated in the hMT1 mice compared with their non-transgenic littermates (253 ± 71% vs. 561 ± 132%, p < 0.05, two-way ANOVA). These observations suggest that the mutation in the torsinA protein responsible for DYT1 dystonia may interfere with transport or release of DA, but does not alter pre-synaptic transporters or postsynaptic DA receptors. The defect in DA release as observed may contribute to the abnormalities in motor learning as previously documented in this transgenic mouse model, and may contribute to the clinical symptoms of the human disorder.

Figure 6. RORα activates Alox12-dependent MaR1 synthesis. (A) Seven-week-old C57BL/6 mice were fed with either LFD or HFD for 12 weeks (n = 4) or fed with MCS or MCD for 4 weeks (n = 5) (first and second panels). The LFD-fed C57BL/6 mice were treated with 5 mg/kg BW SR1078 for 5 days (n = 5) (third panel). Seven-week-old LFD-fed floxed and RORα-MKO mice were sacrificed (n = 11) (fourth panel). (B) Liver samples were obtained from the floxed and RORα-MKO mice those described in Supplemental Figure 1 (n = 5). Levels of MaR1 and RvD1 in liver tissues were measured. *P < 0.05 and **P < 0.01; ## P < 0.01 for A and B. (C) DHA-treated peritoneal macrophages (PM) and Raw 264.7 cells were treated with 5 μM SR1078 for 24 hours, or the cells were infected by lenti-shGFP or lenti-shRORα for 48 hours. Intracellular amount of MaR1 were measured. *P < 0.05 (n = 3). (D) A scheme for biosynthesis of MaR1 by LOX family. (E) Expression levels of 12-LOX protein (Alox12 mRNA) and 12/15-LOX protein (Alox15 mRNA) in liver macrophages (LM), PM, Raw 264.7, bone marrow-derived macrophages (BMDM), and hepatocytes were measured by Western blotting and qRT-PCR. (F) mRNA levels of Alox genes in the isolated LMs from floxed and RORα-MKO mice as shown in A were measured by qRT-PCR. (G) LMs were treated with SR1078 or MaR1 (left). LMs were infected by AAV-GFP/AAV-RORα or lenti-shGFP/lenti-shRORα as indicated (right). The mRNA levels of Alox12 were measured by qRT-PCR. *P < 0.05 (n = 3) for F and G. (H) Schematic representation of the mouse Alox12 promoter with the putative ROREs shown as red boxes (top). Raw 264.7 cells were transfected with the deleted Alox12 promoter-Luc reporter with empty vector (EV) or Myc-RORα. Luciferase activity was measured and normalized by β-galactosidase activity. *P < 0.05 (n = 3) (middle). Raw 264.7 cells were transfected with Myc-RORα, or cells were treated with SR1078 or MaR1. DNA fragments that contain flanking region of the ROREs on the Alox12 promoter were immunoprecipitated with indicated antibodies and then amplified by PCR (bottom). (I) DHA-treated PMs were treated with 5 μM SR1078, 5 μM baicalein, or 10 μM NCTT-956. Intracellular MaR1 content was measured. (J) LMs were treated with baicalein, or NCTT-956 in the presence or absence of DHA. The mRNA levels of Rora were measured by qRT-PCR (left). The CD206 + /CD80 + ratio of F4/80 + cells was determined by flow cytometry (right). *P < 0.05 and # P < 0.05 (n = 3) for I and J. The data represent mean ± SD. Data were analyzed by Mann-Whitney U test for simple comparisons or Kruskal-Wallis test for multiple groups.

We identified the peroxisomal proliferator response element (PPRE) in the +68/+89 region of the rat GLUT2 gene. To identify whether the putative PPRE in the GLUT2 gene (GLUT2-PPRE) is functional, GLUT2 promoter-luciferase reporter constructs were transfected into CV-1 cells. Promoter activities were increased by coexpression of peroxisomal proliferator-activated receptor (PPAR)-␥, retinoid X receptor (RXR)-␣, and treatment of their ligands; troglitazone and 9-cis retinoic acid potentiated the transactivational effects. Introduction of mutations in GLUT2-PPRE resulted in loss of transactivational effects of the PPAR-␥/RXR-␣ heterodimer. Electrophoretic mobility shift assay using nuclear extracts of CV-1 cells, which were transfected with various combinations of PPARs or RXR-␣ expression plasmids, revealed that heterodimers of PPAR-␥ and RXR-␣ preferentially bound to GLUT2-PPRE. In HIT-T15 cells, promoter activity of the rat GLUT2 gene was increased by troglitazone and 9-cis retinoic acid, and mutations of GLUT2-PPRE resulted in reduction of promoter activity. In addition, we observed increased GLUT2 transcription by troglitazone and 9-cis retinoic acid in isolated rat primary islets. These results suggested that the GLUT2-PPRE is functional and plays a significant role in gene expression of GLUT2 in pancreatic ␤-cells. This is the first report identifying PPRE in a gene involved in glucose homeostasis, linking the effect of troglitazone on the regulation of insulin secretion. Diabetes 49:1517-1524, 2000

Liver glucokinase (LGK) plays an essential role in controlling blood glucose levels and maintaining cellular metabolic functions. Expression of LGK is induced mainly regulated by insulin through sterol regulatory element-binding protein-1c (SREBP1c) as a mediator. Since LGK expression is known to be decreased in the liver of liver X receptor (LXR) knockout mice, we have investigated whether LGK might be directly activated by LXR␣. Furthermore, we have studied interrelationship between transcription factors that control gene expression of LGK. In the current studies, we demonstrated that LXR␣ increased LGK expression in primary hepatocytes and that there is a functional LXR response element in the LGK gene promoter as shown by electrophoretic mobility shift and chromatin precipitation assay. In addition, our studies demonstrate that LXR␣ and insulin activation of the LGK gene promoter occurs through a multifaceted indirect mechanism. LXR␣ increases SREBP-1c expression and then insulin stimulates the processing of the membrane-bound precursor SREBP-1c protein, and it activates LGK expression through SREBP sites in its promoter. LXR␣ also activates the LGK promoter by increasing the transcriptional activity and induction of peroxisome proliferator-activated receptor (PPAR)-␥, which also stimulates LGK expression through a peroxisome proliferator-responsive element. This activation is tempered through a negative mechanism, where a small heterodimer partner (SHP) decreases LGK gene expression by inhibiting the transcriptional activity of LXR␣ and PPAR␥ by directly interacting with their common heterodimer partner RXR␣. From these data, we propose a mechanism for LXR␣ in controlling the gene expression of LGK that involves activation through SREBP-1c and PPAR␥ and inhibition through SHP.