Kawasaki disease (KD) is an acute vasculitis of childhood that predominantly affects the coronary arteries. We investigated single nucleotide polymorphisms (SNPs) of the tryptophan hydroxylase 2 (TPH2) gene as risk factors for KD with coronary artery lesions (CALs) in Korean children. We genotyped two SNPs [rs7305115 (exon 7) and rs4290270 (exon 9)] using direct sequencing in 101 KD and 256 control subjects. To analyze the genetic data, SNPStats, SNPAnalyzer, and Helixtree programs were used. The genotype analysis of rs7305115 and rs4290270 showed no significant differences between KD and control groups. However, we found a statistically significant association between the two SNPs and the development of CALs in KD (p < 0.05). The minor homozygous genotype (rs7305115, AA genotype and rs42901270, AA genotype) of each SNP showed increased susceptibility to risk of CALs in KD patients. These results suggest that TPH2 may be associated with the development of KD with CALs in Korean children.

OBJECTIVES: OK-432 has been widely used to treat lymphangioma and ranula; however, there are few studies for its use in treatment of branchial cleft cyst (BCC). We conducted this study to evaluate the effectiveness of sclerotherapy using OK-432 in treatment of BCC. STUDY DESIGN AND SETTING: Case series with planned data collection. SUBJECTS AND METHODS: From 2004 to 2007, we treated 23 patients with BCC using OK-432 sclerotherapy. Of these 23 patients, 18 had unilocular cysts and five had multilocular cysts. The sizes of the BCCs were measured and compared before and after treatment. RESULTS: Of the 23 cases, 14 (60.8%) showed complete regression; all of these were unilocular cysts. Of the remaining individuals with unilocular cysts, only one patient failed to show any response. This individual subsequently underwent surgical excision. A total of five patients with multilocular cysts showed no or partial response and subsequently underwent surgical excision. Minor adverse effects including fever and local pain were reported by 13 (56.5%) patients. CONCLUSION: These results suggest that sclerotherapy using OK-432 is an effective and safe treatment modality for BCC, especially for unilocular cysts. Sclerosing of unilocular BCC with OK-432 should therefore be considered before surgical excision.

Coronary artery disease (CAD) is caused by a thrombotic occlusion or spasm of the coronary artery. Association of genetic variants with susceptibility to CAD has been reported in various populations, but the association should be replicated in other populations to establish the role of genetic variants in CAD. We conducted a case-control study with a total of 1480 CAD cases and 2115 controls from two East Asian populations, Japanese and Korean, to validate the associations of CAD with eight single nucleotide polymorphisms (SNPs) in eight loci, which were identified from large-scale whole-genome association studies in Europeans or East Asians. Among the tested SNPs, one SNP in BRAP (rs11066001) showed a significant association in allele frequency distribution with CAD in both the Japanese (Odds ratio (OR)=1.63, 95% confidence interval (CI); 1.41-1.89, P=5.0 x 10(-11), corrected P (Pc)=4.0 x 10(-10)) and Korean populations (OR=1.68, 95% CI; 1.41-2.00, P=6.5 x 10(-9), Pc=5.2 x 10(-9)), and a meta-analysis showed a significant association in the East Asian populations (OR=1.65, 95% CI; 1.48-1.85, P=1.8 x 10(-18), Pc=1.4 x 10(-17)), whereas no evidence of association was found for the other SNPs. In addition, a combined analysis of BRAP and another CAD locus on 9p21 suggested that these loci had a synergistic role in the susceptibility. Failure to replicate the association with the other SNPs, which were reported in the European populations, suggested that their contributions to CAD were not large enough to be readily captured in the East Asian populations.Journal of Human Genetics advance online publication, 28 August 2009; doi:10.1038/jhg.2009.87.

Fired cartridge cases are a common type of evidence found at crime scenes. However, due to the high chamber temperatures and touch nature of this evidence, DNA testing is not commonly sought because it is believed DNA is only present in low levels, whether it is due to initial low levels of DNA and/or DNA degradation from the heat or inhibition of the PCR reaction. Moreover, very few laboratories report STR typing success with fired cases. This study focused on obtaining STR profiles from fired cartridge cases using the AmpFlSTR((R)) MiniFiler kit, which is designed to amplify DNA from low level, inhibited, and degraded samples. Comparisons to other STR amplification kits were also conducted. In attempt to simulate casework, random individuals loaded cartridges into a firearm. DNA was recovered from the fired cartridge cases using the double swab technique and extracted using an automated large volume DNA IQ method. Initially, testing focused on known shedders handling cartridges for 30s prior to firing. A significantly greater number of alleles was obtained following amplification with the MiniFiler kit versus the PowerPlex((R)) 16 BIO kit. No alleles were observed using the Identifiler((R)) kit. In an attempt to better simulate casework, a random selection of laboratory personnel handled shotshells for as long as needed to load and fire the weapon. In this mock sample study, the MiniFiler kit successfully amplified an average of 22% of expected alleles from DNA recovered from shotshell cases versus the PowerPlex((R)) 16 BIO kit where an average of 7% of alleles were observed. However, the total number of alleles obtained from the two kits was not significantly different. The quality of the DNA obtained from fired cases was studied with evidence of inhibition in at least 11% of shotshell case samples. After swabbing the head and the hull of three shotshell cases separately, a significantly greater number of alleles was obtained from the hull as opposed to the head of the fired shotshell case. In addition, after firing, various internal firearm surfaces were swabbed, including the chamber of barrel, ejection port, and breechface, in an attempt to obtain amplifiable DNA. DNA was obtained from the chamber of the barrel and was amplifiable using the MiniFiler kit, although mixtures were obtained with extensive drop-in and drop-out making this analysis unlikely to aid an investigation.

OBJECTIVES: Canalith repositioning procedure (CRP) provides rapid and long-lasting relief of symptoms in most patients with benign paroxysmal positional vertigo. However, some patients express nonspecific symptoms such as anxiety or discomfort after treatment, even after the disappearance of nystagmus and vertigo. The purpose of this study was to assess the residual symptoms after CRP in patients with benign paroxysmal positional vertigo using the Dizziness Handicap Inventory (DHI) in a questionnaire format. STUDY DESIGN AND SETTING: Controlled, prospective study. SUBJECTS AND METHODS: CRP was performed in 135 patients until nystagmus and vertigo disappeared. Patients were asked to complete the questionnaire before and 5 to 7 days after treatment. A control group of 135 normal volunteers was selected and cross-matched according to the age and sex of the patient group. The data were compared for the pre-CRP, post-CRP, and control groups. RESULTS: There was a significant improvement in the DHI scores when comparing the pre- and post-CRP groups (P = 0.000), although six items showed incomplete improvement. Subsequent comparison of DHI scores between the control group and the post-CRP group still showed a difference in some items so that the improvement was incomplete. CONCLUSION: Even after successful CRPs, Dizziness Handicap Inventory scores indicated that residual subjective symptoms may remain. Thus, additional follow-up and management are important for these patients.

Coronary artery disease (CAD) is based on the atherosclerosis of coronary artery and may manifest with myocardial infarction or angina pectoris. Although it is widely accepted that genetic factors are linked to CAD and several disease-related genes have been reported, only a few could be replicated suggesting that there might be some other CAD-related genes. To identify novel susceptibility loci for CAD, we used microsatellite markers in the screening and found six different candidate CAD loci. Subsequent single nucleotide polymorphism (SNP) association studies revealed an association between CAD and megakaryoblastic leukemia factor-1 gene (MKL1). The association with a promoter SNP of MKL1,-184C > T, was found in a Japanese population and the association was replicated in another Japanese population and a Korean population. Functional analysis of the MKL1 promoter SNP suggested that the higher MKL1 expression was associated with CAD. These findings suggest that MKL1 is involved in the pathogenesis of CAD.

BACKGROUND: EWS-FLI1 is a chimeric ETS transcription factor that is, due to a chromosomal rearrangement, specifically expressed in Ewing's sarcoma family tumors (ESFT) and is thought to initiate the development of the disease. Previous genomic profiling experiments have identified EWS-FLI1-regulated genes and genes that discriminate ESFT from other sarcomas, but so far a comprehensive analysis of EWS-FLI1-dependent molecular functions characterizing this aggressive cancer is lacking. METHODOLOGY/PRINCIPAL FINDINGS: In this study, a molecular function map of ESFT was constructed based on an integrative analysis of gene expression profiling experiments following EWS-FLI1 knockdown in a panel of five ESFT cell lines, and on gene expression data from the same platform of 59 primary ESFT. Out of 80 normal tissues tested, mesenchymal progenitor cells (MPC) were found to fit the hypothesis that EWS-FLI1 is the driving transcriptional force in ESFT best and were therefore used as the reference tissue for the construction of the molecular function map. The interrelations of molecular pathways were visualized by measuring the similarity among annotated gene functions by gene sharing. The molecular function map highlighted distinct clusters of activities for EWS-FLI1 regulated genes in ESFT and revealed a striking difference between EWS-FLI1 up- and down-regulated genes: EWS-FLI1 induced genes mainly belong to cell cycle regulation, proliferation, and response to DNA damage, while repressed genes were associated with differentiation and cell communication. CONCLUSIONS/SIGNIFICANCE: This study revealed that EWS-FLI1 combines by distinct molecular mechanisms two important functions of cellular transformation in one protein, growth promotion and differentiation blockage. By taking MPC as a reference tissue, a significant EWS-FLI1 signature was discovered in ESFT that only partially overlapped with previously published EWS-FLI1-dependent gene expression patterns, identifying a series of novel targets for the chimeric protein in ESFT. Our results may guide target selection for future ESFT specific therapies.

Coronary artery disease (CAD) has become a major health problem in many countries because of its increasing prevalence and high mortality. Recently, an association of a functional sequence variation,-8C>G, in the human proteasome subunit alpha type 6 gene (PSMA6) with the susceptibility to CAD was reported. To validate the association, we investigated a total of 1330 cases and 2554 controls from Japanese and Korean populations for PSMA6 genotypes, and no evidence of the association was obtained in both Japanese (odds ratio (OR)=1.03, 95% confidence interval (CI); 0.90-1.19, P=0.66, allele count model) and Korean populations (OR=1.00, 95% CI; 0.86-1.17, P=0.95, allele count model). However, when a meta-analysis of data from this study and previously reported six replication studies was done, OR was 1.08 for the G allele (95% CI; 1.02-1.14, P=0.0057), suggesting that the contribution of PSMA6 to CAD was not large enough to be readily replicated. Further studies are required to establish the contribution of this variant in the susceptibility to CAD.Journal of Human Genetics advance online publication, 13 March 2009; doi:10.1038/jhg.2009.22.

Kawasaki disease (KD) is an acute febrile vasculitis that predominantly affects infants and young children. Tissue inhibitors of matrix metalloproteinases (TIMPs) comprise a family of four members, of which TIMP4 is characterized by its restriction to cardiovascular structures. In KD pathophysiology, TIMP4 is considered to be involved in the development of coronary artery lesions (CALs). Therefore, this study investigated single-nucleotide polymorphisms (SNPs) of the TIMP4 gene as risk factors for KD with CALs in Korean children. To observe this association, two SNPs (rs3755724,-55C/T, promoter; rs17035945, 3'-untranslated region) were genotyped in TIMP4 using direct sequencing. There were no SNPs in the coding region of TIMP4, and two SNPs were selected in the exon and promoter regions. This study recruited 250 control and 101 KD subjects. For data analysis, SNPStats, SNPAnalyzer, and Helixtree programs were used. These SNPs were not associated with KD. However, in the recessive model, a significant association was found between rs3755724 and the development of CALs in KD (P = 0.02; odds ratio, 0.31; 95% confidence interval, 0.11-0.85). The minor allele (C) of rs3755724 showed the susceptibility of CALs to risk in KD patients. These results suggest that TIMP4 is related to the development of KD with CALs in Korean children.

Atomic force microscopy (AFM) provides the possibility to map the 3D structure of viewed objects with a nanometric resolution, which cannot be achieved with other imaging methods such as conventional video imaging and confocal fluorescent microscopy. Video imaging with CCD cameras can provide an analysis of biological events with a temporal and spatial resolution not possible with AFM, while confocal imaging allows the simultaneous acquisition of immunofluorescence images. In this communication we present a simple method to combine AFM and confocal images to study differentiating embryonic stem (ES) cells-derived and dorsal root ganglia (DRG) neurons in culture. Neurons were grown on coverslips with micrometric markers that allow finding and imaging the same neuron with different microscopes. AFM and confocal images were registered using conventional methods used in Computer Science. The combination of these two techniques allows relating functional properties to morphological features of imaged neurons.