Comparability and utility of body composition measurement vs. anthropometric measurement for assessing obesity related health risks in Korean men.

BACKGROUND: Obesity is commonly assessed by body mass index (BMI) of which limitations come from an inability to distinguish body fat mass from lean mass. Several anthropometric measurements, including BMI, waist circumference, waist-to-height ratio and waist-to-hip ratio have been used to predict metabolic syndrome. The purpose of this study was to evaluate the utility of FMI or BF% combined with previous known anthropometric indices to assess the risk of metabolic syndrome in clinical practice. METHODS: In 5534 men visiting a hospital for health check-ups, blood tests, anthropometric measurements and body composition analysis using BIA were performed. Logistic regression analysis was performed to compare the odds ratios for metabolic syndrome and each component of metabolic syndrome among BMI, waist-to-height ratio, waist-to-hip ratio, FMI and BF%. The area under the curve (AUC) of the receiver operating characteristic curve (ROC) for metabolic syndrome was compared between several measurements. The net reclassification improvement with integrated discrimination improvement was used for assessing value of body composition measurement. RESULTS: The adjusted odds ratios of metabolic syndrome was 1.80 (95% CI, 1.71-1.89) for FMI and 1.15 (95% CI, 1.13-1.17) for BF%. Odds ratio of each metabolic component was highest for FMI among several anthropometric and body composition measurements. AUCs using the ROC curve for metabolic syndrome was highest for waist-to-height ratio, 0.823 (95% CI, 0.808-0.837) by National Cholesterol Education Program criteria. FMI caused a mild increase in integrated discrimination improvement when combined with waist-to-height ratio. CONCLUSIONS: Waist-to-height ratio seems to be the best screening tool for evaluating metabolic syndrome in Korean men, and adding FMI could result in a modest increase in integrated discrimination improvement.

Down-regulation of ATBF1 is a major inactivating mechanism in hepatocellular carcinoma.

AIMS: alpha-Fetoprotein (AFP) is frequently detected in hepatocellular carcinomas (HCCs) and AT motif binding factor 1 (ATBF1) down-regulates AFP gene expression in hepatic cells. The ATBF1 gene also inhibits cell growth and differentiation, and altered gene expression is associated with malignant transformation. The aim was to investigate the potential role of the ATBF1 gene in HCCs. METHODS AND RESULTS: Somatic mutations, allelic loss and hypermethylation of the ATBF1 gene were analysed in 76 sporadic HCCs. The level of ATBF-1 mRNA expression was analysed using quantitative real-time reverse transcriptase-polymerase chain reaction. Genetic studies of the ATBF1 gene revealed absence of somatic mutation in the hotspot region and 15 (25%) of 60 informative cases showed allelic loss at the ATBF1 locus. Hypermethylation in the intron 1 region of the ATBF1 gene was detected in only one case. Interestingly, ATBF1 mRNA expression in HCCs was significantly reduced in 55 (72.4%) samples compared with the corresponding surrounding liver tissues. Reduced expression was not statistically associated with clinicopathological parameters including stage, histological grade, infective virus type, and serum alpha-fetoprotein level. CONCLUSIONS: The ATBF1 gene may contribute to the development of HCCs via transcriptional down-regulation of mRNA expression, but not by genetic or epigenetic alterations.

KLF6 IVS1 -27G/A polymorphism with susceptibility to gastric cancers in Korean.

KLF6 is a key cell cycle regulator that is downregulated in several kinds of human cancers, including gastric cancer. The IVS1 -27G/A polymorphism of KLF6 has been investigated, which can influence susceptibility to gastric cancer and disease outcome. In order to investigate whether the IVS1 -27G/A polymorphism of KLF6 is associated with individual susceptibility to gastric cancer in Korea, the frequency of the polymorphism was examined in 264 gastric cancer patients and 299 healthy controls. Single nucleotide polymorphism (SNP) analysis was performed by amplifying intron 1 of KLF6 and sequencing the products. The frequencies of genotypes: G/G, G/A and A/A were 91.7% (242/264), 5.7% (15/264) and 2.6%, respectively, in gastric cancer cases and 91.9%, 7.0% and 1.1%, respectively, in healthy controls. Genotype frequencies in Korean population were very similar to those of Caucasian population. Interestingly, the male gastric cancer patients showed a significantly higher proportion of the G allele (Chi-Square test, P=0.005) compared to female gastric cancer patients. However, the polymorphism was statistically not associated with increased risk of gastric cancer in Korea. When stratified by histological subtype of gastric cancer, the risk was also not statistically significant. Thus, our results suggested that the IVS1 -27G/A polymorphism of KLF6 is not associated with an increased risk for gastric cancer in Korean population.

No association of MDM2 T309G polymorphism with susceptibility to Korean gastric cancer patients.

Mouse double minute 2 (Mdm2) acts as a negative regulator of p53 by binding to the amino-terminus of p53. The common T309G polymorphism of Mdm2 has been the most frequently investigated, which can influence in cancer susceptibility and disease outcome. The specific aim of this study is to investigate whether the T309G polymorphism of Mdm2 was associated with individual susceptibility to gastric cancer in Korea. The frequency of the polymorphism was examined in 239 gastric cancer patients and 299 healthy controls. Polymorphism analysis was performed by amplifying the first intron of the Mdm2 and digesting with restriction enzyme and sequencing the products. The frequencies of genotypes: T/T, T/G and G/G were 26.8% (64/239), 46.0% (110/239) and 27.2% (65/239), respectively, in gastric cancer cases and 20.4% (61/299), 50.8% (152/ 299) and 28.8% (86/299), respectively, in healthy controls. Statistically, there was no significant difference in the frequency of genotype and allele between healthy control and gastric cancer patients. Finally, the polymorphism was not associated with increased risk of gastric cancer in this population. When stratified by histological subtype of gastric cancer, the risk was also not statistically significant. Our findings suggested that the T309G polymorphism of Mdm2 was not associated with an increased risk for gastric cancer in Korean population.

Chk1 frameshift mutation in sporadic and hereditary non-polyposis colorectal cancers with microsatellite instability.

AIM: Protein kinase Chk1 (hChk1) is essential in human cells for cell cycle arrest in response to DNA damage, and has been shown to play an important role in the G2/M checkpoint. The BRAF mutations have been suggested to be linked with defective mismatch repair in colorectal cancers. The aim of this study was to investigate whether a frameshift mutation within the Chk1 gene contribute to the development or progression of eastern sporadic and hereditary non-polyposis colorectal cancer (HNPCC) with microsatellite instability (MSI). METHODS: We analyzed MSI using the 6 microsatellite markers and a frameshift mutation in the BRAF gene and in poly(A)9 within the Chk1 gene in 51 sporadic colorectal cancer and 14 HNPCC specimens. RESULTS: Eleven of the 51 sporadic colorectal cancers and all of the 14 HNPCCs were MSI-positive. Chk1 frameshift mutations were observed in 2 and 3 sporadic colon cancers and HNPCC, respectively, whereas no BRAF mutations were detected in these samples. Interestingly, all cases with the Chk1 frameshift mutation had high-frequency MSI. CONCLUSION: These results suggest that the Chk1 gene is a target of genomic instability in MSI-positive colorectal cancers and that the Chk1 framshift mutations might be involved in colorectal tumourigenesis through a defect in response to DNA damage in a subset of sporadic colorectal cancers and HNPCCs.

Association of cyclin D1 G870A polymorphism with susceptibility to gastric cancers in Korean male patients.

Cyclin D1 is a key cell cycle regulator that is upregulated in gastric cancer. The common G870A polymorphism of cyclin D1 which can influence cancer susceptibility and disease outcome has been the most frequently investigated. The specific aim of this study is to investigate whether the G870A polymorphism of cyclin D1 was associated with individual susceptibility to gastric cancer in Korea. The frequency of the polymorphism was examined in 253 gastric cancer patients and 442 healthy controls. Polymorphism analysis was performed by amplifying exon 4 of cyclin D1 and sequencing the products. The frequencies of genotypes: G/G, G/A and A/A were 28.1% (71/253), 49.4% (125/253) and 22.5% (57/253), respectively, in gastric cancer cases, and 23.1%, 51.1% and 25.8%, respectively, in healthy controls. Statistically, the polymorphism was not associated with increased risk of gastric cancer. When stratified by histological subtype of gastric cancer, the risk was also not statistically significant. However, the male gastric cancer patients showed a significantly higher proportion of the homozygous G/G genotype and the G allele (Chi-Square test, P = 0.0242 & P = 0.0307) compared to males in the control group. Thus, our findings suggested that the G870A polymorphism of cyclin D1 was not associated with an increased risk for gastric cancer in this population, however, it may contribute to susceptibility to gastric cancer in men.

Saturated molecular map of the rice genome based on an interspecific backcross population.

A molecular map has been constructed for the rice genome comprised of 726 markers (mainly restriction fragment length polymorphisms; RFLPs). The mapping population was derived from a backcross between cultivated rice, Oryza sativa, and its wild African relative, Oryza longistaminata. The very high level of polymorphism between these species, combined with the use of polymerase chain reaction-amplified cDNA libraries, contributed to mapping efficiency. A subset of the probes used in this study was previously used to construct an RFLP map derived from an inter subspecific cross, providing a basis for comparison of the two maps and of the relative mapping efficiencies in the two crosses. In addition to the previously described PstI genomic rice library, three cDNA libraries from rice (Oryza), oat (Avena) and barley (Hordeum) were used in this mapping project. Levels of polymorphism detected by each and the frequency of identifying heterologous sequences for use in rice mapping are discussed. Though strong reproductive barriers isolate O. sativa from O. longistaminata, the percentage of markers showing distorted segregation in this backcross population was not significantly different than that observed in an intraspecific F2 population previously used for mapping. The map contains 1491 cM with an average interval size of 4.0 cM on the framework map, and 2.0 cM overall. A total of 238 markers from the previously described PstI genomic rice library, 250 markers from a cDNA library of rice (Oryza), 112 cDNA markers from oat (Avena), and 20 cDNA markers from a barley (Hordeum) library, two genomic clones from maize (Zea), 11 microsatellite markers, three telomere markers, eleven isozymes, 26 cloned genes, six RAPD, and 47 mutant phenotypes were used in this mapping project. Applications of a molecular map for plant improvement are discussed.

Genetic analysis of the LKB1/STK11 gene in hepatocellular carcinomas.

The tumour suppressor gene, LKB1/STK11, has been mapped to chromosome 19p13, a region showing frequent allelic loss in various human cancers, including hepatocellular carcinoma (HCC). Additionally, LKB1 physically associates with p53 and regulates p53-dependent apoptotic pathways. To investigate whether genetic alterations of LKB1 could be involved in the tumorigenesis of HCC, we analysed the genetic alterations of the LKB1 and p53 genes in seven dysplastic nodules and 80 HCCs. We found one LKB1 missense mutation, CCG-->CTG (Pro-->Leu) at codon 281 within the kinase domain. We also found allelic loss in six of 27 (22%) informative HCC cases and all of them were HBV-positive cases. In addition, we detected seven missense, one nonsense and one silent mutations (nine of 80, 11%) of p53 in HCCs only. These results suggest that genetic alterations of the LKB1 or p53 genes may play an important role in tumour development or progression of a sub-set of HCCs, and may also provide alternative mechanisms to protect the HCC cell from p53-dependent apoptosis.

A neuron-specific gene transfer by a recombinant defective Sindbis virus.

We examined the possibility that Sindbis virus, an alpha virus with a single-stranded RNA genome, would be applied for neuronal gene transfer. The recombinant defective Sindbis viruses were constructed by replacing the structural genes of Sindbis virus with genes encoding beta-galactosidase (rdSind-lacZ) or enhanced green fluorescent protein (rdSind-EGFP). In neuron-glia cocultures prepared from the neocortex, hippocampus, and striatum, EGFP or beta-galactosidase was expressed selectively in neurons 24 h after infection with rdSind-EGFP or rdSind-lacZ. Most cortical neurons were infected with rdSind-lacZ at a multiplicity of infection (M.O.I.) of 5 while glial cells were little infected. In addition, transient neuron-specific expression of beta-galactosidase was observed near injection sites over the next 3 d following administration of rdSind-lacZ in adult rat. In the cortical neurons infected with rdSind-EGFP, treatment with NMDA induced neuritic blebs and cell body swelling in a Na+-dependent manner. Therefore, recombinant defective Sindbis viruses can be used as an efficient and selective vector for gene transfer into neurons and applied to investigate biological role of target genes delivered into neurons in vitro and in vivo.

A case of 49,XXXXX in which the extra X chromosomes were maternal in origin.

This report describes an 11 month old female baby with features of pentasomy X. A molecular and cytogenetic evaluation revealed that her karyotype was 49,XXXXX and her extra X chromosomes were of maternal origin. She has muscular hypotonia, mental retardation, a cleft palate, mild hydrocephalus as a result of dilatation of both lateral ventricles, hyperextensible elbow joints, proximal radioulnar synostosis, clinodactyly of the fifth finger, valgus of the feet, and small hands and feet. In addition, she has a persistent pupillary membrane and congenital chorioretinal atrophy. The pathogenesis of pentasomy X is not clear at present, but it is thought to be caused by successive maternal non-dysjunctions.

Characterization of two rice MADS box genes that control flowering time.

Plants contain a variety of the MADS box genes that encode regulatory proteins and play important roles in both the formation of flower meristem and the determination of floral organ identity. We have characterized two flower-specific cDNAs from rice, designated OsMADS7 and OsMADS8. The cDNAs displayed the structure of a typical plant MADS box gene, which consists of the MADS domain, I region, K domain, and C-terminal region. These genes were classified as members of the AGL2 gene family based on sequence homology. The OsMADS7 and 8 proteins were most homologous to OM1 and FBP2, respectively. The OsMADS7 and 8 transcripts were detectable primarily in carpels and also weakly in anthers. During flower development, the OsMADS genes started to express at the young flower stage and the expression continued to the late stage of flower development. The OsMADS7 and 8 genes were mapped on the long arms of the chromosome 8 and 9, respectively. To study the functions of the genes, the cDNA clones were expressed ectopically using the CaMV 35S promoter in a heterologous tobacco plant system. Transgenic plants expressing the OsMADS genes exhibited the phenotype of early flowering and dwarfism. The strength of the phenotypes was proportional to the levels of transgene expression and the phenotypes were co-inherited with the kanamycin resistant gene to the next generation. These results indicate that OsMADS7 and 8 are structurally related to the AGL2 family and are involved in controlling flowering time.

Sequencing and chromosomal localization of the RA138 gene encoding a rice allergenic protein.

A cDNA clone (RA138) encoding a rice allergenic (RA) protein has been isolated during a large-scale random sequencing of a cDNA library prepared from developing seeds. The nucleotide sequence of the RA138 gene contained an open reading frame (ORF, 477 bp) encoding a 17 kDa protein. The amino acid sequence deduced from the ORF was composed of 159 amino acid residues and was highly homologous to those from RA genes previously isolated, such as RA5 (92% identity), RA14 (73%), and RA17 (68%). The protein contained 10 cysteine residues that were conserved in the alpha-amylase/trypsin inhibitor family including RA proteins. Excluding a putative signal peptide consisting of 26 amino acid residues, the mature protein would be 14.4 kDa in size and have a pI of 7.0. DNA gel blot analysis under high stringency conditions indicated that multiple copies of the RA138 gene were present in the rice genome. The chromosomal location of the RA138 gene has been identified on chromosome 7 in a segregation analysis using a population of 164 recombinant inbred lines derived from a cross between Milyang 23 and Gihobyeo. The locus that may contain multiple copies of the RA138 was located between RFLP markers RG477A and C492 with genetic distances of 10.7 cM and 6.7 cM, respectively.

Construction of hepatitis C-SIN virus recombinants with replicative dependency on hepatitis C virus serine protease activity.

An in vivo assay system was developed for the serine protease of hepatitis C virus (HCV) using the sindbis (SIN) viral replication system in which HCV serine protease activity is essential for the replication of the HCV-SIN chimeric virus. Two chimeric viral cDNA clones were constructed by inserting the NS3/4A region and NS3/4A region with the putative helicase deleted, into the N-terminal region of SIN core protein. The constructs were named Tpro CT and Tpro T, respectively. BHK-21 cells transfected with the in vitro transcribed RNAs from Tpro CT and Tpro T showed specific cytopathic morphology and produced chimeric viruses, Vpro CT and Vpro T. In contrast, in vitro transcribed RNAs from Tpro CTI and Tpro TI, in which serine of catalytic triad of HCV protease was changed to alanine, were not infectious. When the chimeric viruses were passaged in BHK-21 cells at about 0.1 multiplicity of infection (MOI), Vpro T, but not Vpro CT, stably expressed HCV protease for up to five passages. Surprisingly, the cell culture media of BHK-21 cells infected with Vpro T, compared to wild-type sindbis virus, showed rapid pH changes by more than 0.8 pH degree at 72 h post-infection. HCV-SIN hybrid viruses could be used in screening the HCV protease-inhibitor in cell culture systems.

In vivo assay for hepatitis C viral serine protease activity using a secreted protein.

Hepatitis C virus (HCV) is a major pathogen of community-acquired and post-transfusional non-A, non-B hepatitis. Since an in vitro replication system is not available, it is crucial to develop an efficient and sensitive assay system for screening inhibitors of HCV. The fact that the activity of HCV NS3 protease is responsible for the maturation of the nonstructural proteins and viral replication, suggests that NS3 protease is a suitable target for anti-HCV drug development. To devise an assay system in cell culture, we constructed NS3/4A-SEAP (secreted alkaline phosphatase) chimeric gene, in which the SEAP gene was fused in-frame to downstream of NS4A/4B cleavage site. In this system, the SEAP would be secreted into the extracellular media depending on the cleavage activity of the NS3 protease. Our results demonstrate that the NS3/4A-SEAP expression vector encoding wild type NS3 protease, but not mutant NS3 protease, could produce high SEAP activity in the media of both transfected cells and stable expression cell lines. Since the activity of SEAP in the culture media can be monitored quantitatively and continuously by the chemiluminescent method, this assay system will be useful for screening potential inhibitors of HCV protease.

Treatment of metal-contaminated water and vertical distribution of metal precipitates in an upflow anaerobic bioreactor.

A lab-scale upflow anaerobic bioreactor filled with granular sludge and cow manure was operated for 140 days to determine the mechanism of metal removal and the vertical distribution of metal precipitates. Heavy metal ions were removed in the order of Cu2+, Cd2+, Zn2+, Fe2+ and Mn2+ with respect to the height in the reactor. The solid phase analysis showed that the heavy metals were mostly precipitated in the form of metal sulfides by sulfate reduction The contents of metal precipitates in the reactor were as follows: (i) Cd and Zn were highest in the bottom, (ii) Fe was highest at the low-middle layer, and (iii) Mn was increased with the height in the reactor. The vertical distribution of metal sulfides in the reactor was directly related to the solubility product (Ksp). Results obtained in this study suggest a feasibility of the application to separate precipitation metal-containing wastewater.

Reference intervals for whole blood viscosity using the analytical performance-evaluated scanning capillary tube viscometer.

OBJECTIVES: This study was performed to establish the reference intervals for whole blood viscosity (WBV) using the analytical performance-evaluated scanning capillary tube viscometer (SCTV). DESIGN AND METHODS: The analytical performance of the SCTV was evaluated using three different levels of QC materials and sixty human EDTA-blood samples. To establish the reference intervals for WBV, 297 healthy individuals (123 men and 174 women) were selected from 1083 subjects. RESULTS: Within-day precisions with QC materials and human whole blood and between-day precisions with QC materials were below 5.0%, 6.6% and 8.0% in CVs at all shear rates, respectively. Comparison tests between the SCTV and the Brookfield viscometer showed a significant correlation (R(2)=0.972, p<0.001). The reference intervals for WBV in healthy men were 3.66-5.41cP at 300s(-1) and 23.15-36.45cP at 1s(-1) while those in women were 3.27-4.32cP at 300s(-1) and 18.20-27.36cP at 1s(-1), respectively. CONCLUSIONS: Using the analytical performance-evaluated SCTV, the reference intervals for WBV were established in healthy adults, which could be beneficial to the clinical utility of WBV in the aspect of appropriate modalities for the improvement of blood viscosity.

The relationship between body fat mass and erectile dysfunction in Korean men: Hallym Aging Study.

The aim of this study was to assess the relationship between body fat mass (BFM) and erectile dysfunction (ED) in Korean men. This study was a cross-sectional study using data on 208 men (the mean age=67.4+/-8.2). ED was diagnosed by the International Index of Erectile Function (IIEF)-5 and body fat percentage (BF%) was quantified with bioelectrical impedance. BF% was divided into quintiles (quintile 1: < or =20.5%, quintile 2: 20.6-23.2%, quintile 3: 23.3-25.8%, quintile 4: 25.9-28.8%, quintile 5: > or =28.9%). Using subjects with quintile 3 of BF% as reference, the adjusted odds ratios of subjects with the lowest quintile of BF% and with the highest quintile were 9.29 (95% CI: 2.29-37.72) and 4.99 (95% CI: 1.37-18.09), respectively. This study showed that BFM and ED had a U-shaped relationship in Korean men. These findings suggest that not only obesity but also a low BFM may be a risk factor of ED in Asians.

Influence of phenol on biodegradation of p-nitrophenol by freely suspended and immobilized Nocardioides sp. NSP41.

The effect of the presence of an alternate toxic compound (phenol) on the p-nitrophenol (PNP)-degrading activity of freely suspended and calcium alginate immobilized Nocardioides sp. NSP41 was investigated. In the single substrate experiments, when the concentration of phenol and PNP was increased to 1400 mg l(-1) and 400 mg l(-1), respectively, the initial cell concentrations in the freely suspended cell culture should be higher than 1.5 g dry cell weight l(-1) for complete degradation. In the simultaneous degradation experiment, when the initial concentration of phenol was increased from 100 to 400 mg l(-1), the specific PNP degradation rate at the concentration of 200 mg l(-1) was decreased from 0.028 to 0.021 h(-1). A freely suspended cell culture with a high initial cell concentration resulted in a high volumetric degradation rate, suggesting the potential use of immobilized cells for simultaneous degradation. In the immobilized cell cultures, although simultaneous degradation of PNP and phenol was maintained, the specific PNP and phenol degradation rate decreased. However, a high volumetric PNP and phenol degradation rate could be achieved by immobilization because of the high cell concentration. Furthermore, when the immobilized cells were reused in the simultaneous degradation of PNP and phenol, they did not lose their PNP- and phenol-degrading activity for 12 times in semi-continuous cultures. Taken together, the use of immobilized Nocardioides sp. NSP41 for the simultaneous degradation of PNP and phenol at high concentrations is quite feasible because of the high volumetric PNP and phenol degradation rate and the reusability of immobilized cells.