The early UL31 gene of equine herpesvirus 1 encodes a single-stranded DNA-binding protein that has a nuclear localization signal sequence at the C-terminus.

The amino acid sequence of the UL31 protein (UL31P) of equine herpesvirus 1 (EHV-1) has homology to that of the ICP8 of herpes simplex virus type 1 (HSV-1). Here we show that the UL31 gene is synergistically trans-activated by the IEP and the UL5P (EICP27). Detection of the UL31 RNA transcript and the UL31P in EHV-1-infected cells at 6h post-infection (hpi) as well as metabolic inhibition assays indicated that UL31 is an early gene. The UL31P preferentially bound to single-stranded DNA over double-stranded DNA in gel shift assays. Subcellular localization of the green fluorescent protein (GFP)-UL31 fusion proteins revealed that the C-terminal 32 amino acid residues of the UL31P are responsible for the nuclear localization. These findings may contribute to defining the role of the UL31P single-stranded DNA-binding protein in EHV-1 DNA replication.

Dietary patterns and metabolic syndrome risk factors among adolescents.

PURPOSE: Unbalanced diets and decreased physical activity have contributed to increased prevalence of obesity and metabolic syndrome in adolescents. We have performed a systematic review and data analysis to examine the association between dietary pattern and metabolic syndrome risk factors in adolescents. METHODS: We searched the PubMed and BioMedLib databases for appropriate articles published during the past 10 years and selected 6 articles. The studies reviewed applied factor analysis or cluster analysis to extract dietary patterns. For data analysis, we examined the association between dietary patterns and the prevalence of metabolic syndrome risk factors using data of 3,168 adolescents (13 to 18 years) obtained from 4 consecutive Korean Nutrition Health and Nutrition Examination Surveys (1998, 2001, 2005, and 2007 to 2009). RESULTS: Our systematic review confirmed that western dietary patterns are positively associated with metabolic syndrome risk factors such as obesity and elevated triglycerides, while traditional dietary patterns were negatively associated. Data analysis found that the number of adolescents aged 16 to 18 years who had "Rice & Kimchi" dietary pattern decreased, while the number having western dietary patterns increased during the 1998 to 2009 time frame. There were no changes in the dietary patterns in adolescents aged 13 to 15 years. The risk of elevated serum triglycerides and reduced serum high density lipoprotein cholesterol was high in the "Rice & Kimchi" dietary pattern compared to the other dietary pattern groups. CONCLUSION: Because adolescents' dietary patterns are changing continuously and have long-term effects, further studies on the dietary patterns of adolescents and their health effects into adulthood are necessary.

Income-related health inequalities across regions in Korea.

INTRODUCTION: In addition to economic inequalities, there has been growing concern over socioeconomic inequalities in health across income levels and/or regions. This study measures income-related health inequalities within and between regions and assesses the possibility of convergence of socioeconomic inequalities in health as regional incomes converge. METHODS: We considered a total of 45,233 subjects (≥ 19 years) drawn from the four waves of the Korean National Health and Nutrition Examination Survey (KNHANES). We considered true health as a latent variable following a lognormal distribution. We obtained ill-health scores by matching self-rated health (SRH) to its distribution and used the Gini Coefficient (GC) and an income-related ill-health Concentration Index (CI) to examine inequalities in income and health, respectively. RESULTS: The GC estimates were 0.3763 and 0.0657 for overall and spatial inequalities, respectively. The overall CI was -0.1309, and the spatial CI was -0.0473. The spatial GC and CI estimates were smaller than their counterparts, indicating substantial inequalities in income (from 0.3199 in Daejeon to 0.4233 Chungnam) and income-related health inequalities (from -0.1596 in Jeju and -0.0844 in Ulsan) within regions.The results indicate a positive relationship between the GC and the average ill-health and a negative relationship between the CI and the average ill-health. Those regions with a low level of health tended to show an unequal distribution of income and health. In addition, there was a negative relationship between the GC and the CI, that is, the larger the income inequalities, the larger the health inequalities were. The GC was negatively related to the average regional income, indicating that an increase in a region's average income reduced income inequalities in the region. On the other hand, the CI showed a positive relationship, indicating that an increase in a region's average income reduced health inequalities in the region. CONCLUSION: The results suggest that reducing health inequalities across regions require a more equitable distribution of income and a higher level of average income and that the higher the region's average income, the smaller its health inequalities are.

The early UL3 gene of equine herpesvirus-1 encodes a tegument protein not essential for replication or virulence in the mouse.

The UL3 gene of equine herpesvirus-1 (EHV-1) is retained in the genome of defective interfering particles and encodes a ~33kDa myristylated protein. Further characterization showed that the UL3 gene is trans-activated only by the sole immediate early (IE) protein and encodes an early protein that is dispensable for EHV-1 replication and localizes in the tegument of purified virions. UL3-deleted EHV-1 (vL11ΔUL3) exhibits properties of host cell tropism, plaque size, and growth kinetics similar to those of the parental virus. Expression levels of EHV-1 proteins representative of all three gene classes in vL11ΔUL3-infected cells were identical to those in cells infected with parental virus. Mice intranasally infected with vL11ΔUL3 and parental virus showed no significant difference in mortality or virus lung titers. These findings suggest that the UL3 protein does not play a major role in the biology of EHV-1 in cell culture or virulence in the mouse.

The equine herpesvirus-1 (EHV-1) IR3 transcript downregulates expression of the IE gene and the absence of IR3 gene expression alters EHV-1 biological properties and virulence.

The IR3 transcript of equine herpesvirus-1 (EHV-1) harbors 117 nts antisense to the immediate-early (IE) mRNA, suggesting it plays a regulatory role. Here, we show that the IR3 transcript downregulates IE gene expression and that the absence of IR3 expression altered EHV-1 biological properties and virulence in mice. Reporter assays revealed that the IR3/IE overlapping sequences [IR3(+226/+342)] and an additional IR3(+343/+433) region are necessary for the IR3 RNA to downregulate IE expression. Experiments with the DeltaIR3 EHV-1 showed that the IR3 gene is dispensable for EHV-1 replication. Protein expression of the IE and representative EHV-1 genes was increased in cells infected with DeltaIR3 EHV-1 as compared to that of cells infected with wt EHV-1. The DeltaIR3 EHV-1 exhibited increased virulence in mice as compared to the parent virus. The finding that the IR3 transcript affects IE gene expression extends the role of RNA as a regulatory molecule in alphaherpesvirus infection.

Income-related health inequalities in Korea.

Data from the 2001 Korean National Health and Nutrition Examination Survey and the ill health concentration index (CI) were used to examine income-related health inequalities among Koreans. Participants (>19 years old) were requested to provide information regarding monthly household income, expenditures, subjective living conditions, and health status. Ill health was determined both subjectively through self-rated health (SRH) scores and objectively through the number of diseases (ND). At the individual level, the CIs for SRH and ND were -0.147 and -0.093, respectively; age-gender adjusted CIs were -0.065 and -0.071, respectively. These values remained unchanged when estimating CI for grouped data. These results indicate that ill health was more pronounced among lower income groups in Korea. However, avoidable health inequality in Korea was smaller than in the United Kingdom and the United States, larger than in Sweden, Eastern Germany, Finland, and Western Germany, and roughly equal to the Netherlands, Spain, and Switzerland.

The equine herpesvirus-1 IR3 gene that lies antisense to the sole immediate-early (IE) gene is trans-activated by the IE protein, and is poorly expressed to a protein.

The unique IR3 gene of equine herpesvirus 1 (EHV-1) is expressed as a late 1.0-kb transcript. Previous studies confirmed the IR3 transcription initiation site and tentatively identified other cis-acting elements specific to IR3 such as a TATA box, a 443 base pair 5'untranslated region (UTR), a 285 base pair open reading frame (ORF), and a poly adenylation (A) signal [Holden, V.R., Harty, R.N., Yalamanchili, R.R., O'Callaghan, D.J., 1992. The IR3 gene of equine herpesvirus type 1: a unique gene regulated by sequences within the intron of the immediate-early gene. DNA Seq. 3, 143-152]. Transient transfection assays revealed that the IR3 promoter is strongly trans-activated by the IE protein (IEP) and that coexpression of the IEP with the early EICP0 and IR4 regulatory proteins results in maximal trans-activation of the IR3 promoter. Gel shift assays revealed that the IEP directly binds to the IR3 promoter region. Western blot analysis showed that the IR3 protein produced in E. coli was detected by antibodies to IR3 synthetic peptides; however, the IR3 protein was not detected in EHV-1 infected cell extracts by these same anti-IR3 antibodies, even though the IR3 transcript was detected by northern blot. These findings suggest that the IR3 may not be expressed to a protein. Expression of an IR3/GFP fusion gene was not observed, but expression of a GFP/IR3 fusion gene was detected by fluorescent microscopy. In further attempts to detect the IR3/GFP fusion protein using anti-GFP antibody, western blot analysis showed that the IR3/GFP fusion protein was not detected in vivo. Interestingly, a truncated form of the GFP/IR3 protein was synthesized from the GFP/IR3 fusion gene. However, GFP/IR3 and IR3/GFP fusion proteins of the predicted sizes were synthesized by in vitro coupled transcription and translation of the fusion genes, suggesting poor expression of the IR3 protein in vivo. The possible role of the IR3 transcript in EHV-1 infection is discussed.

Awareness and perceived necessity of dietary guidelines among urban Koreans by gender.

We examined an urban population's awareness of dietary guidelines to determine whether they perceived them to be necessary, and identified the factors affecting this awareness and perceived necessity. Data were collected via physical examinations and face-to-face interviews. Health indicators were collected from health examinations, and information on individual characteristics, including awareness, perceived necessity, and dietary status were obtained from the interviews. The subjects' sociodemographic, health and dietary status were examined as potential factors influencing awareness and perceived necessity. Descriptive statistics, contingency tables, and logistic regression modeling were used in the analysis. Low awareness of the dietary guidelines was displayed by the elderly and poorly educated individuals, and those without a spouse. Most people who were not aware of the dietary guidelines thought the guidelines unnecessary. Males with hypertension and lower hemoglobin levels, and females with less education had low awareness. Elderly females with lower bone density had less perceived necessity. Males and females with poor dietary status reported low perceived necessity.

Predictors of interest in nutrition topics and willingness to participate in local nutrition programmes.

OBJECTIVE: To investigate the predictors of interest in nutrition topics and willingness to participate in weight management courses and nutrition-related chronic disease management courses. METHODS: Data from a cross-sectional survey (n = 1007 Koreans aged >or=30) on nutrition-related conditions, nutrition behaviours, interest in nutrition topics and willingness to participate in two types of nutrition education courses, and general characteristics. Binary logistic regression was used to identify predictors of the participants' interest in the topic and willingness to participate. RESULTS: Younger, richer and more-educated participants were significantly more likely to be willing to participate in 'nutrition-related chronic disease management'. Having a higher income and being male increased the odds of being interested in this topic. Younger, more obese, high-income females and those interested to keep their weight and reduce fatty foods were more likely to be willing to participate in 'weight management'. CONCLUSION: There is a need to increase awareness about the link between nutrition and related health problems, especially regarding 'hidden' health problems to tailor the programmes especially for older, low-income and less-educated people and to focus on increasing their interest in nutrition topics.

Robust production of infectious hepatitis C virus (HCV) from stably HCV cDNA-transfected human hepatoma cells.

Hepatitis C virus (HCV) chronically infects approximately 170 million people worldwide, with an increased risk of developing cirrhosis and hepatocellular carcinoma. The study of HCV replication and pathogenesis has been hampered by the lack of an efficient stable cell culture system and small-animal models of HCV infection and propagation. In an effort to develop a robust HCV infection system, we constructed stable human hepatoma cell lines that contain a chromosomally integrated genotype 2a HCV cDNA and constitutively produce infectious virus. Transcriptional expression of the full-length HCV RNA genome is under the control of a cellular Pol II polymerase promoter at the 5' end and a hepatitis delta virus ribozyme at the 3' end. The resulting HCV RNA was expressed and replicated efficiently, as shown by the presence of high levels of HCV proteins as well as both positive- and negative-strand RNAs in the stable Huh7 cell lines. Stable cell lines robustly produce HCV virions with up to 10(8) copies of HCV viral RNA per milliliter (ml) of the culture medium. Subsequent infection of naïve Huh7.5 cells with HCV released from the stable cell lines resulted in high levels of HCV proteins and RNAs. Additionally, HCV infection was inhibited by monoclonal antibodies specific to CD81 and the HCV envelope glycoproteins E1 and E2, and HCV replication was suppressed by alpha interferon. Collectively, these results demonstrate the establishment of a stable HCV culture system that robustly produces infectious virus, which will allow the study of each aspect of the entire HCV life cycle.

High-level scu-PA production by butyrate-treated serum-free culture of recombinant CHO cell line.

The MGpUK-5 cell line, transformed with a single-chain urokinase-type plasminogen activator (scu-PA) minigene, generated mRNA transcripts and scu-PA titers corresponding to 65% or 86% of the amount generated before serum-free adaptation, despite significant loss of scu-PA gene copies during adaptation to serum-free culture. To further augment scu-PA production, a culture strategy employing sodium butyrate was explored. In 60-mL spinner flask cultures, sodium butyrate in the concentration range 1-10 mM allowed scu-PA production 2- to 3-fold higher than that in the negative control culture. Its productivity-enhancing activity was dependent on cell density in a range of 1-5 x 10(6) cells/mL, generating 72,200 +/- 8,100 IU/mL (480 +/- 50 mg/L) in 60-mL spinner flask cultures. To confirm this result, cells were grown to 4.4 x 10(6) cells/mL and treated with 5 mM sodium butyrate in a 2.5-L perfusion culture. The scu-PA titer increased more than 2-fold, and specific production rate of scu-PA increased 3-fold by this treatment. Overall, this perfusion culture gave rise to 1.7 x 10(8) IU scu-PA (1.1 g), comparable to total scu-PA production in a batch butyrate-treated culture performed at a 25-L bioreactor scale (1.3-3.5 g). Our results suggest that sodium butyrate treatment on high-density culture enables scu-PA production in gram quantities.