A cross-sectional study on the association between vitamin D levels and caries in the permanent dentition of Korean children.

BACKGROUND: A recent study in Canada reported that vitamin D deficiency is associated with dental caries. Because Koreans have been reported to be deficient in vitamin D, we investigated the relationship between dental caries and reduced serum vitamin D levels in Korean children. The purpose of this study was to analyze the relationships between blood vitamin D [25(OH)D] concentrations and dental caries in the permanent dentition of Korean children. METHODS: Data were collected from the Korea National Health and Nutrition Examination Survey performed in 2008-2013. A total of 1688 children (10-12 years of age) were enrolled. Vitamin D intake was measured through analysis of 25-hydroxy vitamin D [25(OH)D] levels. Caries experience in permanent dentition was assessed using the decay-missing-filled teeth (DMFT) index and decayed-missing-filled (DMF) rate. Statistical analyses included complex samples Chi-square tests, complex samples logistic regression analyses, and Pearson's correlations. RESULTS: The group with 25(OH) D levels lower than 50 nmol/L had a higher proportion of children with caries in the permanent dentition and permanent first molar than the group with 25(OH)D levels of 50 nmol/L or more. When external factors, such as sex, were controlled, 25(OH)D levels were not significantly correlated with caries, but were significantly correlated with first molar caries. Children with 25(OH)D levels lower than 50 nmol/L were 1.295 times more likely to have first molar caries than those with 25(OH)D levels of 50 nmol/L or more. Additionally, 25(OH)D levels and DMFT were negatively correlated; however, the degree of correlation was not strong. CONCLUSIONS: The association between 25(OH)D and dental caries is still not clear. However, our findings suggested that vitamin D insufficiency may be a risk factor for dental caries.

Spatiotemporal Evaluation of Water Quality and Hazardous Substances in Small Coastal Streams According to Watershed Characteristics.

In this study, spatial and temporal changes of eight water quality indicators and 30 types of hazardous substances including volatile organic compounds (VOCs), semi-volatile organic compounds (SVOCs), pesticides, and inorganic matters for the small coastal streams along the West Coast of South Korea were investigated. In coastal streams with clear seasonal changes in water quality, larger watershed areas led to greater contamination by particulate matter (i.e., suspended solids, r = 0.89), and smaller watershed areas led to greater contamination by organic matter (i.e., BOD, r = -0.78). The concentration of VOCs and pesticides was higher in agricultural areas, and those of SVOCs and metals were often higher in urban areas. According to the principal component analysis (PCA), during the wet season, the fluctuation in the water quality of coastal streams was higher in urban areas than in agricultural areas. Furthermore, coastal streams in residential areas exhibited higher levels of SVOCs, and those in industrial areas exhibited higher levels of metallic substances. Based on these results, the spatial and temporal trends of water quality and hazardous substances were obtained according to watershed characteristics, thereby clarifying the pollution characteristics of small-scale coastal streams and the major influencing factors.

Crystal structure of bet3 reveals a novel mechanism for Golgi localization of tethering factor TRAPP.

Transport protein particle (TRAPP) is a large multiprotein complex involved in endoplasmic reticulum-to-Golgi and intra-Golgi traffic. TRAPP specifically and persistently resides on Golgi membranes. Neither the mechanism of the subcellular localization nor the function of any of the individual TRAPP components is known. Here, the crystal structure of mouse Bet3p (bet3), a conserved TRAPP component, reveals a dimeric structure with hydrophobic channels. The channel entrances are located on a putative membrane-interacting surface that is distinctively flat, wide and decorated with positively charged residues. Charge-inversion mutations on the flat surface of the highly conserved yeast Bet3p led to conditional lethality, incorrect localization and membrane trafficking defects. A channel-blocking mutation led to similar defects. These data delineate a molecular mechanism of Golgi-specific targeting and anchoring of Bet3p involving the charged surface and insertion of a Golgi-specific hydrophobic moiety into the channels. This essential subunit could then direct other TRAPP components to the Golgi.

Time-resolved resonant elastic soft x-ray scattering at Pohang Accelerator Laboratory X-ray Free Electron Laser.

Resonant elastic x-ray scattering has been widely employed for exploring complex electronic ordering phenomena, such as charge, spin, and orbital order, in particular, in strongly correlated electronic systems. In addition, recent developments in pump-probe x-ray scattering allow us to expand the investigation of the temporal dynamics of such orders. Here, we introduce a new time-resolved Resonant Soft X-ray Scattering (tr-RSXS) endstation developed at the Pohang Accelerator Laboratory X-ray Free Electron Laser (PAL-XFEL). This endstation has an optical laser (wavelength of 800 nm plus harmonics) as the pump source. Based on the commissioning results, the tr-RSXS at PAL-XFEL can deliver a soft x-ray probe (400 eV-1300 eV) with a time resolution of ∼100 fs without jitter correction. As an example, the temporal dynamics of a charge density wave on a high-temperature cuprate superconductor is demonstrated.

The effect of hydrodynamic flow regimes on the algal bloom in a monomictic reservoir.

The effectiveness of a proposed curtain weir to be installed in the transitional zone of a eutrophic reservoir located in monsoon areas on the control of algal blooms in the lacustrine zone where drinking water withdrawals occur was assessed with various hydrodynamic flow regimes. A two-dimensional hydrodynamic and eutrophication model that can accommodate vertical displacement of the weir following the water surface changes was developed and validated using field data obtained from two distinctive hydrological years; drought (2001) and wet (2004). The model adequately reproduced the temporal and spatial variations of temperature, nutrients and phytoplankton concentrations in the reservoir. The efficacy of the curtain weir method found to be diverse for different hydrological conditions and dependent on the inflow densimetric Froude number (Fr(i)). Algal blooming was considerably mitigated by curtailing the transport of nutrients and algae from riverine zone to lacustrine epilimnion zone during the drought year as long as Fr(i) < 1.0. However, some flood events with Fr(i) > 1.0 transported nutrients and algae built upstream of the weir into the downstream euphotic zone by strong entrainments in 2004. Numerical experiments revealed that the efficiency of the weir on the control of algal blooming becomes marginal if the Fr(i) > 3.0.

Water-repellent Hybrid Nanowire and Micro-scale Denticle Structures on Flexible Substrates of Effective Air Retention.

The air retention capability of a superhydrophobic surface plays the crucial role of drag reduction in an aqueous environment. Here, fabrication of water-repellent hybrid structural surfaces by synthesizing superhydrophobic nanowires with a high aspect ratio on micro-scale denticle structures to improve their air holding capacity in water is reported. The hybrid structure is realized by carrying out polymer molding of denticle structures on flexible substrates, hydrothermal growth of nanowires, and subsequent ultra-thin film coating. This technique is readily applicable to large areas, and the fabricated substrates are attachable onto curved surfaces. Our engineered, super water-repellent hybrid structures are found to effectively maintain air bubbles on their surfaces in a highly shear flow condition with a wall shear stress of up to 33.4 Pa, due to the combined effects of the micro-scale denticle structure, which reduces flow resistance, and the superhydrophobic, high-aspect-ratio nanowire structure, which enhances the capillary force to maintain the air bubbles. Our results show the importance of developing superhydrophobic structures of improved air retention capability.

PAL-XFEL soft X-ray scientific instruments and X-ray optics: First commissioning results.

We report an overview of soft X-ray scientific instruments and X-ray optics at the free electron laser (FEL) of the Pohang Accelerator Laboratory, with selected first-commissioning results. The FEL exhibited a pulse energy of 200 µJ/pulse, a pulse width of <50 fs full width at half maximum, and an energy bandwidth of 0.44% at a photon energy of 850 eV. Monochromator resolving power of 10 500 was achieved. The estimated total time resolution between optical laser and X-ray pulses was <270 fs. A resonant inelastic X-ray scattering spectrometer was set up; its commissioning results are also reported.

Vapor phase inclusion of ferrocene and its derivative in a microporous metal-organic porous material and its structural characterization by single crystal X-ray diffraction.

The inclusion of ferrocene and its derivative in metal-organic porous material MOF-5 is achieved by vapor diffusion; single-crystal X-ray diffraction studies using synchrotron radiation of ferrocene-loaded MOF-5 reveal well-ordered guest molecules packed into the pores.

Crystallization and initial crystallographic characterization of the Corynebacterium glutamicum nitrilotriacetate monooxygenase component A.

Safety and environmental concerns have recently dictated the proper disposal of nitrilotriacetate (NTA). Biodegradation of NTA is initiated by NTA monooxygenase, which is composed of two proteins: component A and component B. The NTA monooxygenase component A protein from Corynebacterium glutamicum was crystallized using the sitting-drop vapour-diffusion method in the presence of ammonium sulfate as the precipitant. X-ray diffraction data were collected to a maximum resolution of 2.5 A on a synchrotron beamline. The crystal belongs to the monoclinic space group C2, with unit-cell parameters a = 111.04, b = 98.51, c = 171.61 A, beta = 101.94 degrees . The asymmetric unit consists of four molecules, corresponding to a packing density of 2.3 A(3) Da(-1). The structure was solved by molecular replacement. Structure refinement is in progress.

Asarum sieboldii extracts attenuate growth, acid production, adhesion, and water-insoluble glucan synthesis of Streptococcus mutans.

Asarum sieboldii has been used in traditional folk medicine to treat dental caries and periodontal disease. In the present study, we investigated the inhibitory effect of the ethanol and aqueous extracts of A. sieboldii on the growth, acid production, adhesion, and water-insoluble glucan synthesis of Streptococcus mutans. The ethanol and aqueous extracts of A. sieboldii inhibited the growth and acid production of S. mutans. In the bacterial adherence assay, the ethanol and aqueous extracts of A. sieboldii significantly lowered the adherence of S. mutans. We also found that the ethanol and aqueous extracts of A. sieboldii significantly inhibited the synthesis of water-insoluble glucan by crude glucosyltransferase. These results suggest that A. sieboldii extracts may inhibit the caries-inducing properties of S. mutans. Further studies are necessary to clarify the active constituents of A. sieboldii extracts responsible for such biomolecular activities.

The conserved cis-Pro39 residue plays a crucial role in the proper positioning of the catalytic base Asp38 in ketosteroid isomerase from Comamonas testosteroni.

KSI (ketosteroid isomerase) from Comamonas testosteroni is a homodimeric enzyme that catalyses the allylic isomerization of Delta5-3-ketosteroids to their conjugated Delta4-isomers at a reaction rate equivalent to the diffusion-controlled limit. Based on the structural analysis of KSI at a high resolution, the conserved cis-Pro39 residue was proposed to be involved in the proper positioning of Asp38, a critical catalytic residue, since the residue was found not only to be structurally associated with Asp38, but also to confer a structural rigidity on the local active-site geometry consisting of Asp38, Pro39, Val40, Gly41 and Ser42 at the flexible loop between b-strands B1 and B2. In order to investigate the structural role of the conserved cis-Pro39 residue near the active site of KSI, Pro39 was replaced with alanine or glycine. The free energy of activation for the P39A and P39G mutants increased by 10.5 and 16.7 kJ/mol (2.5 and 4.0 kcal/mol) respectively, while DG(U)H2O (the free-energy change for unfolding in the absence of urea at 25.00+/-0.02 degrees C) decreased by 31.0 and 35.6 kJ/mol (7.4 and 8.5 kcal/mol) respectively, compared with the wild-type enzyme. The crystal structure of the P39A mutant in complex with d-equilenin [d-1,3,5(10),6,8-estrapentaen-3-ol-17-one], a reaction intermediate analogue, determined at 2.3 A (0.23 nm) resolution revealed that the P39A mutation significantly disrupted the proper orientations of both d-equilenin and Asp38, as well as the local active-site geometry near Asp38, which resulted in substantial decreases in the activity and stability of KSI. Upon binding 1-anilinonaphthalene-8-sulphonic acid, the fluorescence intensities of the P39A and P39G mutants were increased drastically, with maximum wavelengths blue-shifted upon binding, indicating that the mutations might alter the hydrophobic active site of KSI. Taken together, our results demonstrate that the conserved cis-Pro39 residue plays a crucial role in the proper positioning of the critical catalytic base Asp38 and in the structural integrity of the active site in KSI.

Structural double-mutant cycle analysis of a hydrogen bond network in ketosteroid isomerase from Pseudomonas putida biotype B.

KSI (ketosteroid isomerase) catalyses an allylic isomerization reaction at a diffusion-controlled rate. A hydrogen bond network, Asp(99).Water(504).Tyr(14).Tyr(55).Tyr(30), connects two critical catalytic residues, Tyr(14) and Asp(99), with Tyr(30), Tyr(55) and a water molecule in the highly apolar active site of the Pseudomonas putida KSI. In order to characterize the interactions among these amino acids in the hydrogen bond network of KSI, double-mutant cycle analysis was performed, and the crystal structure of each mutant protein within the cycle was determined respectively to interpret the coupling energy. The DeltaDeltaG(o) values of Y14F/D99L (Tyr(14)-->Phe/Asp(99)-->Leu) KSI, 25.5 kJ/mol for catalysis and 28.9 kJ/mol for stability, were smaller than the sums (i.e. 29.7 kJ/mol for catalysis and 34.3 kJ/mol for stability) for single mutant KSIs respectively, indicating that the effect of the Y14F/D99L mutation was partially additive for both catalysis and stability. The partially additive effect of the Y14F/D99L mutation suggests that Tyr(14) and Asp(99) should interact positively for the stabilization of the transition state during the catalysis. The crystal structure of Y14F/D99L KSI indicated that the Y14F/D99L mutation increased the hydrophobic interaction while disrupting the hydrogen bond network. The DeltaDeltaG(o) values of both Y30F/D99L and Y55F/D99L KSIs for the catalysis and stability were larger than the sum of single mutants, suggesting that either Tyr(30) and Asp(99) or Tyr(55) and Asp(99) should interact negatively for the catalysis and stability. These synergistic effects of both Y30F/D99L and Y55F/D99L mutations resulted from the disruption of the hydrogen bond network. The synergistic effect of the Y55F/D99L mutation was larger than that of the Y30F/D99L mutation, since the former mutation impaired the proper positioning of a critical catalytic residue, Tyr(14), involved in the catalysis of KSI. The present study can provide insight into interpreting the coupling energy measured by double-mutant cycle analysis based on the crystal structures of the wild-type and mutant proteins.

Association of dental caries with socioeconomic status in relation to different water fluoridation levels.

OBJECTIVES: The aim of this study was to assess the prevalence of dental caries in 11-year-old children, related to water fluoridation and family affluence scale (FAS), as an indicator of socioeconomic status (SES) in Korea. METHODS: A total of eight areas were selected for study: four areas with fluoridated piped water (WF areas) and four areas with nonfluoridated piped water (non-WF areas). Non-WF areas had a similar economic level and population size compared with the WF areas. A total of 1446 elementary school students, 11 years of age, were included. They were examined, and questionnaires completed by their parents were analyzed. In the questionnaire, information about gender, FAS as an indicator of SES, occasions of daily cariogenic snack intake, occasions of daily cariogenic beverage intake, drinking of piped water, cooking with piped water, and usage of oral hygiene supplemental measures were surveyed. The bivariate association between the characteristics of the subjects and the number of decayed, filled, and missing permanent teeth (DMFT score) was analyzed through an independent samples t-test. The difference in the mean DMFT score between different FAS groups was analyzed by DMFT ratio, after adjusting for gender, oral health behaviors, and usage of piped water variables. The DMFT ratio was calculated from a Poisson regression model, because the DMFT score was not normally distributed. RESULTS: There was no significant association between FAS and the mean DMFT score in both areas, by bivariate analysis. After adjusting for each group of confounders, a significant association (95% CI: 1.032-1.513) was found between the FAS and mean DMFT scores in non-WF areas; however, no significant difference was observed in the WF areas (95% CI: 0.766-1.382). CONCLUSIONS: This study supported that water fluoridation could not only lead to a lower prevalence of dental caries, but also help to reduce the effect of SES inequalities on oral health.

Systemic effect of water fluoridation on dental caries prevalence.

OBJECTIVES: The aim of this study was to evaluate the systemic effect of water fluoridation on dental caries prevalence and experience in Cheongju, South Korea, where water fluoridation ceased 7 years previously. METHODS: A cross-sectional survey was employed at two schools where water fluoridation had ceased (WF-ceased area) and at two schools where the water had never been fluoridated (non-WF area). The schools in the non-WF area were of a similar population size to the schools in the WF-ceased area. Children of three age groups were examined in both areas: aged 6 (n = 505), 8 (n = 513), and 11 years (n = 467). The differences in the mean number of decayed or filled primary teeth (dft) and the mean number of decayed, missing, or filled permanent teeth (DMFT) scores between areas after adjusting for oral health behaviors and socio-demographic factors were analyzed by a Poisson regression model. RESULTS: The regression model showed that the DMFT ratio for children aged 11 years in the WF-ceased area was 0.581 (95% CI 0.450-0.751). In contrast, the dft ratio for age 6 in the WF-ceased area was 1.158 (95% CI 1.004-1.335). Only the DMFT ratio for age 8 (0.924, 95% CI 0.625-1.368) was not significant. CONCLUSIONS: While 6-year-old children who had not ingested fluoridated water showed higher dft in the WF-ceased area than in the non-WF area, 11-year-old children in the WF-ceased area who had ingested fluoridated water for approximately 4 years after birth showed significantly lower DMFT than those in the non-WF area. This suggests that the systemic effect of fluoride intake through water fluoridation could be important for the prevention of dental caries.

Methane sorption and structural characterization of the sorption sites in Zn2(bdc)2(dabco) by single crystal X-ray crystallography.

Sorption isotherms of methane in Zn(2)(bdc)(2)(dabco) are measured up to a pressure of 35 bar in the temperature range between 198-296 K. The methane sorption measurements at 296 K showed an uptake of 137 cm(3) cm(-3) at 35 bar. The enthalpy of methane adsorption for Zn(2)(bdc)(2)(dabco) estimated by the virial equation is 13.6 kJ mol(-1) at zero coverage. X-ray structure analysis of methane-adsorbed Zn(2)(bdc)(2)(dabco) by synchrotron radiation at 90 K revealed that methane molecules occupy three independent sorption sites (A, B, and C) with a stoichiometry of Zn(2)(bdc)(2)(dabco) x 6.69 CH(4), which is consistent with the results of the gas sorption measurements at 198 K. In a cavity, eight symmetry-related methane sorption sites A are located near the {Zn(2)(CO(2))(4)} paddle-wheel units, while four symmetry-related methane sorption sites B are near the center of the small windows along the a and b axes. Both A and B sites are half-occupied. Methane molecules occupying sites A are not only in van der Waals contact with the paddle-wheel units, but also interact with the phenyl rings of bdc ligands through partial pi-HC interactions. Methane molecules in B sites interact with the side of the phenyl rings through van der Waals interaction. The site C, located at the center of the cavity, is a secondary sorption site; methane molecules occupying sites C are in van der Waals contact with those in sites A and B.

Microporous magnesium and manganese formates for acetylene storage and separation.

Acetylene sorption of microporous metal formates M(HCOO)2 (M = Mg and Mn) was investigated. Measurements of acetylene sorption at 196, 275, and 298 K showed a Type I isotherm with quick saturation at low pressures, and 50-75 cm3 g(-1) uptake at 1.0 atm. The single-crystal X-ray structure analysis of the acetylene-adsorbed metal formates revealed that acetylene molecules occupy two independent positions in the zigzag channels of the frameworks with a stoichiometry of M(HCOO)2 x 1/3C2H2, which is consistent with the gas sorption experiments. No specific interaction except van der Waals interactions between the adsorbed acetylene molecules and the walls of the frameworks was found. Sorption properties of other gases, including CO2, CH4, N2, O2, and H2, were also investigated. When the temperature was increased to 298 K, the amount of adsorbed acetylene was still above 60 cm3 g(-1) for Mg(HCOO)2 and 50 cm3 g(-1) for Mn(HCOO)2, whereas the uptake of other gases decreased substantially. The microporous metal formates may thus be useful not only for the storage of acetylene but also its separation from other gases at room or slightly higher temperatures.

Structural basis for the extended substrate spectrum of CMY-10, a plasmid-encoded class C beta-lactamase.

The emergence and dissemination of extended-spectrum (ES) beta-lactamases induce therapeutic failure and a lack of eradication of clinical isolates even by third-generation beta-lactam antibiotics like ceftazidime. CMY-10 is a plasmid-encoded class C beta-lactamase with a wide spectrum of substrates. Unlike the well-studied class C ES beta-lactamase from Enterobacter cloacae GC1, the Omega-loop does not affect the active site conformation and the catalytic activity of CMY-10. Instead, a three-amino-acid deletion in the R2-loop appears to be responsible for the ES activity of CMY-10. According to the crystal structure solved at 1.55 A resolution, the deletion significantly widens the R2 active site, which accommodates the R2 side-chains of beta-lactam antibiotics. This observation led us to demonstrate the hydrolysing activity of CMY-10 towards imipenem with a long R2 substituent. The forced mutational analyses of P99 beta-lactamase reveal that the introduction of deletion mutations into the R2-loop is able to extend the substrate spectrum of class C non-ES beta-lactamases, which is compatible with the isolation of natural class C ES enzymes harbouring deletion mutations in the R2-loop. Consequently, the opening of the R2 active site by the deletion of some residues in the R2-loop can be considered as an operative molecular strategy of class C beta-lactamases to extend their substrate spectrum.

Highly efficient protein expression and purification using bacterial hemoglobin fusion vector.

Recently developed bacterial hemoglobin (VHb) fusion expression vector has been widely used for the production of many target proteins due to its distinctive properties of expressing fusion protein with red color which facilitates visualization of the steps in purification, and increasing solubility of the target proteins. However, after intensive use of the vector, several defects have been found. In this report, we present a modified VHb fusion vector (pPosKJ) with higher efficiency, in which most of the defects were eliminated. First, it was found that thrombin protease often digests target protein as well as inserted thrombin cleavage site, so it was replaced by a TEV cleavage site for more specific cleavage of VHb from target protein. Second, a glycine-rich linker sequence was inserted between 6x his-tag and VHb to improve the affinity of 6x his-tag to Ni-NTA resin, resulting in higher purity of eluted fusion protein. Third, EcoRI and XhoI restriction sites located elsewhere in the vector were removed to make these restriction sites available for the cloning of target protein coding genes. A pPosKJ vector was fully examined with an anti-apoptotic BCL-2 family member of Caenorhabditis elegans, CED-9. A C-terminal VHb fusion expression vector (pPosKJC) was also constructed for stable expression of target proteins that may be difficult to express with an N-terminal fusion. Vaccinia-related kinase 1 (VRK1) was also successfully expressed and purified using the vector with high yield. Taken together, we suggest that the VHb fusion vector may be well suited for high-throughput protein expression and purification.

The inverted cucurbit[n]uril family.

We report the isolation, characterization, and recognition behavior of iCB[6] and iCB[7], which are diastereomers of CB[6] and CB[7], respectively, containing a single inverted glycoluril unit. Product resubmission experiments establish that these inverted CB[n] are intermediates in the mechanism of CB[n] formation. As a consequence of the inverted glycoluril ring, these inverted cucurbiturils possess a permanent dipole moment, are slightly smaller than their diastereomers, show distinctive selectivity in their recognition behavior, and report directly on the contents of their hydrophobic cavity.