Investigating genome wide DNA methylation in bronchial and lung fibroblasts from healthy individuals and individuals with COPD

Rationale: Lung fibroblasts are implicated in respiratory disease pathology including chronic obstructive pulmonary disease (COPD). Phenotypic differences between fibroblasts isolated from the bronchi versus the lung parenchyma have been described but no studies have compared the cell types on a genome wide scale. DNA methylation is a reversible modification of the DNA structure with the ability to affect cell function via the alteration of gene expression. Here we compared genome wide DNA methylation profiles from bronchial and lung fibroblasts and assessed modification to these profiles in cells isolated from individuals with COPD. Methods: DNA was isolated from lung (LgF) and bronchial fibroblasts (BrF) at passage 4 and bisulphite treated. Site specific, quantitative genome wide methylation was determined using the Illumina 450K Infinium Methylation BeadChip array. Linear modelling and DMRcate functions identified differentially methylated sites and regions respectively between BrF and LgF and from cells isolated from healthy individuals versus those with COPD. Results: 3980 CpG (methylation) sites significantly differed, following Bonferroni correction, between BrF and LgF isolated from healthy individuals. These sites had a broad distribution of effect size, with 240 CpG sites displaying a difference in methylation of >50%. 78 of these sites were validated in a second cohort of matched BrF and LgF isolated from the same individuals. There was genomic proximity to these sites and DMRcate was used to refine the individual CpG sites to 5 regions of interest associated with 5 genes; HLX, TWIST1, CREB5, SKAP2 and PRDM16. Differences in methylation were less pronounced when comparing cells isolated from healthy individuals to those with COPD. In BrF 47 DMRcate regions were identified with a maximum difference in methylation of at least 20%. In LgF 3 DMRcate regions were identified with a maximum difference in methylation of at least 20%. Conclusions: DNA methylation profiles are significantly different between BrF and LgF but only small modifications are associated with COPD. Future work will focus on validating a methylation based marker of lung versus bronchial fibroblasts to differentiate cell types by validating our differential DNA methylation observations with gene/protein expression

Characterization of fracture reservoirs using static and dynamic data: From sonic and 3D seismic to permeability distribution

To characterize the Buena Vista Hills field, the authors have implemented methods of modeling, processing and interpretation. The modeling methods are based on deterministic and stochastic solutions. Deterministic solutions were developed in Phase 1 and applied in Phase 2 to simulate acoustic responses of laminated reservoirs. Specifically, the simulations were aimed at implementing processing techniques to correct P-wave and S-wave velocity logs for scattering effects caused by thin layering. The authors are also including a summary of the theory and the processing steps of this new method for predicting intrinsic dispersion and attenuation in Section 2. Since the objective for correcting velocity scattering effects is to predict intrinsic dispersion from velocity data, they are presenting an application to illustrate how to relate permeability anisotropy with intrinsic dispersion. Also, the theoretical solution for calculating full waveform dipole sonic that was developed in Phase 1 was applied to simulate dipole responses at different azimuthal source orientations. The results will be used to interpret the effects of anisotropy associated with the presence of vertical fractures at Buena Vista Hills. The results of the integration of core, well logs, and geology of Buena Vista Hills is also given in Section 2. The results of this integration will be considered as the input model for the inversion technique for processing production data. Section 3 summarizes accomplishments. In Section 4 the authors present a summary of the technology transfer and promotion efforts associated with this project. In the last section, they address the work to be done in the next six months and future work by applying the processing, modeling and inversion techniques developed in Phases 1 and 2 of this project

Teratology studies of lewisite and sulfur mustard agents: Effects of lewisite in rats and rabbits: Final report: Part 2, Appendices

Lewisite was administered to rats and rabbits by intragastric intubation. Maternal animals were weighed periodically, and, at necropsy (20 dg (days of gestation) in rats and 30 dg in rabbits), were examined for gross lesions of major organs and reproductive performances; live fetuses were weighed and examined for external, internal and skeletal defects. In rats, a dose level of 1.5 mg/kg did not induce toxic or teratogenic responses in maternal animals or their fetuses. At 2.0 mg/kg, 10% maternal mortality, trends in decreased maternal and fetal body weights and a significant reduction in the number of viable fetuses were evident. In rabbit studies, maternal mortality occured in all but one of the lewisite treatment groups and range from 13% to 100% at dose levels of 0.07 and 1.5 mg/kg, respectively. This mortality rate limited the sample size and impaired the detection of statistical significance among treatments. However, at the lowest dose level of the teratology study (0.07 mg/kg), maternal mortality was the only indicator of lewisite toxicity; at the highest dose (0.6 mg/kg), significant findings included 86% maternal mortality, a decrease in maternal body weight gains and an increase in the incidence of fetal stunting, although only a tendency in decreased fetal body weights was observed. These results suggest that maternal mortality was the most important factor in predicting the induction of maternal and fetal effects and, therefore, a ''no observable effect level'' in maternal animals and their fetuses would be between 1.5 and 2.0 mg/kg in rats and less than 0.07 mg/kg in rabbits. Part 2 contains 6 appendices

Dominant lethal study in CD-1 mice following inhalation exposure to 1,3-butadiene: Final technical report

The effects of whole-body inhalation exposures to 1,3-butadiene on the reproductive system was evaluated. The results of dominant lethality in CD-1 male mice that were exposed to 1,3-butadiene are described. Subsequent to exposure, males were mated with two unexposed females. Mating was continued for 8 weeks with replacement of two females each week. Gravid uteri were removed, and the total number, position and status of implantations were determined. The mice were weighed prior to exposure and at 0, 1, 2, 3, 4, 5, 6, 7, and 8 weeks after exposure and at sacrifice. The animals were observed for mortality, morbidity and signs of toxicity throughout the study. 19 refs., 5 figs., 9 tabs

Inhalation developmental toxicology studies: Teratology study of 1,3-butadiene in mice: Final report

Maternal toxicity, reproductive performance and developmental toxicology were evaluated in CD-1 mice following whole-body, inhalation exposures to 0, 40, 200 and 1000 ppM of 1,3-butadiene. The female mice, which had mated with unexposed males were exposed to the chemical for 6 hours/day on 6 through 15 dg and sacrificed on 18 dg. Maternal animals were weighed prior to mating and on 0, 6, 11 and 18 dg; the mice were observed for mortality, morbidity and signs of toxicity during exposure and examined for gross tissue abnormalities at necropsy. Live fetuses were weighed and subjected to external, visceral and skeletal examinations to detect growth retardation and morphologic anomalies. Significant concentration-related decreases were detected in a number of maternal body weight measures. There was a significant concentration-related depression of fetal body weights and placental weights. Body weights of male fetuses of all exposed groups were significantly lower than values for control fetuses; weights of female fetuses were significantly depressed in the mice exposed to 200 and 1000 ppM. In the 200- and 1000-ppM exposure groups, weights of placentas of male fetuses were significantly decreased, but placental weights of female fetuses were significantly affected only in litters exposed to the highest 1,3-butadiene concentration. This exposure regimen produced significant signs of maternal toxicity at concentrations of 200 and 1000 ppM 1,3-butadiene