Special Section on Pediatric Drug Disposition and Pharmacokinetics Role of Chromatin Structural Changes in Regulating Human CYP3A Ontogeny s

ABSTRACT Variability in drug-metabolizing enzyme developmental trajectories contributes to interindividual differences in susceptibility to chemical toxicity and adverse drug reactions, particularly in the first years of life. Factors linked to these interindividual differences are largely unknown, but molecular mechanisms regulating ontogeny are likely involved. To evaluate chromatin structure dynamics as a likely contributing mechanism, age-dependent changes in modified and variant histone occupancy were evaluated within known CYP3A4 and 3A7 regulatory domains. Chromatin immunoprecipitation using fetal or postnatal human hepatocyte chromatin pools followed by quantitative polymerase chain reaction DNA amplification was used to determine relative chromatin occupancy by modified and variant histones. Chromatin structure representing a poised transcriptional state (bivalent chromatin), indicated by the occupancy by modified histones associated with both active and repressed transcription, was observed for CYP3A4 and most 3A7 regulatory regions in both postnatal and fetal livers. However, the CYP3A4 regulatory regions had significantly greater occupancy by modified histones associated with repressed transcription in the fetal liver. Conversely, some modified histones associated with active transcription exhibited greater occupancy in the postnatal liver. CYP3A7 regulatory regions also had significantly greater occupancy by modified histones associated with repressed transcription in the fetus. The observed occupancy by modified histones is consistent with chromatin structural dynamics contributing to CYP3A4 ontogeny, although the data are less conclusive regarding CYP3A7. Interpretation of the latter data may be confounded by celltype heterogeneity in the fetal liver

Residual stresses in weld deposited clad pressure vessels and nozzles

Results of through-thickness residual stress measurements are provided for a variety of samples of weld deposited 308/309L stainless steel and Alloy 600 cladding on low-alloy pressure vessel ferritic steels. Clad thicknesses between 5 and 9mm on samples that vary in thickness from 45 to 200mm were studied. The samples were taken from flat plates, from a spherical head of a pressure vessel, from a ring-segment of a nozzle bore, and from the transition radius between a nozzle and a pressure vessel shell. A layer removal method was used to measure the residual stresses. The effects of uncertainties in elastic constants (Young`s modulus and Poisson`s ratio) as well as experimental error are assessed. All measurements were done at room temperature. The results of this work indicate that curvature plays a significant role in cladding residual stress and that tensile residual stresses as high as the yield stress can be measured in the cladding material. Since the vessel from which the spherical and nozzle corner samples were taken was hydrotested, and the flat plate specimens were taken from specimens used in mechanical fatigue testing, these results suggest that rather high tensile residual stresses can be retained in the cladding material even after some mechanical loading associated with hydrotesting and that higher levels of hydrotest loading would be required to alter the cladding residual stresses

A study of chromosomal aberrations and chromosomal fragility after recombinant growth hormone treatment.

Increased chromosomal rearrangements and chromosomal fragility have been previously observed in lymphocytes of children treated with human GH, implying that treatment could predispose to malignancy. Twenty-four children with classic GH deficiency, neurosecretory GH dysfunction, and Turner syndrome were treated with recombinant human GH (0.3 mg x kg(-1) x wk(-1)). Metaphase cells were assessed for spontaneous chromosomal and chromatid aberrations at baseline and 6 mo into treatment. There were no significant differences in aberrations between baseline and the 6-mo samples. However, the mean frequency of chromatid-type aberrations on a per cell basis was significantly higher than at baseline, 0.0088 versus 0.0064 aberrations per cell (p \u3c 0.024). Two patients contributed inordinately to this increase. A third sample from these two patients was almost identical to their baseline samples. Cells were also irradiated in vitro (3 Gy) to assess chromosomal fragility. After irradiation, no patient showed a significant difference for any aberration type, although there was a significantly lower frequency of ring chromosomes on a per cell basis in the 6-mo samples (p \u3c 0.001). We find no evidence that GH therapy influences spontaneous chromosomal aberrations or chromosomal fragility