Several methods for the in vivo study of angiogenesis are available, and each angiogenic assay presents distinct advantages and disadvantages. In this study, we present a new method for the quantitation of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane (CAM), based on the implantation of gelatin sponges on the top of growing CAM, on day 8 of incubation. After implantation, the sponges were treated with a stimulator (recombinant human basic fibroblast growth factor, FGF2) or an inhibitor (a rabbit polyclonal anti-FGF2 antibody) of blood vessel formation. Blood vessels growing vertically into the sponge and at the boundary between sponge and surrounding CAM mesenchyme were counted by a morphometric method on day 12. In addition, to assess whether the gelatin sponge is an appropriate vehicle to deliver cultured cells and evaluate their angiogenic potential, mouse aortic endothelial cells were cotransfected with human FGF2 and the Escherichia coli β -galactosidase (β -GAL) reporter gene. Stable transfectants were absorbed by the sponge, and evaluation of the angiogenic response was paralleled by β -GAL staining to visualize implanted cells. This technique may facilitate the discovery and development of agonists or antagonists of angiogenesis.
Interindividual variability of antioxidant enzyme activity in healthy young adults was partially explained by significant associations with three known genetic polymorphisms, and was further modified by gender and ethnicity. A substantial component of this variability may be attributable to differences in diet, environmental exposures, and additional genetic factors.
Recent studies have demonstrated widespread pesticide exposures in pregnant
women and in children. Plasma paraoxonase 1 (PON1) plays an important
role in detoxification of various organophosphates. The goals of
this study were to examine in the Center for Health Assessment of Mothers
and Children of Salinas (CHAMACOS) birth cohort of Latina mothers
and their newborns living in the Salinas Valley, California, the frequencies
of five PON1 polymorphisms in the coding region (192QR and 55LM) and the promoter region (−162AG, −909CG, and −108CT) and to determine their associations with PON1 plasma levels [phenylacetate
arylesterase (AREase)] and enzyme activities of
paraoxonase (POase) and chlorpyrifos oxonase (CPOase). Additionally, we
report results of PON1 linkage analysis and estimate the predictive value of haplotypes for PON1 plasma
levels. We found that PON1−909, PON1−108, and PON1192 had an equal frequency (0.5) of both alleles, whereas PON1−162 and PON155 had lower variant allele frequencies (0.2). Nearly complete linkage disequilibrium
was observed among coding and promoter polymorphisms (p < 0.001), except PON1192 and PON1−162 (p > 0.4). Children’s PON1 plasma levels (AREase ranged from 4.3 to 110.7 U/mL) were 4-fold lower than their mothers’ (19.8 to 281.4 U/mL). POase
and CPOase activities were approximately 3-fold
lower in newborns than in mothers. The genetic contribution to PON1 enzyme
variability was higher in newborns (R2 = 25.1% by genotype and 26.3% by haplotype) than
in mothers (R2 = 8.1 and 8.8%, respectively). However, haplotypes and
genotypes were comparable in predicting PON1 plasma levels in mothers
and newborns. Most of the newborn children and some pregnant women in
this Latino cohort may have elevated susceptibility to organophosphate
toxicity because of their PON1192 genotype and low PON1 plasma levels.
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