Quantitative Analysis of Diepoxybutane Damage within the Chicken β-globin Domain

In industrial polymer and synthetic rubber production facilities, workers are exposed to 1,3-butadiene. This compound is converted in vivo to 1,2,3,4-diepoxybutane (DEB) and has been linked to increased incidences of cancer in these individuals. Carcinogenesis has been attributed to formation of DEB induced DNA interstrand cross-links. Previous studies have demonstrated that DEB cross-links deoxyguanosine residues within 5'-GNC sequences in synthetic DNA, in restriction fragments, and in defined sequence nucleosomes. The current study utilized the polymerase chain reaction (PCR) to examine DEB damage frequencies within nuclear genes, found within "open" regions of chromatin, as compared to regions of unexpressed sequence that reside in tightly packed, "closed" chromatin, to more closely model DEB reactivity in vivo. These initial studies have been performed in chicken liver homogenates. Preliminarily, we have found a dose-dependent DEB lesion-forming response within "open" chromatin. DEB appears to have little-to-no effect upon regions of "closed" chromatin.

Mapping of Diepoxybutane Damage in the Cytochrome b Domain of Mitochondrial DNA and the β-globin Domain of Nuclear Chicken DNA Using Quantitative PCR

Diepoxybutane (DEB), a known industrial carcinogen, reacts with DNA primarily at the N7 position of deoxyguanosine residues and creates interstrand cross-links at the sequence 5'-GNC. Since N7-N7 cross-links cause DNA to fragment upon heating, quantative polymerase chain reaction (QPCR) is being used in this experiment to measure the amount of DEB damage (lesion frequency) with three different targets-mitochondrial (unpackaged), open chromatin region, and closed chromatin region. Initial measurements of DEB damage within these three targets were not consistent because the template DNA was not the limiting reagent in the PCR. Follow-up PCR trials using a limiting amount of DNA are still in progress although initial experimentation looks promising. Sequencing of these three targets to confirm the primer targets has only been successfully performed for the closed chromatin target and does not match the sequence from NIH used to design that primer pair. Further sequencing trials need to be conducted on all three targets to assure that a mitochondrial, open chromatin, and closed chromatin region are actually being amplified in this experimental series.

Beyond the Threshold: Life in the New Russia

Amanda Sprang spent nine months, from September of 1995 to May of 1996, studying at Colby College's program in St. Petersburg, Russia. Through contacts made during previous trips to Russia in middle and high school, Amanda was able to quickly rekindle her old friendships and make new ones with many young Russians from different backgrounds. The following work is a collection of twelve essays about life in the New Russia. The essays are framed by a foreword and an epilogue that help place the entire work in a historical context. Although the theme of each essay emerges from a particular incident, within every story Amanda has addressed numerous topics relating to Russian life in today’s changing society. Her first essay, “Art Klinika," takes place in an avant-garde night club in St. Petersburg, and includes a brief yet impressionable, encounter with three young Russian men. “The Birthday Party” recalls a wild evening at the home of her close friend, showing how the Russians greet special occasions. Both the third and fourth essays take place in Moscow, where Amanda returns to visit old friends. These two essays portray the lives of the new economic elite in comparison with the average citizen, as well as show how young Russians face the new challenges that greet them. "Politics Russian Style" recalls a political rally in St. Petersburg, and attempts to shed light on the wacky political world of an infant democracy. Chapters Six through Ten take place away from the western cities of St. Petersburg and Moscow, as Amanda brings us to the cold, mysterious land of Siberia in the dead of winter. She recounts her five day train ride with a retired, high-powered, Communist party official, her experiences in the provincial city of Irkutsk, and a brief trip to a Buddhist monastery and, later, an excursion to Lake Baikal. Back in St. Petersburg, Chapter Eleven gives a humorous account of a ski trip with several Russian friends. Amanda finishes her work with her final chapter, “The Dacha," which describes a weekend spent at a Russian country home with her friend's family.