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Title: The Impact of Roasting Level on the Anticancer Activity of Coffee
Coffee contains high levels of phenolic compounds, including chlorogenic acid and caffeic acid, that have been found to inhibit the growth of cancer cells. Coffee is traditionally prepared by roasting coffee beans which are then ground and brewed in hot water. The degree of roasting could affect the levels of phenolic compounds within the beans and therefore affect their anticancer activity. Additionally, this effect is expected to be the most pronounced in the oral cavity where phenolic compounds are the least degraded. We propose to test the hypothesis that coffee prepared from less roasted beans has greater anticancer effects in oral cancer cells compared with higher roasted coffee. In this project, we will study the effects of roasting level on the anticancer activity of coffee by measuring the growth of human oral cancer cells, and by quantifying specific cellular proteins that are involved in human cancer cell death and survival. The methods used in this project will be mammalian cell culture, biochemical assays, and western blotting. This research may lead to new ways to treat and prevent oral cancer.
Title: Generation and Characterization of Random Mutations of the Active – Site of Lysyl Oxidase
The enzyme lysyl oxidase is a copper – dependent amine oxidase that has been found to play a critical role in many biological systems. The enzyme’s primary role is the cross – linking of collagen and elastin, two proteins that are involved in providing stability and elasticity to connective tissues. More recently, however, lysyl oxidase has been shown to play a paradoxical role in cancer. In the early stages of cancer, a portion of the enzyme, known as the pro – peptide, has been shown to inhibit one of the major pathways leading to cancer. In late stages if cancer, the portion known as the mature enzyme, has been shown to promote metastasis. Efforts in understanding this enzyme have been hampered because there is currently no crystal structure to allow us to understand how the enzyme looks and how it functions.
This project will focus on two aspects of the overall research being carried out in my laboratory.
Goal 1: Generate multiple random mutations in the DNA that makes up the lysyl oxidase gene focusing primarily on the DNA sequence around the active – site of lysyl oxidase and test the effect of these mutations on the enzyme’s ability to carry out its function.
Goal 2: Develop a method by which enzymatic activity tests that are currently being done on a 3.0 mL scale and using fluorescence can be conducted in a microplate format using no more than 0.5 mL of total volume and using absorbance.