Date of Award

2017

Document Type

Open Access Dissertation

Degree Name

Doctor of Philosophy in Biological Sciences (PhD)

Administrative Home Department

Department of Biological Sciences

Advisor 1

Rupali Datta

Advisor 2

Ramakrishna Wusirika

Committee Member 1

Aparna Deshpande

Committee Member 2

Tarun Dam

Abstract

Rice Callus Suspension Culture (RCSC) has been shown to exhibit potent anticancer activity in vitro. In this study we aimed to investigate the anti-inflammatory and immune-modulating properties of RCSC on three inflamed normal colon epithelial cell lines, InEpC, NCM 356, and CCD 841 CoN. As a comparative, we additionally analyzed the anti-inflammatory effects of Highbush blueberries on two inflamed colon cell cultures, NCM 356 and CCD 841 CoN. Furthermore, we sought to characterize the bioactive compounds in RCSC responsible for anti-cancer, anti-inflammatory, and immune-modulating effects. This work will be an essential starting point for understanding and identifying possible bioactive compounds in the fight against inflammatory bowel disease (IBD) and various cancers.

Fluorescent microscopy, flow cytometry, and quantitative polymerase chain reaction (qPCR) were performed, coupled with various bioassays, such as nuclear and cytoplasmic reactive oxygen species scavenging capabilities, propidium iodide cell viability assay, and gene microarrays to identify immunological gene targets. In doing so both RCSC and Highbush blueberry extracts were quantified for their anti-inflammatory and immune-modulating functions.

Finally, several organic solvents and techniques were utilized to identify possible bioactive compounds present in RCSC, Rotor evaporation, centri-vaped, column chromatography, thin layer chromatography (TLC), and high performance liquid chromatography (HPLC). Because of monetary and time restrictions we were unable toidentify the single responsible bioactive compound, if the case, however we were able to identify bioactive fractions consisting of eight compounds with anti-cancer and anti-inflammatory properties with great similarities to the whole RCSC. This will serve as a starting point for characterization of the bioactive compound(s)

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