Blog Post#3

Cryptosporidium parvum is an intracellular parasitic protozoan that causes cryptosporidiosis, a diarrheal disease occurring in humans and animals. Not only does C. parvum remain as one of the top organisms responsible for debilitating diarrheal infections in developing countries, it is also known to cause outbreaks in industrialized countries. The main way C. parvum spreads is via contaminated water. Due to their small size and high resistance to chlorination and other disinfectants, water filtration systems have had little success in the removal of the parasite from water sources and early detection measures are still being researched. Individuals with healthy immune systems who contract this disease will experience symptoms of unrelenting diarrhea, fever, dehydration, vomiting, and nausea, which may last up to two or three weeks. Treatment for these individuals involves drinking lots of fluids and rest. However, for immunocompromised individuals characterized by AIDS or cancer, the onset of the disease is relentless and very severe, often fatal. Currently, there is no effective treatment for cryptosporidiosis and a new chemotherapeutic drug is needed.

Previous research has shown that the polyamines: putrescine, spermidine and spermine, have all been found to be essential for the growth, development, and metabolism of C. parvum. Spermidine: Spermine N1- Acetyltransferase (SSAT) is a major regulator for polyamine synthesis, particularly spermine, and without it the parasite will surely die. As a result, last semester at Haskins Laboratories I focused on cloning and purifying the SSAT protein so that I could perform several assays on different substrates to develop a substrate curve. Due to Hurricane Sandy, my research was significantly affected and I had to restart the cloning and purification process. However, I have had a breakthrough and have successfully cloned and purified the SSAT protein needed to proceed. I am currently in the process of running multiple assays with substrates such as spermine, spermidine, histone, dapsone, and tris-HCl. The data obtained from these assays will generate substrate curves that will enable us to differentiate if SSAT is a general N1-acetyltransferase (GNAT) or if is a specific for spermine. If the GNAT is more active with other substrates, rather than with spermine, then it is probably not a true spermidine:spermine acetyltransferase. This is troublesome because we want to develop drugs that can inhibit the major polyamine spermine in order to inhibit the growth, development, and metabolism of C. parvum, thus potentially being able to cure cryptosporidiosis.

If these results indicate SSAT is truthfully the regulator of spermine then the process in synthesizing a drug with inhibitory abilities could be the sole determinant factor impeding a cure. With the assistance of Dr. Jamielee Rizzo and many more months of research our overall goal may come to fruition. Throughout the academic year, Dr. Nigel Yarlett has been an excellent mentor and role model. With his expertise and wisdom I have become fascinated with my research project and I have also learned to look past initial failures and reflect on my successes as a researcher.

 

—Emmanuel Bujans—
Pace University-Pforzheimer Honors College
Biochemistry Class of 2014
Research Assistant at Haskins Laboratories
(NERA) Medprep Scholar

Tel: (239) 961-9238
eb53946n@pace.edu

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