- ItemOpen AccessPurification of phenolic glycolipid from mycobacterium tuberculosis(Colorado State University. Libraries, 2009) Vigil, Scott Brandon, author; Hesser, Danny C., author; Dobos, Karen, authorMycobacterium tuberculosis (Mtb) is a very successful human pathogen, infecting one-third of the world's population. Although originally thought to have little to no genetic variation, Mtb has been recently shown to be distinguished to clades based on geographical distribution. Further, members of specific clades (and/or subclades) have been shown to possess differences in virulence. Several clades from Asia, Africa, and India were found to express phenolic glycolipid (PGL). PGL is absent in many North American and European clades due to gene deletion. The West Beijing strains of Mtb, HN878, W4, and W451, contain PGL as part of their cellular envelope. These strains demonstrated reduced production of important immune-mediating cytokines including tumor necrosis factor alpha and interleukin 12. Subsequent evidence demonstrated this was due to PGL. Thus, it is believed that PGL works in concert with other factors to enhance the virulence of Mtb. Purification of PGL from these strains will permit further studies on the molecular interactions of PGL during infection with Mtb. In our laboratory, we attempted to isolate PGL from Mtb strain HN878. Initially, a lipid extraction was performed with 1:2 chloroform: methanol. This extract was then analyzed via preparatory and analytical Thin-Layer Chromatography (TLC) plates with a 90:10 and 95:5 chloroform: methanol developing solution. These TLC plates were then sprayed with CuSO4 and α-naphthol for sugar and carbon compound detection. Ultra-violet light was used to view and outline what is believed to be the major PGL band. This band was scraped off the TLC and will be extracted with diethyl ether to purify PGL. Once purified, we will perform studies with naïve and Mtb-infected macrophages to assess the role of PGL in the pro-inflammatory response during infection.
- ItemOpen AccessDoes Flotillin play a role in lipid raft organization of the GnRH receptor and its ability to transduce an intracellular signal?(Colorado State University. Libraries, 2009) Phillips, Adam Paul, author; Mrdutt, Mary Megan, author; Clay, Colin McKeown, authorGonadotropin-Releasing Hormone (GnRH) and its subsequent signaling through the GnRH Receptor (GnRH-R) is critical for gonadal development and control of reproduction function. The GnRH-R is a member of the G-protein coupled receptor (GPCR) superfamily and is localized to specialized low-density areas on the cell membrane termed lipid rafts. These raft domains are implicated in GPCR coupled signaling by spatially organizing receptors and their associated signaling proteins to specific domains in the plasma membranes of cells. These raft domains appear to play an important role in the organization of GnRH-R and the signaling of GnRH to MAPkinase. Flotillin-1 is a protein thought to be intricately involved in the organization of rafts and the trafficking of proteins to raft domains. To examine the potential role of flotillin in GnRH signaling, gonadotrope derived αT3-1 cells were transfected with a specific siRNA for Flotillin-1 with the long-term goal of assessing the impact of Flotillin-1 deficiency on GnRHR trafficking to lipid rafts and signaling to intracellular targets including extracellular signal regulated kinase (ERK). Objectives: to use si-GLO Red to determine the transfection efficiency of Flotillin-1 siRNA; to use siRNA technology to knockdown Flotillin-1 expression; to determine the effect of Flotillin-1 knockdown on GnRH-R signaling.
- ItemOpen AccessThe relationship between cognitive inhibition and extraversion/introversion(Colorado State University. Libraries, 2009) Knapp, Alexandra Elizabeth, author; Campbell, Alana May, author; Burr, Hilary Kay, author; Matigian, Morgan Ashley, author; Davalos, Deana, authorThe purpose of this study was to analyze the visual and textual content presented on the covers of Seventeen magazines published between 1997 and 2007. Seventeen is the most widely read magazine among adolescent females today (SRDS, 2002); research suggests that young readers look to this publication for ideas about who to be and how to look (Duffy and Gotcher, 1996). Covers were chosen for analysis because they represent an index to the information included within the magazine and serve as an advertisement for the sale of the publication. Since young people look to Seventeen magazine for insight on how to look and act, it is important to be aware of what they are being told and shown.
- ItemOpen AccessThe role of the human Translationally Controlled Tumor Protein (TCTP) in mitosis(Colorado State University. Libraries, 2009) Buechler, Betsy Lauren, author; Deluca, Jennifer G., authorMitosis is the complex process that results in division of DNA and other cellular components into two daughter cells. Successful cell division requires that microtubules of the mitotic spindle attach to the kinetochores of mitotic chromosomes. These attachments are used for both aligning chromosomes at the spindle equator and for the physical separation of sister chromosomes during anaphase. In the study of how microtubules and kinetochores work together during mitosis, researchers are searching for new proteins that may help to piece together this complex puzzle. The Translationally Controlled Tumor Protein (TCTP) has been suggested to play a role in mitotic cell division, as it has been immuno-localized to the mitotic spindle, however, its precise function in mitosis remains unknown. The goal of my project is to determine the role of human TCTP in mitosis. Thus far, my research has shown that TCTP is located at the microtubules during mitosis, and when knocked down by siRNA treatment, human HeLa H2B-GFP cells (GFP-tagged chromosomes) are either unable to complete mitosis or take an extended time to do so. Currently, I am undergoing procedures to pick the most suitable human cell line in which to study the roles of TCTP during mitosis (HeLa-S3, HeLa-Kyoto, or HeLa H2B-GFP), and a mitotic index and characterization of this cell line. In the future, I plan to look in to the association between F-actin and TCTP, recently brought to light by Bazile et al, 2009.