Cell and Developmental Biology Commons^™

Afm Study Of Structural Changes In Supported Planar Dppc Bilayers Containing General Anesthetic Isofluorane (Plus Additional Afm Experiments), Chad Mckell, Hiram Conley, David D. Busath

Faculty Publications

Atomic force microscopy (AFM) is a remarkable tool for assessing the structural properties of supported lipid planar bilayers under different physiological conditions. Previous work has shown that incorporation of anesthetics into artificial lipid bilayers results in domain formation [1], destruction of lipid aggregates and patches [2], anesthetic-lipid mixed micelle formation [2], and the development of interdigitated phases of reduced thickness compared to anesthetic-free bilayers [3]. In particular, these interdigitated phases are suspected to affect the structure and activity of membrane proteins, such as ion channels, and thus further research with proteinembedded bilayers exposed to anesthetics could reveal the mechanism responsible …

Real-Time Pcr Method For The Quantification Of Burkholderia Cepacia Complex Attached To Lung Epithelial Cells And Inhibitionn Of That Attachment, Ciara Wight, Gillian Herbert, Ruth Pilkington, Máire Callaghan, Siobhan Mcclean

Articles

To develop a rapid method to quantify the attachment of the cystic fibrosis pathogen, Burkholderia multivorans, to lung epithelial cells (16HBE14o(-)) using real-time PCR with a view to monitoring potential inhibition of lung cell attachment. Mammalian and bacterial DNA were purified from bacteria attached to lung epithelial cells. The relative amount of bacteria attached was determined by amplification of the recA gene relative to the human GAPDH gene, in the presence of SYBR Green. The method was thoroughly validated and shown to correlate well with traditional plating techniques. Inhibition of bacterial attachment with simple sugars was then evaluated by real-time …

Ornithine Decarboxylase (Odc) Regulation By Mtor: New Insights From Tsc1 Knockout Cells, Alyssa Mae Cates

Honors Program Theses

The polyamine pathway, when improperly regulated, is one central cause of skin carcinogenesis. The purpose of this project was to investigate the relationship between two biochemical signaling pathways, the mTOR (mammalian target of rapamycin) pathway and the polyamine pathway. It is known that the protein mTOR serves to regulate a key protein in the polyamine pathway, ornithine decarboxylase (ODC), but the precise mechanism remains unclear. It has been postulated that mTOR acts to regulate ODC through two RNA binding proteins, HuR (a member of the Hu family of proteins) and TIP (tristetraprolin). The project entailed characterizing that relationship using TSCl …

Adjacent Slice Prostate Cancer Prediction To Inform Maldi Imaging Biomarker Analysis, Shao-Hui Chuang, Xiaoyan Sun, Lisa Cazares, Julius Nyalwidhe, Dean Troyer, O. John Semmes, Jiang Li, Frederic D. Mckenzie, Nico Karssemeijer (Ed.), Ronald M. Summers (Ed.)

Electrical & Computer Engineering Faculty Publications

Prostate cancer is the second most common type of cancer among men in US [1]. Traditionally, prostate cancer diagnosis is made by the analysis of prostate-specific antigen (PSA) levels and histopathological images of biopsy samples under microscopes. Proteomic biomarkers can improve upon these methods. MALDI molecular spectra imaging is used to visualize protein/peptide concentrations across biopsy samples to search for biomarker candidates. Unfortunately, traditional processing methods require histopathological examination on one slice of a biopsy sample while the adjacent slice is subjected to the tissue destroying desorption and ionization processes of MALDI. The highest confidence tumor regions gained from the …

A Chemical Genetics Approach To Elucidate Mechanisms Of The Fission Yeast Polo Kinase In Cell Division, Jennifer Phelan, Dawn Clifford Hart

Student Summer Scholars Manuscripts

Cell division is a fundamental biological event that underlies the growth and development of all organisms. Because human and fission yeast (Schizosaccharomyces pombe) cells divide symmetrically through constriction of the actomyosin ring, fission yeast provides an ideal model system to reveal conserved cell cycle properties. In fission yeast, an evolutionarily conserved protein, Mid1, plays a critical role in organizing the early steps of contractile ring formation. Mid1 functions as a scaffold to bridge the cell cortex with the contractile ring. Cells lacking mid1 form off-centered, highly disorganized ring structures and exhibit severe cell division defects. Our previous research …

Chemosensitization Of Cancer Cells By Sirna Using Targeted Nanogel Delivery, Erin B. Dickerson, William H. Blackburn, Michael H. Smith, Laura B. Kapa, L. Andrew Lyon, John F. Mcdonald

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Background: Chemoresistance is a major obstacle in cancer treatment. Targeted therapies that enhance cancer cell sensitivity to chemotherapeutic agents have the potential to increase drug efficacy while reducing toxic effects on untargeted cells. Targeted cancer therapy by RNA interference (RNAi) is a relatively new approach that can be used to reversibly silence genes in vivo by selectively targeting genes such as the epidermal growth factor receptor (EGFR), which has been shown to increase the sensitivity of cancer cells to taxane chemotherapy. However, delivery represents the main hurdle for the broad development of RNAi therapeutics.

Methods: We report here …

Chapter 26 – Multiple Color Single Molecule Tirf Imaging And Tracking Of Maps And Motors, Jennifer L. Ross, Ram Dixit

Biology Faculty Research

Microtubules are part of a complex mechano-chemical network inside cells. In order to understand how the components of these systems work together, careful in vitro experiments must be performed with added complexity. These experiments can ideally image all the interacting species. In order to image these molecules, multiple-color fluorescence imaging can be performed. In this chapter, we describe some methods for performing multiple-color single molecule fluorescence imaging using total internal reflection fluorescence microscopy. We give several specific examples of species of microtubule-associate proteins and motors that can be examined with detailed protocols for labeling, purification, and imaging.

Chapter 27 – Studying Plus-End Tracking At Single Molecule Resolution Using Tirf Microscopy, Ram Dixit, Jennifer L. Ross

Biology Faculty Research

The highly dynamic microtubule plus-ends are key sites of regulation that impact the organization and function of the microtubule cytoskeleton. Much of this regulation is performed by the microtubule plus-end tracking (+TIP) family of proteins. +TIPs are a structurally diverse group of proteins that bind to and track with growing microtubule plus-ends in cells. +TIPs regulate microtubule dynamics as well as mediate interactions between microtubule tips and other cellular structures. Most +TIPs can directly bind to microtubules in vitro; however, the mechanisms for their plus-end specificity are not fully understood. Cellular studies of +TIP activity are complicated by the fact …

Mitochondrial Physiology Of Diapausing And Developing Embryos Of The Annual Killifish Austrofundulus Limnaeus: Implications For Extreme Anoxia Tolerance, Jeffrey M. Duerr, Jason E. Podrabsky

Biology Faculty Publications and Presentations

Diapausing embryos of the annual killifish Austrofundulus limnaeus have the highest reported anoxia tolerance of any vertebrate and previous studies indicate modified mitochondrial physiology likely supports anoxic metabolism. Functional mitochondria isolated from diapausing and developing embryos of the annual killifish exhibited VO2, respiratory control ratios (RCR), and P:O ratios consistent with those obtained from other ectothermic vertebrate species. Reduced oxygen consumption associated with dormancy in whole animal respiration rates are correlated with maximal respiration rates of mitochondria isolated from diapausing versus developing embryos. P:O ratios for developing embryos were similar to those obtained from adult liver, but were diminished in …