Biomedical Engineering and Bioengineering Commons^™

Population Fit Threshold: Fully Automated Signal Map Generation For Baseline Correction In Nmr-Based Metabolomics, Daniel C. Homer

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Baseline correction of NMR-based metabolomic spectra is a key step in data processing to elucidate biomarkers of diseased and toxic states. Automated baseline correction methods often use human-selected parameters. Presented is a method for automated baseline correction using parameters selected from and without manipulation of the spectrum. The focus is on generating an accurate signal map based on the differentiating characteristics of baseline noise and signal. The presented method of signal map generation and baseline correction was developed and tested on 13C and 1H NMR spectra. The spectra were as simple as chemical standards containing less than 25 signal peaks …

Charaterization Of Chlorpyrifos Toxicity On The Pancreatic Beta Cell Line Rinm5f, Zhongyu Yan

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Chlorpyrifos (CPF) is a well-known organophosphate (OP) insecticide that inhibits acetylcholinesterase (AChE). Organophosphates have been shown to induce hyperglycemia in both humans and animals that cannot be explained by AChE inhibition alone. Pancreatic β-cells are responsible for secreting insulin and playing a fundamental role in glucose homeostasis. The aim of this research was to investigate CPF toxicity on the insulin secreting β-cell line RINm5f (RIN). In the first part of this dissertation, we determined that cytotoxic concentrations of CPF between 50 µM and 200 µM induced a dose-dependent increase in RIN cell apoptotic and necrotic death (p<0.05). Lower doses, i.e., 5-25 µM did not induce apoptosis or necrosis. In further mechanistic studies, inhibitors of c-Jun N-terminal kinase (JNK) and P38 pathways, i.e., SP600125 and SB202190, successfully attenuated CPF-induced apoptosis (p<0.05), suggesting cell death is mediated by JNK and/or p38. In addition, N-Acetyl-L-cysteine (NAC), an antioxidant, inhibited apoptosis suggesting that oxidative stress is also involved in RIN cell death. In the second part of this dissertation, low dose CPF's effects on insulin synthesis and secretion was studied. Our results showed that RIN cells treated with CPF for 4 hr exhibited a dose-dependent decrease in insulin secretion in response to stimulation by 50 mM potassium (p<0.05). Insulin secretion stimulated by 25 mM glucose also showed significant decreases upon incubation with various doses of CPF (p<0.05). Insulin synthesis involves proteolytic cleavage of proinsulin at two specific sites. The enzymes responsible for promoting proinsulin cleavage, i.e, prohormone convertase (PC) 1/3 and 2 (PC2), contain a serine hydrolase catalytic triad, His-Asp-Ser, similar to AChE. CPF modification of PC enzymes similar to AChE, could potentially interrupt the conversion of proinsulin to insulin and disrupt insulin secretion. PC2 enzyme activity was significantly inhibited by CPF at concentrations as low as 10 µM (p<0.01). RIN cells, for the first time, were found to secrete proinsulin was through a constitutive instead of a regulated pathway. In summary, our results show that CPF not only exerts a direct cytotoxic effect at concentrations above 50 µM, but also significantly impairs insulin secretion at lower doses. Low dose CPF could potentially affect endocrine function in vivo by inhibiting the proinsulin processing enzyme PC2.

Extent And Effects Of Selection To Reduce Synthetic Cost Of Highly Expressed Proteins, Esley Marvin Heizer Jr

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Organisms that preferentially utilize less biosynthetically expensive amino acids in highly expressed genes exhibit metabolic efficiency. Exploration of this phenomenon has been limited to six prokaryotes.

I present a large scale analysis of metabolic efficiency in prokaryotic organisms and analysis of two eukaryotes (Saccharomyces cerevisiae and humans). Examination of 73 bacteria reveals the presence of metabolic efficiency in 66 organisms. The average correlation between amino acid biosynthetic cost and CAI scores in these organisms is -0.21. The seven organisms that did not exhibit metabolic efficiency are all Lactobacilli and highly auxotrophic. Saccharomyces cerevisiae exhibits significant trends between average amino acid …

Radio Frequency Controlled Stimulation Of Intracellular Gold Or Silver Nanoparticle Conjugates For Use As Potential Sensors Or Modulators Of Biological Function, Richard Craig Murdock

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The objective of this investigation was to further explore the ability of a radio frequency (RF) signal to interact with metal nanoparticles (NPs) for the purposes of remote stimulation or control of cellular processes. Gold and silver NPs have shown the ability to be heated by electromagnetic frequencies, especially for the purpose of deactivating abnormal cells. However, the heating of the NPs in these previous studies was constant and uncontrolled. For this study, our hypothesis was by investigating the RF heating characteristics of NPs and its impact on surface functionalizations, a remotely stimulated control mechanism can be developed at the …

Understanding The Molecular Dynamics Of Ypel3 And Fhit Gene Expression, Kevin Daniel Kelley

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A comprehensive understanding of the molecular signaling pathways that regulate cell growth and proliferation is essential in the realization of new therapeutic options to facilitate early detection and eradication of malignancy. Understanding the transcriptional regulation of the YPEL3 and FHIT genes forms the basis of this dissertation. YPEL3, or Yippee-like 3, is a newly identified p53 target gene that inhibits tumor cell growth and is thus itself, a novel tumor suppressor gene. FHIT, or Fragile histidine triad, is a well known tumor suppressor gene and is regulated at the transcriptional level by another growth inhibitory protein, FOXO3a, a Forkhead box …

Characterization And Improvement Of A Cone-Beam Ct Scanner For Quantitative Imaging, Jimish Dilip Joshi

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Computed tomography (CT) has various applications in different fields. In our case, the cone-beam CT scanner is used in the industrial field for qualitative and quantitative assessment of Procter and Gamble products. The Flash CT scanner, on which we do our research, has a microfocus x-ray tube, a detector size of 30 cm x 40 cm and a rotating stage for the object to be imaged. The x-ray tube can be operated up to 225 kV. Non-linearities in the response of the detector, scatter and beam hardening may cause false interpretations of the image values. To allow appropriate assessment of …

Cellular Effects Of Replicating A Polypurine-Polypyrimidine Sequence And The Interactions Of Due-B With Replication Proteins, Shere Lynne Myers

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This work investigates two questions regarding DNA replication. The first aim examines the interactions of DUE-B with replication proteins and the second explores the cellular effects of replicating a polypurine polypyrimidine sequence in human cells.

DUE-B siRNA decreases the chromatin binding of essential replication proteins Cdc45, PCNA and RPA. DUE-B also co-immunoprecipitates with Cdc45 and TopBP1. In vitro kinase assays suggest that the checkpoint protein ATR may phosphorylate DUE-B. These experiments lend further evidence that DUE-B plays an important role in the initiation of eukaryotic DNA replication.

To investigate the effects of replicating a polypurine-polypyrimidine sequence prone to secondary structure …

Induction Of P53 Dependent Cellular Senescence Through Hdmx Inhibition Or Ypel3 Expression, Kelly Lynn Robbins Miller

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Mutations in p53 that compromise its function have been reported in approximately half of all human cancers (Vousden et al., 2002). The other half of human tumors that retain wild-type p53 have a dysfunctional p53 pathway through other mechanisms (Wade et al., 2009). Activation of p53 leads to cell cycle arrest, DNA repair, apoptosis and senescence, however, the pathway leading to cellular senescence is the focus of this study. Cellular senescence is a process leading to irreversible arrest of cell division. Under normal physiologic conditions, the activity of p53 is kept in check by its negative regulators, Mdm2 and MdmX. …

Cytochrome C Oxidase From Rhodobacter Sphaeroides: Oligomeric Structure In The Phospholipid Bilayer And The Structural And Functional Effects Of A C-Terminal Truncation In Subunit Iii, Teresa L. Cvetkov

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Cytochrome c oxidase (COX) of the mitochondrial electron transport chain catalyzes the reduction of oxygen to water while concomitantly translocating protons across the inner mitochondrial membrane. This two part dissertation is a structural and functional investigation of COX using the bacterial model system from Rhodobacter sphaeroides (R.sph.).

First, the oligomeric structure of R.sph. COX within the lipid bilayer was investigated using discontinuous sucrose gradient ultracentrifugation. Utilizing this technique, liposomes containing R.sph. COX (pCOV) were separated from liposomes lacking enzyme (COV). The net buffering capacity and degree of light scattering per COX molecule were reduced in pCOVs, making them well suited …

Finding Cadaveric Human Head Masses And Center Of Gravity: A Comparison Of Direct Measurement To 3d Ing, Grant Corwin Roush

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Mass properties of the human head are critical elements in developing neck injury threshold criteria in acceleration and impact environments. In order to accurately simulate the dynamics of the head in impact and acceleration environments, valid mass properties data for the human head must exist.

The purpose of this study was two-fold: First, to directly measure and generate a useful data set of human head mass properties and anthropometry, and second, compare the results from the direct measurement to measurements obtained using computed tomographic (CT) analyses of the human head. Four cadaveric human heads, all male, were measured.

For the …

Computer-Based Skeletal Age Assessment Using Hand/Wrist Radiographs In Children 8-18 Years Old, Zhihong Ni

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Children's skeletons mature at different rates, and they can be affected by a variety of factors including disease, hormone imbalance or genetics. The assessment of skeletal maturity is a frequently performed procedure that allows the detection of hormonal, growth or genetic disorders. Several methods have been developed to estimate skeletal maturity. Most methods evaluate hand/wrist radiographs using indicators such as the ratios of various bone widths, the onset of the ossification of epiphysis and epiphyseal-diaphyseal fusion. Among those methods, the FELS method differs from others in the application of different grades to each indicator and the provision of a confidence …

A Biomechanical Comparison Of Locking Compression Plate Constructs With Plugs/Screws In Osteoporotic Bone Model, Krishna P. Desai

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Locking compression plates are proven to be safe for use in open reduction and internal fixation (ORIF) especially in osteoporotic bones. Because of various combinations of holes, the system provides more options for clinicians to use either locking screws or non-locking screws. This clinical research introduces screw like plugs which can be used along with the screws in case of locking compression plates. Experimental work was performed to determine the effectiveness of the plugs. The results showed that there is not a significant difference between the groups which used plugs and did not use the plugs, both in case of …