Biotechnology Commons^™

Comparison Of Subcellular Responses For The Evaluation And Prediction Of The Chemotherapeutic Response To Cisplatin In Lung Adenocarcinoma Using Raman Spectroscopy, Haq Nawaz, Franck Bonnier, Aidan Meade, Fiona Lyng, Hugh Byrne

Articles

Confocal Raman Micro spectroscopy (CRM) is employed to examine the chemical and physiological effects of anticancer agents, using cisplatin and A549 adenocarcinoma cells as a model compound and test system respectively. Spectral responses of the membrane and cytoplasm of the cell are analysed independently and the results are compared to previously reported spectroscopic studies of the nucleus. Moreover, Raman spectra from the proteins extracted from the control and exposed samples are acquired and analysed to confirm the origin of the molecular changes of the cell membrane and cytoplasm of the A549 cells. Multivariate data analysis techniques including Principal Component Analysis …

Copyright For Engineered Dna: An Idea Whose Time Has Come?, Christopher M. Holman

West Virginia Law Review

The rapidly emerging field of synthetic biology has tremendous potential to address some of the most compelling challenges facing our planet by providing clean renewable energy, nutri- tionally-enhanced and environmentally friendly agricultural products, and revolutionary new life-saving cures. However, leaders in the synthetic biology movement have voiced concern that biotechnology's current patent-centric approach to intellec- tual property is in many ways ill-suited to meet the challenge of synthetic biology, threatening to impede follow-on innovation and open access technology. For years, copyright and patent protection for computer software have existed side-by-side, the two forms of intellectual property complementing one another. Numerous …

Sherley V. Sebelius: Stem Cells And The Uneasy Interplay Between The Federal Bench And The Lab Bench, Ryan P. O'Quinn

Duke Law & Technology Review

After Barack Obama's election to the presidency, he promised that one of his top priorities in office would be to relieve the restrictions initiated by President George W. Bush on federal funding of embryonic stem cell research. President Obama followed through on his promise, but the celebrations in the nation's research labs were short-lived. Anti-abortion advocates and other scientists working in the field that would allegedly be out-competed in the federal funding arena brought a legal challenge to the new government position. The struggle culminated in August 2010 with a federal district court issuing a preliminary injunction to halt the …

Pcr Primers For The Detection Of Propane And Butane-Oxidizing Microorganisms, Brian Jeremy Chan

Master's Theses

In an increasingly energy-hungry world, our capacity to meet the heightened energy demands of the future has become a pressing matter. The most urgent of these concerns are tied to the accessibility of petroleum. Various experts have proselytized both the imminent arrival of peak oil production rates and the ensuing decline of those rates thereafter. And to that end, the development of novel and advanced oil exploration methodologies has become almost as important as finding the sources of oil themselves.

The soils above petroleum reservoirs play host to various communities of alkane- oxidizing bacteria that can utilize the natural gas …

Bacterial Killing By Dry Metallic Copper Surfaces, Christophe Espírito Santo, Ee Wen Lam, Christian Elowsky, Davide Quaranta, Dylan W. Domaille, Christopher J. Chang, Gregor Grass

Nebraska Center for Biotechnology: Faculty and Staff Publications

Metallic copper surfaces rapidly and efficiently kill bacteria. Cells exposed to copper surfaces accumulated large amounts of copper ions, and this copper uptake was faster from dry copper than from moist copper. Cells suffered extensive membrane damage within minutes of exposure to dry copper. Further, cells removed from copper showed loss of cell integrity. Acute contact with metallic copper surfaces did not result in increased mutation rates or DNA lesions. These findings are important first steps for revealing the molecular sensitive targets in cells lethally challenged by exposure to copper surfaces and provide a scientific explanation for the use of …

Fus Transgenic Rats Develop The Phenotypes Of Amyotrophic Lateral Sclerosis And Frontotemporal Lobar Degeneration, Cao Huang, Hongxia Zhou, Jianbin Tong, Han Chen, Yong-Jian Liu, Dian Wang, Xiaotao Wei, Xugang Xia

Nebraska Center for Biotechnology: Faculty and Staff Publications

Fused in Sarcoma (FUS) proteinopathy is a feature of frontotemporal lobar dementia (FTLD), and mutation of the fus gene segregates with FTLD and amyotrophic lateral sclerosis (ALS). To study the consequences of mutation in the fus gene, we created transgenic rats expressing the human fus gene with or without mutation. Overexpression of a mutant (R521C substitution), but not normal, human FUS induced progressive paralysis resembling ALS. Mutant FUS transgenic rats developed progressive paralysis secondary to degeneration of motor axons and displayed a substantial loss of neurons in the cortex and hippocampus. This neuronal loss was accompanied by ubiquitin aggregation and …

Biophysical Characterization Of The Membrane Binding Domain Of The Pro-Apoptotic Protein Bax, Pranav Garg

Electronic Theses and Dissertations

The BCL-2 family of proteins tightly regulates the delicate balance between life and death. The pore forming Bax is a pro-apoptotic member belonging to this protein family. At the onset of apoptosis, monomeric cytoplasmic Bax translocates to the outer mitochondrial membrane, forms oligomeric pores thereby letting mitochondrial cytochrome c enter the cytosol and initiate the apoptotic cascade. The C-terminal "helix 9" is thought to mediate the membrane binding of BAX. A 20-amino acid peptide corresponding to Bax C-terminus (VTIFVAGVLTASLTIWKKMG) and two mutants where the two lysines are replaced with Glu (charge reversal mutant, EE) or Leu (charge neutralization mutant, LL) …

Comparative Manufacture And Cell-Based Delivery Of Antiretroviral Nanoformulations, Shantanu Balkundi, Ari S. Nowacek, Ram S. Veerubhotla, Han Chen, Andrea Martinez-Skinner, Upal Roy, R. Lee Mosley, Georgette Kanmogne, Xinming Liu, Alexander V. Kabanov, Tatiana Bronich, Joellyn Mcmillan, Howard E. Gendelman

Nebraska Center for Biotechnology: Faculty and Staff Publications

Nanoformulations of crystalline indinavir, ritonavir, atazanavir, and efavirenz were manufactured by wet milling, homogenization or sonication with a variety of excipients. The chemical, biological, immune, virological, and toxicological properties of these formulations were compared using an established monocyte-derived macrophage scoring indicator system. Measurements of drug uptake, retention, release, and antiretroviral activity demonstrated differences amongst preparation methods. Interestingly, for drug cell targeting and antiretroviral responses the most significant difference among the particles was the drug itself. We posit that the choice of drug and formulation composition may ultimately affect clinical utility.

Quantitative Analysis Of Lignocellulosic Components Of Non-Treated And Steam Exploded Barley, Canola, Oat And Wheat Straw Using Fourier Transform Infrared Spectroscopy, P. K. Adapa, L. G. Schonenau, Thomas Canam, T. Dumonceaux

Faculty Research & Creative Activity

Rapid and cost effective quantification of lignocellulosic components (cellulose, hemicelluloses and lignin) of agricultural biomass (barley, canola, oat and wheat) is essential to determine the effect of various pre-treatments (such as steam explosion) on biomass used as feedstock for the biofuel industry. Fourier Transformed Infrared (FTIR) spectroscopy was considered as an option to achieve this objective. Regression equations having R2 values of 0.89, 0.99 and 0.98 were developed to predict the cellulose, hemicelluloses and lignin compounds of biomass, respectively. The average absolute difference in predicted and measured cellulose, hemicellulose and lignin in agricultural biomass was 7.5%, 2.5%, and 3.8%, respectively.

Quantitative Analysis Of Lignocellulosic Components Of Non-Treated And Steam Exploded Barley, Canola, Oat And Wheat Straw Using Fourier Transform Infrared Spectroscopy, P. Adapa, L. Schonenau, Thomas Canam, T. Dumonceaux

Faculty Research & Creative Activity

Rapid and cost effective quantification of lignocellulosic components (cellulose, hemicelluloses and lignin) of agricultural biomass (barley, canola, oat and wheat) is essential to determine the effect of various pre-treatments (such as steam explosion) on biomass used as feedstock for the biofuel industry. Fourier Transformed Infrared (FTIR) spectroscopy was considered as an option to achieve this objective. Regression equations having R2 values of 0.89, 0.99 and 0.98 were developed to predict the cellulose, hemicelluloses and lignin compounds of biomass, respectively. The average absolute difference in predicted and measured cellulose, hemicellulose and lignin in agricultural biomass was 7.5%, 2.5%, and 3.8%, respectively.

Chemical Characterization And In Vitro Fermentation Of Brassica Straw Treated With The Aerobic Fungus, Trametes Versicolor, J. E. Ramirez-Bribiesca, Y. Wang, L. Jin, Thomas Canam, J. R. Town, A. Tsang, T. J. Dumonceaux, T. A. Mcallister

Faculty Research & Creative Activity

Brassica napus straw (BNS) was either not treated or was treated with two strains of Trametes versicolor; 52J (wild type) or m4D (a cellobiose dehydrogenase-deficient mutant) with four treatments. Glucose was provided to encourage growth of the mutant strain. All treatments with T. versicolor decreased (P<0.05) neutral-detergent fibre and increased (P<0.05) protein and the concentration of lignin degradation products in straw. Ergosterol was highest (P<0.05) in straw treated with B-52J, suggesting it generated the most fungal biomass. Insoluble lignin was reduced (P<0.05) in straw treated with B-52J and B-m4D, but not with B-m4Dg. Mannose and xylose concentration were generally higher (P<0.05) in straw treated with fungi, whereas glucose and galactose were lower as compared with C-BNS. The four treatments above were subsequently assessed in rumen in vitro fermentations, along with BNS treated with 5 N NaOH. Concentrations of total volatile fatty acids after 24 and 48h were lower (P<0.05) in incubations that contained BNS treated with T. versicolor as compared with C-BNSor NaOH-treated BNS. Compared with C-BNS, in vitrodry matter disappearance and gas production were increased (P<0.05) by NaOH, but not by treatment with either strain of T. versicolor. Although treatment with T. versicolor did release more lignin degradation products, it did not appear to provide more degradable carbohydrate to in vitro rumen microbial populations, even when a mutant strain with compromised carbohydrate metabolism was utilized. Production of secondary compounds by the aerobic fungi may inhibit rumen microbial fermentation.

Linear Methods For Analysis And Quality Control Of Relative Expression Ratios From Quantitative Real-Time Polymerase Chain Reaction Experiments, Robert B. Page, Arnold J. Stromberg

Biology Faculty Publications

Relative expression quantitative real-time polymerase chain reaction (RT-qPCR) experiments are a common means of estimating transcript abundances across biological groups and experimental treatments. One of the most frequently used expression measures that results from such experiments is the relative expression ratio (R_E), which describes expression in experimental samples (i.e., RNA isolated from organisms, tissues, and/or cells that were exposed to one or more experimental or nonbaseline condition) in terms of fold change relative to calibrator samples (i.e., RNA isolated from organisms, tissues, and/or cells that were exposed to a control or baseline condition). Over the past decade, several …

Rotavirus Nsp4: Cell Type-Dependent Transport Kinetics To The Exofacial Plasma Membrane And Release From Intact Infected Cells, Thomas F. Gibbons, Stephen M. Storey, Cecelia V. Williams, Avery Mcintosh, Deanne M. Mitchel, Rebecca D. Parr, Friedhelm Schroeder, Judith M. Ball

Faculty Publications

Background

Rotavirus NSP4 localizes to multiple intracellular sites and is multifunctional, contributing to RV morphogenesis, replication and pathogenesis. One function of NSP4 is the induction of early secretory diarrhea by binding surface receptors to initiate signaling events. The aims of this study were to determine the transport kinetics of NSP4 to the exofacial plasma membrane (PM), the subsequent release from intact infected cells, and rebinding to naïve and/or neighboring cells in two cell types.

Methods

Transport kinetics was evaluated using surface-specific biotinylation/streptavidin pull-downs and exofacial exposure of NSP4 was confirmed by antibody binding to intact cells, and fluorescent resonant energy …

Chemical Characterization And In Vitro Fermentation Of Brassica Straw Treated With The Aerobic Fungus, Trametes Versicolor, J. Ramirez-Bribiesca, Y. Wang, L. Jin, Thomas Canam, J. Town, A. Tsang, T. Dumonceaux, T. Mcallister

Faculty Research & Creative Activity

Brassica napus straw (BNS) was either not treated or was treated with two strains of Trametes versicolor; 52J (wild type) or m4D (a cellobiose dehydrogenase-deficient mutant) with four treatments. Glucose was provided to encourage growth of the mutant strain. All treatments with T. versicolor decreased (P<0.05) neutral-detergent fibre and increased (P<0.05) protein and the concentration of lignin degradation products in straw. Ergosterol was highest (P<0.05) in straw treated with B-52J, suggesting it generated the most fungal biomass. Insoluble lignin was reduced (P<0.05) in straw treated with B-52J and B-m4D, but not with B-m4Dg. Mannose and xylose concentration were generally higher (P<0.05) in straw treated with fungi, whereas glucose and galactose were lower as compared with C-BNS. The four treatments above were subsequently assessed in rumen in vitro fermentations, along with BNS treated with 5 N NaOH. Concentrations of total volatile fatty acids after 24 and 48h were lower (P<0.05) in incubations that contained BNS treated with T. versicolor as compared with C-BNSor NaOH-treated BNS. Compared with C-BNS, in vitrodry matter disappearance and gas production were increased (P<0.05) by NaOH, but not by treatment with either strain of T. versicolor. Although treatment with T. versicolor did release more lignin degradation products, it did not appear to provide more degradable carbohydrate to in vitro rumen microbial populations, even when a mutant strain with compromised carbohydrate metabolism was utilized. Production of secondary compounds by the aerobic fungi may inhibit rumen microbial fermentation.

Biological Activity And Mass Spectrometric Analysis Of Vernonia Amygdalina Fractions, Broderick Eribo

Department of Biology Faculty Publications

Vernonia amygdalina (VA) is a tropical African plant of the Asteraceae family used as dietary supplement and as a medicinal herb. In this study, fatty acid esters, fatty acid amide, triterpene, diterpene alcohols and phytostanols were identified as the major chemical groups in the aqueous and organic fractions of VA leaf extracts. Their structures were elucidated, on the basis of GC-MS data, as hexanedioic acid, bis (2-ethylhexyl) ester (2a), erucamide (2b) and squalene (2c) in the water fraction; hexadecanoic acid methyl ester (3a), 9(Z)12(Z)15(Z)-octadecatrienoic acid, methyl ester (3b) and phytol (3c) in methanol fraction; phytol (4a), squalene (4b) and 7,22-ergostadienol …

Mne1 Is A Novel Component Of The Mitochondrial Splicing Apparatus Responsible For Processing Of A Cox1 Group I Intron In Yeast, Talina Watts, Oleh Khalimonchuk, Rachel Z. Wolf, Edward M. Turk, Georg Mohr, Dennis R. Winge

Department of Biochemistry: Faculty Publications

Saccharomyces cerevisiae cells lacking Mne1 are deficient in

intron splicing in the gene encoding the Cox1 subunit of cytochrome

oxidase but contain wild-type levels of the bc1 complex.

Thus, Mne1 has no role in splicing of COB introns or expression

of the COB gene. Northern experiments suggest that splicing of

the COX1 aI5β intron is dependent on Mne1 in addition to the

previously known Mrs1, Mss116, Pet54, and Suv3 factors. Processing

of the aI5_ intron is similarly impaired in mne1∆ and

mrs1∆ cells and overexpression of Mrs1 partially restores the

respiratory function of mne1∆ cells. Mrs1 …

Efficacy Of The Pentavalent Rotavirus Vaccine, Rotateq (Rv5), Between Doses Of A 3-Dose Series And With Less Than 3 Doses (Incomplete Regimen), Penelope D. Dennehy, Timo Vesikari, David O. Matson, Robbin F. Itzler, Michael J. Dallas, Michelle G. Goveia, Mark J. Dinubile, Penny M. Heaton, Max Ciarlet

Community & Environmental Health Faculty Publications

Post-hoc analyses of the Rotavirus Efficacy and Safety Trial (RES T) were conducted to determine whether the pentavalent rotavirus vaccine (RV5) confers early protection against rotavirus gastroenteritis (RVGE) before completion of the 3-dose regimen. To evaluate the efficacy of RV5 between doses in reducing the rates of RVGE-related hospitalizations and emergency department (ED) visits in infants who ultimately received all 3 doses of RV5/placebo, events occurring from 2 weeks after the first and second doses to receipt of the subsequent dose (Analysis A) and events occurring from 2 weeks after the first and second doses to 2 weeks after the …

Reduced Utilization Of Selenium By Naked Mole Rats Due To A Specific Defect In Gpx1 Expression, Marina V. Kasaikina, Alexei V. Lobanov, Mikalai I. Malinouski, Byung Cheon Lee, Javier Seravalli, Dmitri E. Fomenko, Anton A. Turanov, Lydia Finney, Stefan Vogt, Thomas J. Park, Richard A. Miller, Dolph L. Hatfield, Vadim N. Gladyshev

Department of Biochemistry: Faculty Publications

Naked mole rat (MR) Heterocephalus glaber is a rodent model of delayed aging because of its unusually long life span (>28 years). It is also not known to develop cancer. In the current work, tissue imaging by x-ray fluorescence microscopy and direct analyses of trace elements revealed low levels of selenium in the MR liver and kidney, whereas MR and mouse brains had similar selenium levels. This effect was not explained by uniform selenium deficiency because methionine sulfoxide reductase activities were similar in mice and MR. However, glutathione peroxidase activity was an order of magnitude lower inMRliver and kidney …

Defining A Relationship Between Dietary Fatty Acids And The Cytochrome P450 System In A Mouse Model Of Fatty Liver Disease, Monika Gonzalez, Whitney Sealls, Elliot D. Jesch, M. Julia Brosnan, Istvan Ladunga, Xinxin Ding, Paul N. Black, Concetta C. Dirusso

Department of Biochemistry: Faculty Publications

Liver-specific ablation of cytochrome P450 reductase in mice (LCN) results in hepatic steatosis that can progress to steatohepatitis characterized by inflammation and fibrosis. The specific cause of the fatty liver phenotype is poorly understood but is hypothesized to result from elevated expression of genes encoding fatty acid synthetic genes. Since expression of these genes is known to be suppressed by polyunsaturated fatty acids, we performed physiological and genomics studies to evaluate the effects of dietary linoleic and linolenic fatty acids (PUFA) or arachidonic and decosahexaenoic acids (HUFA) on the hepatic phenotypes of control and LCN mice by comparison with a …

Functional Diversification Of Thylakoidal Processing Peptidases In Arabidopsis Thaliana, Shih-Chi Hsu, Joshua K. Endow, Nicholas J. Ruppel, Rebecca Roston, Amy J. Baldwin, Kentaro Inoue

Department of Biochemistry: Faculty Publications

Thylakoidal processing peptidase (TPP) is responsible for removing amino-terminal thylakoid-transfer signals from several proteins in the thylakoid lumen. Three TPP isoforms are encoded by the nuclear genome of Arabidopsis thaliana. Previous studies showed that one of them termed plastidic type I signal peptidase 1 (Plsp1) was necessary for processing three thylakoidal proteins and one protein in the chloroplast envelope in vivo. The lack of Plsp1 resulted in seedling lethality, apparently due to disruption of proper thylakoid development. The physiological roles of the other two TPP homologs remain unknown. Here we show that the three A. thaliana TPP isoforms …