Research Methods in Life Sciences Commons^™

Itraq-Based Proteomics Analysis And Network Integration For Kernel Tissue Development In Maize, Long Zhang, Yongbin Dong, Qilei Wang, Chunguang Du, Wenwei Xiong, Xinyu Li, Sailan Zhu, Yuling Li

Department of Biology Faculty Scholarship and Creative Works

Grain weight is one of the most important yield components and a developmentally complex structure comprised of two major compartments (endosperm and pericarp) in maize (Zea mays L.), however, very little is known concerning the coordinated accumulation of the numerous proteins involved. Herein, we used isobaric tags for relative and absolute quantitation (iTRAQ)-based comparative proteomic method to analyze the characteristics of dynamic proteomics for endosperm and pericarp during grain development. Totally, 9539 proteins were identified for both components at four development stages, among which 1401 proteins were non-redundant, 232 proteins were specific in pericarp and 153 proteins were specific in …

Effects Of Food Consumption On Cell Proliferation In The Brain Of Python Regius, Stacy Star Habroun

Master's Theses

Neurogenesis is an important and vastly under-explored area in reptiles. While the ability to generate new brain cells in the adult mammalian brain is limited, reptiles are able to regenerate large populations of neuronal cells. Pythons exhibit a characteristic specific dynamic action (SDA) response after food intake with an increase in metabolic rate that facilitates processing the meal. Associated with this change in SDA, pythons (Python spp.) also exhibit impressive plasticity in their digestive and cardiovascular physiology due to the sheer magnitude of the increase in organ growth that occurs after a meal to speed digestion, absorption, and assimilation of …

Responsible Translation Of Stem Cell Research: An Assessment Of Clinical Trial Registration And Publications, Moses Fung, Yan Yuan, Harold Atkins, Qian Shi, Tania M. Bubela

Office of the Provost

We assessed the extent to which the publication of clinical trial results of innovative cell-based interventions reflects International Society for Stem Cell Research best practice guidelines. We assessed: (1) characteristics and time to publication of completed trials; (2) quality of reported trials; and (3) results of published trials. We identified and analyzed publications from 1,052 novel stem cell clinical trials: 179 (45.4%) of 393 completed trials had published results; 48 trials were registered by known stem cell tourism clinics, none of which reported results. Completed non-industry-sponsored trials initially published more rapidly, but differences with industry-sponsored trials decreased over time. Most …

Estimating Seasonal Variation In Condition Of Yellow Perch, Emily Anne Powers

Biology Graduate Theses

Research has shown that Yellow Perch Perca flavescens, found throughout lakes and streams of North America, are good indicators of environmental health. Due to the correlation between fish condition and ecosystem health it is essential to reliably estimate condition to better manage our fisheries. Therefore, the study objectives were to measure the seasonal changes in adult Yellow Perch percent dry weight (PDW), and compare the reliability of bioelectrical impedance analysis (BIA) and morphometric-based estimates of condition. The seasonal fluctuations in average monthly PDW were measured over a two- year period and were sinusoidal in shape with peaks in August 2015 …

Evolution Of Equine Arteritis Virus During Persistent Infection In The Reproductive Tract Of The Stallion And The Male Donkey, Bora Nam

Theses and Dissertations--Veterinary Science

Equine arteritis virus (EAV) establishes persistent infection in the stallion reproductive tract, and the carrier stallion continues to shed virus in semen for weeks to years or lifelong. The objective of this study was to elucidate the intra-host evolution of EAV during persistent infection in stallions. Seven EAV seronegative stallions were experimentally infected with EAV KY84 strain and followed for 726 days post-infection, and sequential clinical samples including semen were collected for virus isolation and next-generation sequencing (NGS). In addition, archived sequential semen samples from two stallions that were naturally infected with EAV KY84 for a long-period (up to 10 …

Guidelines To Avoid Typical Difficulties According To The Rubric For Experimental Design (Red), Annwesa Dasgupta, Nancy Pelaez

PIBERG Publications

Experimental design is an important component of undergraduate biology education as it generates knowledge of biology. Despite its importance, there is limited information about what students actually learn from designing experiments. Dasgupta et al (2014) reported on the development and validation of a Rubric for Experimental Design (RED), informed by a literature review and empirical analysis of thousands of undergraduate biology students’ responses to three published assessments. The RED is a useful probe for five major areas of experimental design abilities: the variable properties of an experimental subject; the manipulated variables; measurement of outcomes; accounting for variability; and the scope …

Alzheimer Disease Research In The 21st Century: Past And Current Failures, New Perspectives And Funding Priorities, Francesca Pistollato, Elan L. Ohayon, Ann Lam, Gillian R. Langley, Thomas J. Novak, David Pamies, George Perry, Eugenia Trushina, Robin S.B. Williams, Alex E. Roher, Thomas Hartung, Stevan Harnad, Neal D. Barnard, Martha Clare Morris, Mei-Chun Lai, Ryan Merkley, P. Charukeshi Chandrasekera

Experimentation Collection

Much of Alzheimer disease (AD) research has been traditionally based on the use of animals, which have been extensively applied in an effort to both improve our understanding of the pathophysiological mechanisms of the disease and to test novel therapeutic approaches. However, decades of such research have not effectively translated into substantial therapeutic success for human patients. Here we critically discuss these issues in order to determine how existing human-based methods can be applied to study AD pathology and develop novel therapeutics. These methods, which include patient-derived cells, computational analysis and models, together with large-scale epidemiological studies represent novel and …

Privatization Of Biomedical Research: A Short Term Answer With Long Term Consequences, Frederick Azalekor

Nebraska College Preparatory Academy: Senior Capstone Projects

Biomedical research owned by the government is now available to be owned by private institutions. This results in stagnant innovations because researchers have trouble with all the restrictions that are placed by private institutions. Privatization of biomedical research has short term positive effects but its negative effects are overarching and disastrous. One way to combat this, is for licenses to be deployed slowly with future consequences in mind.

• Privatization has short term benefits but long term disadvantages are disastrous. • To reduce damaging effect, privatization must be deployed slowly and carefully. • Patents or proprietary ownership must be given …

Lessons From Toxicology: Developing A 21st‑Century Paradigm For Medical Research, Gill Langley, Christopher P. Austin, Anil K. Balapure, Linda S. Birnbaum, John R. Bucher, Julia Fentem, Suzanne C. Fitzpatrick, John R. Fowle Iii, Robert J. Kavlock, Hiroaki Kitano, Brett A. Lidbury, Alysson R. Muotri, Shuang-Qing Peng, Dmitry Sakharov, Troy Seidle, Thales Trez, Alexander Tonevitsky, Anja Van De Stolpe, Maurice Whelan, Catherine Willett

Experimentation Collection

Biomedical developments in the 21st century provide an unprecedented opportunity to gain a dynamic systems-level and human-specific understanding of the causes and pathophysiologies of disease. This understanding is a vital need, in view of continuing failures in health research, drug discovery, and clinical translation. The full potential of advanced approaches may not be achieved within a 20th-century conceptual framework dominated by animal models. Novel technologies are being integrated into environmental health research and are also applicable to disease research, but these advances need a new medical research and drug discovery paradigm to gain maximal benefits. We suggest a new conceptual …

The Multifactorial Role Of The 3rs In Shifting The Harm-Benefit Analysis In Animal Models Of Disease, Melanie L. Graham, Mark J. Prescott

Biomedical Research and Alternative Methods Collection

Ethics on animal use in science in Western society is based on utilitarianism, weighing the harms and benefits to the animals involved against those of the intended human beneficiaries. The 3Rs concept (Replacement, Reduction, Refinement) is both a robust framework for minimizing animal use and suffering (addressing the harms to animals) and a means of supporting high quality science and translation (addressing the benefits). The ambiguity of basic research performed early in the research continuum can sometimes make harm-benefit analysis more difficult since anticipated benefit is often an incremental contribution to a field of knowledge. On the other hand, benefit …

Age-Dependent Breakdown Of The Blood Brain Barrier And Associated Changes In S100b Ko Mice, Eric V. Brown

Graduate School of Biomedical Sciences Theses and Dissertations

Autoantibodies play an important role in many autoimmune diseases. Recent research has shown that breakdown of the blood brain barrier (BBB) occurs concomitant to generation of brain reactive autoantibodies in many neurodegenerative diseases, which serve as biomarkers and drivers of pathology. SI00B, a calcium binding protein found most highly expressed in astrocytes which ensheathe the BBB, has many functions in neural development and signaling. Currently literature indicates that S100B KO mice develop normally, with no phenotypic abnormalities. Here, it is demonstrated that S100B KO mice seem to develop a chronic BBB breakdown similar to that seen in human neurodegenerative diseases. …

Evolution Of Sperm Activation In Nematodes, Qing Wei

Graduate School of Biomedical Sciences Theses and Dissertations

Three hermaphroditic species in the nematode genus Caenorhabditis - C. elegans, C. briggsae and C. tropicalis evolved independently from a male/female ancestor. Thus, the hermaphroditic reproductive system provides an ideal subject for studying the origin of new complex traits. Previous studies implied that this transition required two steps: a mutation that caused XY animals to make sperm, and a mutation that allowed XX spermatids to activate and fertilize oocytes. In this dissertation, I show how sperm activation is regulated in different hermaphroditic species.

First, I used TALENs to create genetic tools for C. briggsae and C. tropicalis. These tools were …

Genome Engineering To Create Dominant Alleles In Caenorhabditis Elegans Using Crispr-Cas9 Technology, Abrar Sulaimani

Graduate School of Biomedical Sciences Theses and Dissertations

Many investigators have being using CRISPR-Cas9 as a method of genome engineering because it is easy, accurate and fast. This technique has been used to modify the genomes of a wide variety of organisms, including the nematode Caenorhabditis elegans (C. elegans). The short life cycle and ease of introducing exogenous plasmids make C. elegans an ideal system for advancing this technique. My thesis had two aims that focused on developing methods to create dominant alleles in C. elegans. Genetic modifications like precise deletion and insertions into a locus of chromosome are technically challenging. Additionally, although there are several ways of …

Validating A Method For Measuring Protein Domain And Lipid Binding Interactions Using Luciferase Fusion Proteins, Dezaray E. Thoen

Honors Capstones

Capstone submitted as a graduation requirement for the BSU Honors Program.

Considering A New Paradigm For Alzheimer’S Disease Research, Gillian R. Langley

Experimentation Collection

Using Alzheimer’s disease as a case study, this review argues that it might be time to consider a new paradigm in medical research and drug discovery. The existing framework is overly dependent on often unvalidated animal models, particularly transgenic mice. Translational success remains elusive and costly late-stage drug failure is common. The conventional paradigm tends to overlook species differences and assumes that animal-based findings are generally applicable to humans. Could pathways-based research using advanced human-specific models probed with new tools, including those of systems biology, take centre stage? The current transition in chemical toxicology to a 21st-century paradigm could be …

Technique For Staining The Myelin-Like Sheath In Earthworms, Kayla Brown

Honors Capstones

Capstone submitted as a graduation requirement for the BSU Honors Program.

Moving Beyond The Welfare Standard Of Psychological Well-Being For Nonhuman Primates: The Case Of Chimpanzees, John P. Gluck

Experimentation Collection

Since 1985, the US Animal Welfare Act and Public Health Service policy have required that researchers using nonhuman primates in biomedical and behavioral research develop a plan ‘‘for a physical environment adequate to promote the psychological well-being of primates.’’ In pursuing this charge, housing attributes such as social companionship, opportunities to express species-typical behavior, suitable space for expanded locomotor activity, and nonstressful relationships with laboratory personnel are dimensions that have dominated the discussion. Regulators were careful not to direct a specific set of prescriptions (i.e., engineering standards) for the attainment of these goals, but to leave the design of the …

State-Of-The-Art Of 3d Cultures (Organs-On-A-Chip) In Safety Testing And Pathophysiology, Natalie Alépée, Anthony Bahinski, Mardas Daneshian, Bart De Wever, Alan Goldberg, Jan Hansmann, Thomas Hartung, John Haycock, Helena T. Hogberg, Lisa Hoelting, Jens M. Kelm, Suzanne Kadereit, Emily Mcvey, Robert Landsiedel, Marcel Leist, Marc Lübberstedt, Fozia Noor, Christian Pellevoisin, Dirk Petersohn, Uwe Pfannenbecker, Kerstin Reisinger, Tzutzuy Ramirez, Barbara Rothen-Rutishauser, Monika Schäfer-Korting, Katrin Zeilinger, Marie-Gabriele Zurich

in Vitro Research Models Collection

Integrated approaches using different in vitro methods in combination with bioinformatics can (i) increase the success rate and speed of drug development; (ii) improve the accuracy of toxicological risk assessment; and (iii) increase our understanding of disease. Three-dimensional (3D) cell culture models are important building blocks of this strategy which has emerged during the last years. The majority of these models are organotypic, i.e., they aim to reproduce major functions of an organ or organ system. This implies in many cases that more than one cell type forms the 3D structure, and often matrix elements play an important role. This …

Consensus Report On The Future Of Animal-Free Systemic Toxicity Testing, Marcel Leist, Nina Hasiwa, Costanza Rovida, Mardas Daneshian, David Basketter, Ian Kimber, Harvey Clewell, Tilman Gocht, Alan Goldberg, Francois Busquet, Anna-Maria Rossi, Michael Schwarz, Martin Stephens, Rob Taalman, Thomas B. Knudsen, James Mckim, Georgina Harris, David Pamies, Thomas Hartung

Experimentation Collection

Since March 2013, animal use for cosmetics testing for the European market has been banned. This requires a renewed view on risk assessment in this field. However, in other fields as well, traditional animal experimentation does not always satisfy requirements in safety testing, as the need for human-relevant information is ever increasing. A general strategy for animal-free test approaches was outlined by the US National Research Council’s vision document for Toxicity Testing in the 21st Century in 2007. It is now possible to provide a more defined roadmap on how to implement this vision for the four principal areas of …

Changing Body Condition In Walleye With Reference To Ideal Sample Size Requirements, Kevin Longtin

Journal of Earth and Life Science

The objective of this study was to determine if walleye Sander vitreus percent dry weight, a measure of condition, differed among summer months of 2014 on Maple Lake, Minnesota. A second objective was to determine if there was a relationship between condition and body temperature, total length, or wet weight. Between three and six walleyes were collected per month by hook and line methods. All fish had internal body temperatures taken at the time of capture and were oven dried in the laboratory at 80 °C in order to measure percent dry weight. There was no relationship between condition and …

Observing The Unwatchable Through Acceleration Logging Of Animal Behavior, Danielle D. Brown, Roland Kays, Martin Wikelski, Rory Wilson, A. Peter Klimley

Methodology and Animal Models in Research

Behavior is an important mechanism of evolution and it is paid for through energy expenditure. Nevertheless, field biologists can rarely observe animals for more than a fraction of their daily activities and attempts to quantify behavior for modeling ecological processes often exclude cryptic yet important behavioral events. Over the past few years, an explosion of research on remote monitoring of animal behavior using acceleration sensors has smashed the decades-old limits of observational studies. Animal-attached accelerometers measure the change in velocity of the body over time and can quantify fine-scale movements and body postures unlimited by visibility, observer bias, or the …

Incorporating New Technologies Into Toxicity Testing And Risk Assessment: Moving From 21st Century Vision To A Data-Driven Framework, Russell S. Thomas, Martin A. Philbert, Scott Auerbach, Barbara A. Wetmore, Michael J. Devito, Ila Cote, J. Craig Rowlands, Maurice Whelan, Sean M. Hays, Melvin E. Andersen, M.E. (Bette) Meek, Lawrence W. Reiter, Jason C. Lambert, Harvey J. Clewell Iii, Martin L. Stephens, Q. Jay Zhao, Scott C. Wesselkamper, Lynn Flowers, Edward W. Carney, Tim P. Pastoor, Dan D. Petersen, Carole L. Yauk, Andy Nong

in Vitro Research Models Collection

Based on existing data and previous work, a series of studies is proposed as a basis toward a pragmatic early step in transforming toxicity testing. These studies were assembled into a data-driven framework that invokes successive tiers of testing with margin of exposure (MOE) as the primary metric. The first tier of the framework integrates data from high-throughput in vitro assays, in vitro-to-in vivo extrapolation (IVIVE) pharmacokinetic modeling, and exposure modeling. The in vitro assays are used to separate chemicals based on their relative selectivity in interacting with biological targets and identify the concentration at which these interactions …

Modulation Of Bax/Bak Dependent Apoptosis By Sirtuin 3 And Mitochondrial Permeability Transition By Sirtuin 4, Manish Verma

Graduate School of Biomedical Sciences Theses and Dissertations

Mitochondria are dynamic organelles that regulate a myriad of cellular functions, including energy production and metabolic regulation. Mitochondria are also a critical regulator of cell death signaling cascades modulating both apoptotic and necrotic cell death. However, what determines which cell death pathway is activated is still unclear. The mitochondrial/intrinsic pathway of apoptosis is dependent on the activation of pro-apoptotic proteins, Bax and Bak, which induce mitochondrial outer membrane permeabilization (MOMP). Once the integrity of outer mitochondrial membrane (OMM) is compromised, pro-apoptotic intermembrane space proteins like cytochrome c, Smac/Diablo, Omi/HtrA2 and AIF are released into the cytoplasm, which activates the post-mitochondrial …

Can Acute Dermal Systemic Toxicity Tests Be Replaced With Oral Tests? A Comparison Of Route-Specific Systemic Toxicity And Hazard Classifications Under The Globally Harmonized System Of Classification And Labelling Of Chemicals (Ghs), Nigel P. Moore, David J. Andrew, Donald L. Bjerke, Stuart Creton, David Dreher, Thomas Holmes, Pilar Prieto, Troy Seidle, Tim G. Rowan

Experimentation Collection

Acute systemic toxicity data (LD50 values) and hazard classifications derived in the rat following oral administration and dermal application have been analysed to examine whether or not orally-derived hazard classification or LD50 values can be used to determine dermal hazard classification. Comparing the oral and dermal classifications for 335 substances derived from oral and dermal LD50 values respectively revealed 17% concordance, and indicated that 7% of substances would be classified less severely while 76% would be classified more severely if oral classifications were applied directly to the dermal route. In contrast, applying the oral LD50 values within the dermal classification …

The Use Of Biomarkers Of Toxicity For Integrating In Vitro Hazard Estimates Into Risk Assessment For Humans, Bas Blaauboer, Kim Boekelheide, Harvey Clewell, Mardas Daneshian, Milou M.L. Dingemans, Alan M. Goldberg, Marjoke Heneweer, Joanna Jaworska, Nynke I. Kramer, Marcel Leist, Hasso Seibert, Emanuela Testai, Rob J. Vandebriel, James D. Yager, Joanne Zurlo

in Vitro Research Models Collection

The role that in vitro systems can play in toxicological risk assessment is determined by the appropriateness of the chosen methods, with respect to the way in which in vitro data can be extrapolated to the in vivo situation. This report presents the results of a workshop aimed at better defining the use of in vitro-derived biomarkers of toxicity (BoT) and determining the place these data can have in human risk assessment. As a result, a conceptual framework is presented for the incorporation of in vitro-derived toxicity data into the risk assessment process. The selection of BoT takes into account …

Alternative Approaches For Medical Countermeasures To Biological And Chemical Terrorism And Warfare, Thomas Hartung, Joanne Zurlo

Biomedical Research and Alternative Methods Collection

The desire to develop and evaluate drugs as potential countermeasures for biological and chemical threats requires test systems that can also substitute for the clinical trials normally crucial for drug development. Animal models have limited predictivity for drug efficacy, as is well known from many disappointments in clinical trials. Traditional in vitro and in silico approaches are not really game changers here, but the substantial investment into novel tools now underway might bring about a second generation of alternative approaches. The avenue pursued focuses primarily on the development of a Human on a Chip, i.e., the combination of different three-dimensional …

Superglue Is Not Super: An Assessment Of Superglue For Suturing Tag Incisions In A Cultured Marine Fish, Vincent Raoult, Culum Brown, Jane E. Williamson

Aquaculture Collection

No abstract provided.

Accelerating The Development Of 21st-Century Toxicology: Outcome Of A Human Toxicology Project Consortium Workshop, Martin L. Stephens, Craig Barrow, Melvin E. Andersen, Kim Boekelheide, Paul L. Carmichael, Michael P. Holsapple, Mark Lafranconi

in Vitro Research Models Collection

The U.S. National Research Council (NRC) report on “Toxicity Testing in the 21st century” calls for a fundamental shift in the way that chemicals are tested for human health effects and evaluated in risk assessments. The new approach would move toward in vitro methods, typically using human cells in a high-throughput context. The in vitro methods would be designed to detect significant perturbations to “toxicity pathways,” i.e., key biological pathways that, when sufficiently perturbed, lead to adverse health outcomes. To explore progress on the report’s implementation, the Human Toxicology Project Consortium hosted a workshop on 9–10 November 2010 in Washington, …

Pursuing Medawar’S Challenge For Full Replacement, Martin Stephens

Experimentation Collection

In 1969, Nobel Prize-winning scientist Peter Medawar predicted that scientific innovation would someday fully replace the use of animals in biomedical research. Medawar correctly forecast the leveling off and subsequent decline in animal use in the last quarter of the 20th century – a period of remarkable innovation in the life sciences. A 2007 report by the U.S. National Academy of Sciences, Toxicity Testing in the 21st Century, proposed a strategy that is likely to replace all routine animal use in toxicology with innovative methods within one to two decades. Replacing animal use throughout biomedical research is more challenging given …

A Roadmap For The Development Of Alternative (Non-Animal) Methods For Systemic Toxicity Testing, David Basketter, Harvey Clewell, Ian Kimber, Annamaria Rossi, Bas Blaauboer, Robert Burrier, Mardas Daneshian, Chantra Eskes, Alan Goldberg, Nina Hasiwa, Sebastian Hoffmann, Joanna Jaworska, Thomas B. Knudsen, Robert Landsiedel, Marcel Leist, Paul Locke, Gavin Maxwell, James Mckim, Emily Mcvey, Gladys Ouédraogo, Grace Patlewicz, Olavi Pelkonen, Erwin Roggen, Costanza Rovida, Irmela Ruhdel, Michael Schwarz, Andreas Schepky, Greet Schoeters, Nigel Skinner, Kerstin Trentz, Marian Turner, Philippe Vanparys, James Yager, Joanne Zurlo, Thomas Hartung

Biomedical Research and Alternative Methods Collection

Systemic toxicity testing forms the cornerstone for the safety evaluation of substances. Pressures to move from traditional animal models to novel technologies arise from various concerns, including: the need to evaluate large numbers of previously untested chemicals and new products (such as nanoparticles or cell therapies), the limited predictivity of traditional tests for human health effects, duration and costs of current approaches, and animal welfare considerations. The latter holds especially true in the context of the scheduled 2013 marketing ban on cosmetic ingredients tested for systemic toxicity. Based on a major analysis of the status of alternative methods (Adler et …