Laboratory and Basic Science Research Commons^™

Efficient Method For Transfer Of Microinjected Eggs To Mouse Ampulla For Generating Transgenic Mice, Guang Wen, Jin Di, Qian Li, Jianling Chen, Ling Jin, Cheng Wang, Sanqing Xu

Publications and Research

Background: The new method described here is highly efficient in transferring microinjected mouse eggs (MEs) through the bursa membrane of a surrogate mother mouse to the ampulla of the oviduct without damaging the blood vessels on the bursa membrane.

Results: This method causes no loss of blood, and it produces newborn pups/founders from approximately 70% of the transferred MEs, because only a small hole is made on the blood vessel–free area of the bursa membrane and ampulla of the surrogate mother mouse. The infundibulum remains intact. The small hole on the bursa membrane/ ampulla may already heal up before the …

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 …

Diagnosis Of Strongyloides Stercoralis: Detection Of Parasite-Derived Dna In Urine, Nilanjan Lodh, Reynaldo Caro, Shterna Sofer, Alan Scott, Alejandro Krolewiecki, Clive J. Shiff

Clinical Lab Sciences Faculty Research and Publications

Detecting infections of Strongyloides stercoralis is arduous and has low sensitivity. Clinically this is a major problem because chronic infections may disseminate in the host and lead to a life threatening condition. Epidemiologically, S. stercoralis is often missed in surveys as it is difficult to identify by standard stool examination procedures. We present, for the first time, evidence that the infection can be detected in filtered urine samples collected and processed in the field and subsequently assayed for the presence of parasite DNA. Urine specimens (∼40 mL) were collected from 125 test and control individuals living in rural and peri-urban …

Update On Laboratory Diagnosis And Epidemiology Of Trichomonas Vaginalis: You Can Teach An “Old” Dog “New” Trichs, Erik Munson, Maureen Napierala, Kimber L. Munson

Clinical Lab Sciences Faculty Research and Publications

Past viewpoints on Trichomonas vaginalis infection have characterized the associated clinical disease as a “nuisance” condition, with affected demographics largely being older African American females residing in urban centers. The advent of commercial molecular assays specific for T. vaginalis has offered a new outlook on trichomoniasis. Within high-prevalence sexually transmitted infection populations, parasite distribution is not localized to specific population centers, and T. vaginalis prevalence is elevated among both younger and older age groups. Adaptation of these molecular assays can additionally facilitate male screening and subsequent epidemiologic characterization. These findings, combined with associations between T. vaginalis infection and human immunodeficiency …

Introduction To Undergraduate Research Evs 295x, Michael Cerbo

Library Impact Statements

No abstract provided.

Profiling Populations Using Neutral Markers, Major Histocompatibility Complex Genes And Volatile Organic Compounds As Modeled In Equus Caballus Linnaeus, Ketaki Deshpande

FIU Electronic Theses and Dissertations

Assessing the genetics of wild animal populations aims to understand selective pressures, and factors whether it be inbreeding or adaptation, that affect the genome. Although numerous techniques are available for assessing population structure, a major obstacle in studying wild populations is obtaining samples from the animals without having to capture them, which can lead to undue distress and injury. Therefore, biologists often use non-invasive sampling methods (i.e., collection of feces, hair) to extract host DNA. In this study, new DNA extraction protocols were developed that improved the quality and quantity of DNA obtained from fecal matter. Fecal samples aged up …

Arthritis Is Developed In Borrelia-Primed And -Infected Mice Deficient Of Interleukin-17, Joseph Kuo, Thomas F. Warner, Erik Munson, Dean T. Nardelli, Ronald F. Schell

Clinical Lab Sciences Faculty Research and Publications

Interleukin-17 (IL-17) has been shown to participate in the development of Lyme arthritis in experimental mice. For example, neutralization of IL-17 with antibodies inhibits induction of arthritis in Borrelia-primed and -infected C57BL/6 wild-type mice. We hypothesized that mice lacking IL-17 would fail to develop Borrelia-induced arthritis. IL-17-deficient and wild-type C57BL/6 mice were primed with heat-inactivated Borrelia and then infected with viable spirochetes 3 weeks later. No swelling or major histopathological changes of the hind paws were detected in IL-17-deficient or wild-type mice that were primed with Borrelia or infected with viable spirochetes. By contrast, IL-17-deficient and wild-type mice …

Live And Dead Coral Cover On Three Reefs And Public Perceptions Of Degradation Around Almirante Bay, Bocas Del Toro, Phoebe Thompson

Independent Study Project (ISP) Collection

Almirante Bay is a lagoonal body of water off the western Caribbean coast of Panama. The islands within the bay are popular tourist destinations, drawing visitors from around the world to explore beaches and coral reefs. However, the reefs in Almirante Bay are continually exposed to both natural and anthropogenic stressors, such as temperature inversions, boat pollution, sedimentation from dredging, and chemical runoff from coastal banana plantations. Because the economy of the area is heavily dependent on tourism, this creates an interesting dynamic in that the main source of income (tourism) could be detrimental to a main attraction of the …

Benthic Algae And Diatom Communities In Seagrass Meadows Under Three Different Human Impact Regimes In Bocas Del Toro, Panamá, Averyl Cheng

Independent Study Project (ISP) Collection

In Bocas del Toro, Panamá, widespread tourism has been the main source of revenue and has become an increasing threat to seagrass meadows and the organisms they support. This study aimed to investigate and describe algae and diatom communities under three different regimes of anthropogenic disturbance: high, medium and low human impact. The biodiversity was analyzed by measuring the algae and diatom assemblages with Shannon-Weiner’s Biodiversity Index, Evenness Index and Sorensen’s coefficient. The data obtained from these three diversity indices were compared to the areas along an anthropogenic disturbance gradient of high, medium and low impact. A total of 12 …

Acute Respiratory Distress Syndrome Caused By Plasmodium Falciparum Infection, Erik Munson, Patricia G. Luedke, William D. Timm

Clinical Lab Sciences Faculty Research and Publications

No abstract provided.

Gc Verification Of The Spacecraft Atmosphere Monitor, Jessica S. Castro, Richard D. Kidd, Jeffrey D. Hein

STAR Program Research Presentations

International Space Station crew members face the unique challenge of maintaining air quality due to the volatile organic compounds (VOCs) that have the potential to accumulate at unsafe levels. The Spacecraft Atmosphere Monitor (SAM) is a miniature gas chromatograph/mass spectrometer (GCMS) designed to measure major constituents (such as N₂, O₂ and CO₂) and trace VOCs within the cabin of the spacecraft. The gas chromatograph is responsible for separating the sample into its components in order to be characterized. The oven of the gas chromatograph must reach a temperature of 150°C in order to heat constituents …

Contribution Of A Putative Up Element Dna Sequence To The Activity Of A Newly Identified Phage Promoter, Courtney Hamilton

Mahurin Honors College Capstone Experience/Thesis Projects

In transcription, a universal step in gene expression, information from a DNA sequence is copied into RNA. A key component in gene expression is the promoter sequence, a region of DNA to which RNA polymerase binds during the initiation of transcription of downstream genes. Most bacterial promoters contain a -10 and a -35 sequence that are bound by the RNA polymerase. Some promoters also contain an Upstream Promoter (UP) element. UP elements have been shown to boost promoter activity. We recently identified a new promoter in a mutant bacteriophage that grows on a bacterial host that prevents antitermination of phage …

Relationship Between Morphogenesis And Secretion In The Filamentous Fungus Aspergillus Nidulans, Lakshmi Preethi Yerra

School of Biological Sciences: Dissertations, Theses, and Student Research

Filamentous fungi have a long history in biotechnology for the production of food ingredients, pharmaceuticals and enzymes. The advancements made in recent years have earned filamentous fungi such as Aspergillus species a dominant place among microbial cell factories. Although the model fungus A. nidulans has been extensively studied, the genetic and regulatory networks that underlie morphogenesis and development have yet to be fully characterized. The Rho GTPases (Cdc42 and RacA) are one of the most important regulators of the morphogenetic processes among diverse eukaryotic organisms. Although the functions of these GTPases are relatively well-characterized, little is known about their downstream …

A Biochemical Approach To Define The Interactome For Calpain2 In Endothelial Cells, Dorothy E. Croall Phd, Jordan R. Hoffman Ms, Calvin Ph Vary Phd, Sharon L. Ashworth Phd, Billy R. Chase, Jay Beaudoin Ii

Molecular & Biomedical Sciences Faculty Scholarship

Current repositories for protein-protein interactions and high throughput screening methods focus on individual gene products and do not consider the significance of calcium induced conformational changes. These limitations suggest the need for alternative strategies to better define the calpain2 interactome. Affinity capture coupled with LC-MS/MS and proteomic analysis of the recovered proteins provides a powerful approach to identify protein-protein interactions for the heterodimeric calpain2. CAPN2 (rat) was modified to be catalytically incompetent (C105A) and fused with a C-terminal 15 residue peptide optimized for biotinylation by the biotin protein ligase, BirA. The resulting CAPN2*, heterodimerized with truncated CAPNS1, was purified from …

Identification, Characterization, And Life Cycle Of Intein-Associated Homing Endonucleases, Joshua J. Skydel

Honors Scholar Theses

Inteins are molecular parasites that have been identified in unicellular organisms from the three domains of life. The intein self-excises following translation of the host gene, and therefore incurs a fitness cost for its carrier. The symbiotic state of the intein to its host is dependent on the presence or absence of a homing endonuclease domain, which facilitates horizontal transfer of the molecule. Identification of this domain provides information on the evolutionary history of the intein, as well as patterns of horizontal gene transfer in microbial communities. I have therefore developed Hidden Markov Models (HMMs) to identify homing endonuclease domains …

Activation Of Target Gene Expression In Neurons By The C. Elegans Rfx Transcription Factor, Daf-19, Katherine P. Mueller

Lawrence University Honors Projects

DAF-19, the only RFX transcription factor found in C. elegans, is required for the formation of neuronal sensory cilia. Four isoforms of the DAF-19 protein have been reported, and the m86 nonsense (null) mutation affecting all four isoforms has been shown to prevent cilia formation. Transcriptome analyses employing microarrays of L1 and adult stage worms were completed using RNA from daf-19(m86) worms and an isogenic wild type strain to identify additional putative DAF-19 target genes. Using transcriptional fusions with GFP, we compared the expression patterns of several potential gene targets using fluorescence confocal microscopy. Expression patterns were characterized in …

Ocean Acidification And Predator-Prey Relations: Correlating Disruption Of Predator Avoidance With Chemosensory Deficits, Alexandra Fw Sidun, William G. Wright

Student Scholar Symposium Abstracts and Posters

One of the most destructive effects of global climate change is the increased carbon sequestering and consequential acidification of our world’s oceans. The impacts of ocean acidification on marine organisms are still relatively unknown, especially effects on behavioral ecology. Avoiding predation has emerged from recent behavioral ecology literature as a critical feature in the life history of a wide array of animal species; experiments on marine fishes suggest acidic water compromises their predator-avoidance abilities. Recent assays in our lab suggest predator-induced behavior is reduced by weakly acidic water. These experiments do not address the potential factor of generalized malaise caused …

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 …

Application Of Alternative Nucleic Acid Extraction Protocols To Progastro Sscs Assay For Detection Of Bacterial Enteric Pathogens, Erik Munson, Maureen Napierala, Kimber L. Munson, Dorothy Bilbo, Michael A. Schulte

Clinical Lab Sciences Faculty Research and Publications

As an alternative to automated extraction, fecal specimens were processed by investigational lysis/heating (i.e., manual) and by chromatography/centrifugation (i.e., column) methods. ProGastro SSC and Shiga toxin-producing Escherichia coli (i.e., STEC) indeterminate rates for 101 specimens were 1.0% to 3.0% for automated, 11.9% for manual, and 24.8% to 37.6% for column methods. Following freeze-thaw of 247 specimens, indeterminate rates were 1.6% to 2.4% for manual and 0.8 to 5.3% for column methods. Mean processing times for manual and column methods were 30.5 and 69.2 min, respectively. Concordance of investigational methods with automated extraction was ≥98.8%.

Rheb1 Expression In Embryonic And Postnatal Mouse, Qi Tian, James L. Smart, Joachim H. Clement, Yingming Wang, Alex Derkatch, Harald Schubert, Michael V. Danilchik, Daniel L. Marks, Lev M. Federov

Faculty Publications - Department of Biological & Molecular Science

Ras homolog enriched in brain (RHEB1) is a member within the superfamily of GTP-binding proteins encoded by the RAS oncogenes. RHEB1 is located at the crossroad of several important pathways including the insulin-signaling pathways and thus plays an important role in different physiological processes. To understand better the physiological relevance of RHEB1 protein, the expres-sion pattern of RHEB1 was analyzed in both embryonic (at E3.5–E16.5) and adult (1-month old) mice. RHEB1 immu-nostaining and X-gal staining were used for wild-type and Rheb1 gene trap mutant mice, respectively. These inde-pendent methods revealed similar RHEB1 expression pat-terns during both embryonic and postnatal developments. …

Non-Essentiality Of Alr And Muri Genes In Mycobacteria, Philion L. Hoff, Denise Zinniel, Raúl G. Barletta

UCARE Research Products

Amino acids are the building blocks of life. If DNA is the blueprint, amino acids are the lumber that proteins are built with. Proteins are built with left-handed, L- forms of amino acids. Bacteria have an essential cell wall component that happens to be an exception: peptidoglycan. Bacteria have enzymes called racemases that convert L- amino acid forms into right-handed, D- forms. Amino acids participate in many reactions with keto acids. Transaminases allow conversion between amino acids by transfer of an amino group.

Previous reports claimed there is no D-ala transaminase activity in mycobacteria and thus alr and murI genes …

How Nebraska’S Eastern Saline Wetland Native Plant Species Grow In Response To Restoration Methods: Application Of Different Salinity Level Groundwater, Ellen Dolph, Keunyea Song, Amy Burgin, Trenton E. Franz

UCARE Research Products

Nebraska’s Eastern Saline Wetlands are unique ecosystems endemic to the Salt and Rock Creek waters in Lancaster and Saunders County.

They provide an ecosystem services as well as habitat for endangered species such as the state endangered saltwort (Salicornia rubra) and federally endangered Salt Creek tiger beetle (Cicindela nevadica lincolniana).

Over 80 % of the saline wetlands are highly degraded and in recent years, the Saline Wetland Conservation Partnership has formed to conserve and restore the remaining saline wetland fragments, but there is limited information about inland saline wetland restoration. Purpose: Investigate techniques to better conserve …

Tom Curran, Phd, Frs, Named Executive Director Of Children's Research Institute, Children's Mercy Hospital

Our Story Continues

Tom Curran, PhD, FRS, has been named Chief Scientific Officer and Executive Director of the Children’s Mercy Children’s Research Institute (CRI), effective Feb. 1 [2016]

The Children’s Research Institute at Children’s Mercy Kansas City was established last year to build on the hospital’s century-long history in pediatric research and to focus the hospital’s future research efforts in four key areas:

• Pediatric Genomic Medicine
• Clinical Pharmacology
• Health Services and Outcomes
• Health Care Delivery

Biographical Feature: Clyde Thornsberry, Ph.D., Erik Munson

Clinical Lab Sciences Faculty Research and Publications

No abstract provided.

Suboptimal Trichomonas Vaginalis Antigen Test Performance In A Low-Prevalence Sexually Transmitted Infection Community, Kimber L. Munson, Maureen Napierala, Erik Munson

Clinical Lab Sciences Faculty Research and Publications

Trichomonas vaginalis is the most common nonviral etiology of sexually transmitted infection (STI) worldwide. The OSOM Trichomonas rapid test (OSOM; Sekisui Diagnostics, San Diego, CA) is a rapid surrogate to microscopic analysis in symptomatic patients, but its performance in low-prevalence STI populations has been assessed on a limited basis in the literature. OSOM has widespread usage, as accreditation data from the College of American Pathologists report that over 300 participant laboratories utilize this assay on an annual basis. We sought to characterize the analytical and clinical performance of OSOM in a low-prevalence STI population on the basis of a commercial …

Models For Hsv Shedding Must Account For Two Levels Of Overdispersion, Amalia Magaret

UW Biostatistics Working Paper Series

We have frequently implemented crossover studies to evaluate new therapeutic interventions for genital herpes simplex virus infection. The outcome measured to assess the efficacy of interventions on herpes disease severity is the viral shedding rate, defined as the frequency of detection of HSV on the genital skin and mucosa. We performed a simulation study to ascertain whether our standard model, which we have used previously, was appropriately considering all the necessary features of the shedding data to provide correct inference. We simulated shedding data under our standard, validated assumptions and assessed the ability of 5 different models to reproduce the …

Systematic Errors In Intro Lab Video Analysis, John Zwart, Kayt E. Frisch, Tim Martin

Faculty Work Comprehensive List

In video analysis lab experiments, students frequently find large discrepancies between results based on self-filmed videos and expected values (e.g. for g determined by a fit to projectile motion data). These differences are frequently far larger than the uncertainty calculated from their fit. Using an inexpensive point-and-shoot camera with a 4x optical zoom to record video, we investigated two possible causes of this error: the effect of placing the reference meter stick at a different object-to-camera distance and the effect of the motion of interest being in a plane not perpendicular to the camera lens. When we observed these phenomena …

Isolation And Characterization Of Microbial Community Associated With Diadumene Lineata, The Orange-Striped Sea Anemone, Tiffany Lin, Andrew Weir

Student and Faculty Research Days

The orange-striped sea anemone, Diadumene lineata, is a marine invertebrate that can be found in the Northern Hemisphere. Distribution of Diadumene lineata ranges from Japan to the Gulf of Argentina. This invasive anemone originated from the Pacific Coast of Asia and serves as a host for a diverse microbial community including a photosynthetic alga. The epiphytic bacteria adhere to the outer surface as well as to the gastrovascular cavity of the host. The microbial community associated with Diadumene lineata is relatively unknown. This study attempted to isolate and identify bacteria commonly associated with this invasive marine invertebrate. Diadumene lineata was …

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 …

Pedagogy For The Pedosphere, Loren B. Byrne, Rachel K. Thiet, V Bala Chaudhary

Arts & Sciences Faculty Publications

No abstract provided.