Life Sciences Commons^™

Reproducible Hairy Root Transformation And Spot-Inoculation Methods To Study Root Symbioses Of Pea, Scott R. Clemow, Lindsey Clairmont, Lene H. Madsen, Frédérique C. Guinel

Biology Faculty Publications

Pea has lagged behind other model legumes in the molecular study of nodulation and mycorrhizae-formation because of the difficulty to transform its roots and its poor growth on agar plates. Here we describe for pea 1) a transformation technique which permits the complementation of two known non-nodulating pea mutants, 2) a rhizobial inoculation method which allows the study of early cellular events giving rise to nodule primordia, and 3) a targeted fungal inoculation method which allows us to study short segments of mycorrhizal roots assured to be infected. These tools are certain to advance our knowledge of pea root symbioses.

Quantitative Comparison Of Cis-Regulatory Element (Cre) Activities In Transgenic Drosophila Melanogaster, William A. Rogers, Thomas M. Williams

Biology Faculty Publications

Gene expression patterns are specified by cis-regulatory element (CRE) sequences, which are also called enhancers or cis-regulatory modules. A typical CRE possesses an arrangement of binding sites for several transcription factor proteins that confer a regulatory logic specifying when, where, and at what level the regulated gene(s) is expressed. The full set of CREs within an animal genome encodes the organism′s program for development¹, and empirical as well as theoretical studies indicate that mutations in CREs played a prominent role in morphological evolution^2-4. Moreover, human genome wide association studies indicate that genetic variation in CREs …

Protein Trap Lines Of Drosophila To Demonstrate Spatio-Temporal Localization Of Proteins In An Undergraduate Lab, Oorvashi Roy Puli, Amit Singh

Biology Faculty Publications

The objective of this teaching note is to generate a laboratory exercise, which allows students to get a hands-on experience of a cell biology technique. The short duration of the laboratory classes is the biggest challenge with the development of a cell biology lab for an undergraduate curriculum. Therefore, it is necessary to design a laboratory exercise that enables the students to carry out cell biological assays in the desired time. This laboratory exercise focuses on tracking protein expression levels along a spatial (space) and temporal (time) axis in developing Drosophila melanogaster organ primordium. Here we use the protein trap …

Heart And Ventilatory Measures In Crayfish During Copulation, Richard M. Cooper, Heidi Schapker Finucane, Megan Adami, Robin L. Cooper

Biology Faculty Publications

Monitoring heart rate (HR) and ventilatory rate (VR) during defined sensory stimuli and during aggressive and submissive social interactions has provided additional information of a crayfish's physiological state which is not achieved by behavioral observations. In this study, the HR and VR of crayfish were monitored before, during and after the act of copulation in both heterosexual partners. The female crayfish abruptly reduces HR and VR during copulation but the male maintains HR and VR. After separation from copulation the female HR and VR are elevated, potentially paying back the O₂ debt. The tight relationship with HR and VR …

Molecular Systematics Of The Middle American Genus Hypopachus (Anura: Microhylidae), Eli Greenbaum, Eric N. Smith, Rafael O. De Sá

Biology Faculty Publications

We present the first phylogenetic study on the widespread Middle American microhylid frog genus Hypopachus. Partial sequences of mitochondrial (12S and 16S ribosomal RNA) and nuclear (rhodopsin) genes (1275 bp total) were analyzed from 43 samples of Hypopachus, three currently recognized species of Gastrophryne, and seven arthroleptid, brevicipitid and microhylid outgroup taxa. Maximum parsimony (PAUP), maximum likelihood (RAxML) and Bayesian inference (MrBayes) optimality criteria were used for phylogenetic analyses, and BEAST was used to estimate divergence dates of major clades. Population-level analyses were conducted with the programs NETWORK and Arlequin. Results confirm the placement of Hypopachus …

Laboratory Diagnosis Of Clostridium Difficile Infection: Can Molecular Amplification Methods Move Us Out Of Uncertainty?, Fred C. Tenover, Ellen Jo Baron, Lance R. Peterson, David Persing

Biology Faculty Publications

The laboratory diagnosis of Clostridium difficile infection (CDI) continues to be challenging. Recent guidelines from professional societies in the United States note that enzyme immunoassays for toxins A and B do not have adequate sensitivity to be used alone for detecting CDI, yet the optimal method for diagnosing this infection remains unclear. Nucleic acid amplification tests (NAATs) that target chromosomal toxin genes (usually the toxin B gene, tcdB) show high sensitivity and specificity, provide rapid results, and are amenable to both batch and on-demand testing, but these tests were not universally recommended for routine use in the recent guidelines. …

Linking Fruit Traits To Variation In Predispersal Vertebrate Seed Predation, Insect Seed Predation, And Pathogen Attack, Noelle G. Beckman, Helene C. Muller-Landau

Biology Faculty Publications

The importance of vertebrates, invertebrates, and pathogens for plant communities has long been recognized, but their absolute and relative importance in early recruitment of multiple coexisting tropical plant species has not been quantified. Further, little is known about the relationship of fruit traits to seed mortality due to natural enemies in tropical plants. To investigate the influences of vertebrates, invertebrates, and pathogens on reproduction of seven canopy plant species varying in fruit traits, we quantified reductions in fruit development and seed germination due to vertebrates, invertebrates, and fungal pathogens through experimental removal of these enemies using canopy exclosures, insecticide, and …

Minimal Structural Requirements Of Alkyl Γ-Lactones Capable Of Antagonizing The Cocaine-Induced Motility Decrease In Planarians, Debra Baker, Sean Deats, Peter Boor, James Pruitt, Oné R. Pagán

Biology Faculty Publications

We recently reported that the natural cyclic lactone, parthenolide, and related analogs prevent the expression of behavioral effects induced by cocaine in planarians and that parthenolide’s γ-lactone ring is required for this effect. In the present work, we tested a series of alkyl γ-lactones with varying chain length (1–8 carbons) to determine their ability to antagonize the planarian motility decrease induced by 200 μM cocaine. Alkyl lactones with up to a 4-carbon alkyl chain did not affect planarian motility or antagonized the cocaine-induced motility decrease; only the compound γ-nonalactone (a γ-lactone with a 5-carbon chain) was able to prevent the …

Loss-Of-Function Of Constitutive Expresser Of Pathogenesis Related Genes5 Affects Potassium Homeostasis In Arabidopsis Thaliana, Monica Borghi, Ana Rus, David E. Salt

Biology Faculty Publications

Here, we demonstrate that the reduction in leaf K+ observed in a mutant previously identified in an ionomic screen of fast neutron mutagenized Arabidopsis thaliana is caused by a loss-of-function allele of CPR5, which we name cpr5-3. This observation establishes low leaf K+ as a new phenotype for loss-of-function alleles of CPR5. We investigate the factors affecting this low leaf K+ in cpr5 using double mutants defective in salicylic acid (SA) and jasmonic acid (JA) signalling, and by gene expression analysis of various channels and transporters. Reciprocal grafting between cpr5 and Col-0 was used to determine the relative importance of …

Activation Of Jnk Signaling Mediates Amyloid-Ss- Dependent Cell Death, Meghana Tare, Rohan Modi, Jaison Nainaparampil, Oorvashi Roy Puli, Shimpi Bedi, Pedro Fernandez-Funez, Madhuri Kango-Singh, Amit Singh

Biology Faculty Publications

Background: Alzheimer's disease (AD) is an age related progressive neurodegenerative disorder. One of the reasons for Alzheimer's neuropathology is the generation of large aggregates of Aß42 that are toxic in nature and induce oxidative stress, aberrant signaling and many other cellular alterations that trigger neuronal cell death. However, the exact mechanisms leading to cell death are not clearly understood.

Methodology/Principal Findings: We employed a Drosophila eye model of AD to study how Aß42 causes cell death. Misexpression of higher levels of Aß42 in the differentiating photoreceptors of fly retina rapidly induced aberrant cellular phenotypes and cell death. We found that …

No Evidence Of Association Between Hiv-1 And Malaria In Populations With Low Hiv-1 Prevalence, Diego F. Cuadros, Adam J. Branscum, Gisela García-Ramos

Biology Faculty Publications

BACKGROUND: The geographic overlap between HIV-1 and malaria has generated much interest in their potential interactions. A variety of studies have evidenced a complex HIV-malaria interaction within individuals and populations that may have dramatic effects, but the causes and implications of this co-infection at the population level are still unclear. In a previous publication, we showed that the prevalence of malaria caused by the parasite Plasmodium falciparum is associated with HIV infection in eastern sub-Saharan Africa. To complement our knowledge of the HIV-malaria co-infection, the objective of this work was to assess the relationship between malaria and HIV prevalence in …

Effect Of Variable Transmission Rate On The Dynamics Of Hiv In Sub-Saharan Africa, Diego F. Cuadros, Phillip H. Crowley, Ben Augustine, Sarah L. Stewart, Gisela García-Ramos

Biology Faculty Publications

BACKGROUND: The cause of the high HIV prevalence in sub-Saharan Africa is incompletely understood, with heterosexual penile-vaginal transmission proposed as the main mechanism. Heterosexual HIV transmission has been estimated to have a very low probability; but effects of cofactors that vary in space and time may substantially alter this pattern.

METHODS: To test the effect of individual variation in the HIV infectiousness generated by co-infection, we developed and analyzed a mathematical sexual network model that simulates the behavioral components of a population from Malawi, as well as the dynamics of HIV and the co-infection effect caused by other infectious diseases, …

The Cellular Model Of Eastern Oyster (Grassostrea Virginica) Shell Formation, Andrew S. Mount, Neeraj V. Gohad, Karolyn Hansen, Mary Beth Johnstone, Douglas C. Hansen

Biology Faculty Publications

No abstract provided.

The Role Of Crassostrea Virginica Hemocytes In Shell Formation: Ex Vivo Mineral Deposition By Cultured Hemocytes, Emily A. Untener, Douglas C. Hansen, Karolyn Hansen

Biology Faculty Publications

Abstract of Technical Paper Presented at the 103rd Annual Meeting National Shellfisheries Association Baltimore, Maryland March 27–31, 2011

Origin Of Amphibian And Avian Chromosomes By Fission, Fusion, And Retention Of Ancestral Chromosomes, Stephen R. Voss, D. Kevin Kump, Srikrishna Putta, Nathan Pauly, Anna Reynolds, Rema J. Henry, Saritha Basa, John A. Walker, Jeramiah J. Smith

Biology Faculty Publications

Amphibian genomes differ greatly in DNA content and chromosome size, morphology, and number. Investigations of this diversity are needed to identify mechanisms that have shaped the evolution of vertebrate genomes. We used comparative mapping to investigate the organization of genes in the Mexican axolotl (Ambystoma mexicanum), a species that presents relatively few chromosomes (n = 14) and a gigantic genome (>20 pg/N). We show extensive conservation of synteny between Ambystoma, chicken, and human, and a positive correlation between the length of conserved segments and genome size. Ambystoma segments are estimated to be four to 51 times longer than homologous …

Disseminated Neoplasia And Clam Populations In A Canadian National Park-Kouchibouguac National Park., S. Anne Boettger, M J. Abgrall, N Taraska, L Leblanc, E Tremblay

Biology Faculty Publications

No abstract provided.

Disseminated Neoplasia In Mya Arenaria Linked To Chronic Heavy Metal Pollution., S. Anne Boettger, N G. Taraska, J Joseph, C W. Walker

Biology Faculty Publications

No abstract provided.

Survival, Breeding Frequency, And Migratory Orientation In The Jefferson Salamander, Ambystoma Jeffersonianum, Stephen P. De Lisle, Kristine L. Grayson

Biology Faculty Publications

Accurate estimates of demographic parameters, such as survival and breeding frequency, are necessary for the conservation and management of animal populations. Additionally, life-history data are required for gaining an empirical understanding of the ecology of natural populations. We monitored a population of Jefferson Salamanders (Ambystoma jeffersonianum) breeding in a permanent mountain-top pond at the southern limit of this species’ geographic range in Virginia over four years. We used closed multistate mark-recapture models with Pollock's robust design to estimate the demographic parameters of this population. Additionally, we used point-of-capture data to compare the orientation of migrations into and out …

Oak Galls: A Strange Biology Indeed!, W. John Hayden

Biology Faculty Publications

Anyone who takes the time to look closely at several branches of oak will soon find one or another peculiar anomaly among the leaves and twigs. One can easily find structures resembling Ping-Pong balls, hard knots, fluffy tufts, horns—either single or clustered, or irregular thickenings, to mention just a few possibilities. These abnormal growths are galls, structures caused by the presence of small insect larvae living inside the tissue of the plant. Galls can be found on a wide variety of plants. They are common, for example, on the stems of goldenrods, and the leaves of maples, but oaks are …

Mechanical Test Methods For Assessing Porcine Carotid And Uterine Artery Burst Pressure Following Ex Vivo Ultrasonic Ligature Seal And Transection, Carissa M. Krane, Margaret Pinnell, Courtney Gardner, Mercedes Thompson, James Coleman, Robert Wilkens

Biology Faculty Publications

A test method was developed to identify those variables important for assessing the performance of ultrasonic surgical devices in ex vivo ligature sealing of porcine carotid and uterine arteries. Ruggedness testing using a small sample size in pilot experiments was conducted using a newly developed test method in an effort to assess the usefulness of this methodology and to identify test variables that might warrant further testing. The development of this test method included the use of a custom-designed prototypic tension device for load-controlled ex vivo vessel stretching during saline perfusion and subsequent seal and transection of porcine arteries with …

Cooperative And Antagonistic Contributions Of Two Heterochromatin Proteins To Transcriptional Regulation Of The Drosophila Sex Determination Decision, Hui Li, Janel Rodriguez, Youngdong Yoo, Momin Mohammed Shareef, Ramakrishna Badugu, Jamila I. Horabin, Rebecca Kellum

Biology Faculty Publications

Eukaryotic nuclei contain regions of differentially staining chromatin (heterochromatin), which remain condensed throughout the cell cycle and are largely transcriptionally silent. RNAi knockdown of the highly conserved heterochromatin protein HP1 in Drosophila was previously shown to preferentially reduce male viability. Here we report a similar phenotype for the telomeric partner of HP1, HOAP, and roles for both proteins in regulating the Drosophila sex determination pathway. Specifically, these proteins regulate the critical decision in this pathway, firing of the establishment promoter of the masterswitch gene, Sex-lethal (Sxl). Female-specific activation of this promoter, Sxl(Pe), is essential to females, as it provides SXL …

Yeast Bax Inhibitor, Bxi1p, Is An Er-Localized Protein That Links The Unfolded Protein Response And Programmed Cell Death In Saccharomyces Cerevisiae, Nicanor Austriaco, James Cebulski, Joshua Malouin, Nathan Pinches, Vincent Cascio

Biology Faculty Publications

Bax inhibitor-1 (BI-1) is an anti-apoptotic gene whose expression is upregulated in a wide range of human cancers. Studies in both mammalian and plant cells suggest that the BI-1 protein resides in the endoplasmic reticulum and is involved in the unfolded protein response (UPR) that is triggered by ER stress. It is thought to act via a mechanism involving altered calcium dynamics. In this paper, we provide evidence that the Saccharomyces cerevisiae protein encoded by the open reading frame, YNL305C, is a bona fide homolog for BI-1. First, we confirm that yeast cells from two different strain backgrounds lacking YNL305C, …

Rna Interference In Marine And Freshwater Sponges: Actin Knockdown In Tethya Wilhelma And Ephydatia Muelleri By Ingested Dsrna Expressing Bacteria, Ajna S. Rivera, April L. Hill, Et. Al.

Biology Faculty Publications

Background: The marine sponge Tethya wilhelma and the freshwater sponge Ephydatia muelleri are emerging model organisms to study evolution, gene regulation, development, and physiology in non-bilaterian animal systems. Thus far, functional methods (i.e., loss or gain of function) for these organisms have not been available.

Results: We show that soaking developing freshwater sponges in double-stranded RNA and/or feeding marine and freshwater sponges bacteria expressing double-stranded RNA can lead to RNA interference and reduction of targeted transcript levels. These methods, first utilized in C. elegans, have been adapted for the development and feeding style of easily cultured marine and freshwater poriferans. …

Life History Benefits Of Residency In A Partially Migrating Pond-Breeding Amphibian, Kristine L. Grayson, Larissa L. Bailey, Henry M. Wilbur

Biology Faculty Publications

Species with partial migration, where a portion of a population migrates and the other remains residential, provide the opportunity to evaluate conditions for migration and test mechanisms influencing migratory decisions. We conducted a five-year study of two populations of red-spotted newts (Notophthalmus viridescens), composed of individuals that either remain as residents in the breeding pond over the winter or migrate to the terrestrial habitat. We used multistate mark-recapture methods to (1) test for differences in survival probability between migrants and residents, (2) determine if migrants breed every year or skip opportunities for reproduction, and (3) estimate the frequency …

Ecosystem Management Of The Missouri River From Gavins Point Dam To Ponca State Park, Nebraska, Robert S. Nebel

Biology Faculty Publications

•Authorized by a 1978 amendment to the National Parks and Recreation Act (PL 95- 625) which amended the Wild and Scenic Rivers Act of 1968 (PL 90-542)

• Corps is authorized to construct recreational development, bank stabilization, and other recreational river features as necessary to support the values for which the river was designated

• Life-of-project funding ceiling of $21 million; approximately $3.2 million spent to date

• The Corps and National Park Service signed a 1980 Cooperative Agreement outlining each agency's responsibilities

• The General Management Plan has recently been updated (1999) with an environmental emphasis

White Oak Part Of Global Oak Presence, W. John Hayden

Biology Faculty Publications

As we act locally celebrating white oak, Quercus alba, as the 2011 VNPS Wildflower of the Year, it is perhaps appropriate to think globally for a few moments and consider the breadth of diversity encompassed by the oaks. Quercus is a big genus, easily the largest in its family, the Fagaceae. Approximately 400 species of oak are known, and they are widely distributed in the northern hemisphere. We tend to think of oaks as temperate zone trees, but in the New World, their range extends south through the mountains of Central America to Colombia and in the Old World, …

Endo-Porter–Mediated Delivery Of Phosphorodiamidate Morpholino Oligos (Pmos) In Erythrocyte Suspension Cultures From Cope's Gray Treefrog Hyla Chrysoscelis, Venkateshwar Mutyam, Matthew V. Puccetti, James Frisbie, David L. Goldstein, Carissa M. Krane

Biology Faculty Publications

Cope's gray treefrog, Hyla chrysoscelis, is a freeze-tolerant anuran that accumulates cryoprotective glycerol during cold acclimation. H. chrysoscelis erythrocytes express the aquaglyceroporin HC-3, which facilitates transmembrane glycerol and water movement. Aquaglyceroporins have no pharmacological inhibitors, and no genetic knockout tools currently exist for H. chrysoscelis. A phosphorodiamidate morpholino oligo (PMO)–mediated expression knockdown approach was therefore pursued to provide a model for testing the role of HC-3. We describe a novel procedure optimized for specific, efficient knockdown of HC-3 expression in amphibian erythrocyte suspensions cultured at nonmammalian physiological temperatures using Endo-Porter. Our protocol includes three critical components: pre-incubation at 37°C, …

Design Of A Factorial Experiment With Randomization Restrictions To Assess Medical Device Performance On Vascular Tissue, Wiebke Diestelkamp, Carissa M. Krane, Margaret Pinnell

Biology Faculty Publications

Background: Energy-based surgical scalpels are designed to efficiently transect and seal blood vessels using thermal energy to promote protein denaturation and coagulation. Assessment and design improvement of ultrasonic scalpel performance relies on both in vivo and ex vivo testing. The objective of this work was to design and implement a robust, experimental test matrix with randomization restrictions and predictive statistical power, which allowed for identification of those experimental variables that may affect the quality of the seal obtained ex vivo.

Methods: The design of the experiment included three factors: temperature (two levels); the type of solution used to perfuse the …

Use Of Shallow Estuarine Habitats By Nekton In The Mobile-Tensaw River Delta, Alabama, Lawrence P. Rozas, John F. Valentine, Charles Martin, Latina Steele

Biology Faculty Publications

We compared nekton density, composition, and biomass in fall 2009 and spring 2010 among three major habitat types (marsh, SAV=submerged aquatic vegetation dominated by Vallisneria americana, SNB=shallow nonvegetated bottom) commonly found throughout the Mobile-Tensaw River Delta (MTD) using 1-m2drop samplers. In the sampling design, habitat selection was based on vegetation composition. Sample locations (TR=Tensaw River, CB=Chocolatta Bay, and BC=Below Causeway) were selected based on their degree of tidal connectivity with the wider estuary (BC > TR > CB). Nekton distributional patterns varied among both locations and habitat types. Species richness was greater at BC than CB. The young of most …

A Complement Receptor C5a Antagonist Regulates Epithelial To Mesenchymal Transition And Crystallin Expression After Lens Cataract Surgery In Mice, Rinako Suetsugu-Maki, Nobuyasu Maki, Timothy P. Fox, Kenta Nakamura, Richard Cowper.Solari, Craig R. Tomlinson, Hongchang Qu, John D. Lambris, Panagiotis A. Tsonis

Biology Faculty Publications

Purpose: To evaluate the effects of complement employing a mouse model for secondary cataract. Methods: The role of complement receptor C5a (CD88) was evaluated after cataract surgery in mice. An antagonist specific to C5a receptor was administered intraperitoneally to mice. Epithelial to mesenchymal transition (EMT) was evaluated by alpha-smooth muscle actin (α-SMA) staining and proliferation by bromodeoxyuridine (5-bromo-2'- deoxyuridine, BrdU) incorporation. Gene expression patterns was examined by microarray analysis and quantitative polymerase chain reaction (QPCR). Results: We found that administration of a C5aR antagonist in C57BL/6J mice decreases EMT, as evidenced by α-SMA expression, and cell proliferation. Gene expression by …