Life Sciences Commons^™

Transcriptional Dynamics During Rhodococcus Erythropolis Infection With Phage Wc1, Dana Willner, Sudip Paudel, Andrew D. Halleran, Grace E. Solini, Veronica Gray, Margaret Saha

Arts & Sciences Articles

Background

Belonging to the Actinobacteria phylum, members of the Rhodococcus genus thrive in soil, water, and even intracellularly. While most species are non-pathogenic, several cause respiratory disease in animals and, more rarely, in humans. Over 100 phages that infect Rhodococcus species have been isolated but despite their importance for Rhodococcus ecology and biotechnology applications, little is known regarding the molecular genetic interactions between phage and host during infection. To address this need, we report RNA-Seq analysis of a novel Rhodococcus erythopolis phage, WC1, analyzing both the phage and host transcriptome at various stages throughout the infection process.

Results

By five …

Mapping The Tidal Marshes Of Coastal Virginia: A Hierarchical Transfer Learning Approach, Zhonghui Lv, Karinna Nunez, Ethan Brewer, Daniel Runfola

Arts & Sciences Articles

Coastal wetlands, especially tidal marshes, play a crucial role in supporting ecosystems and slowing shoreline erosion. Accurate and cost-effective identification and classification of various marsh types, such as high and low marshes, are important for effective coastal management and conservation endeavors. However, mapping tidal marshes is challenging due to heterogeneous coastal vegetation and dynamic tidal influences. In this study, we employ a deep learning segmentation model to automate the identification and classification of tidal marsh communities in coastal Virginia, USA, using seasonal, publicly available satellite and aerial images. This study leverages the combined capabilities of Sentinel-2 and National Agriculture Imagery …

The Helicobacter Pylori Methylome Is Acid-Responsive Due To Regulation By The Two-Component System Arsrs And The Type I Dna Methyltransferase Hsdm1 (Hp0463), Elise H. Zimmerman, Erin L. Ramsey, Katherine E. Hunter, Sarah M. Villadelgado, Celeste M. Phillips, Ryan T. Shipman, Mark H. Forsyth

Arts & Sciences Articles

In addition to its role in genome protection, DNA methylation can regulate gene expression. In this study, we characterized the impact of acidity, phase variation, and the ArsRS TCS on the expression of the Type I m6A DNA methyltransferase HsdM1 (HP0463) of Helicobacter pylori 26695 and their subsequent effects on the methylome. Transcription of hsdM1 increases at least fourfold in the absence of the sensory histidine kinase ArsS, the major acid-sensing protein of H. pylori. hsdM1 exists in the phase-variable operon hsdR1-hsdM1. Phase-locking hsdR1 (HP0464), the restriction endonuclease gene, has significant impacts on the transcription of hsdM1. To determine the …

Seasonal Trends In Lysogeny In An Appalachian Oak-Hickory Forest Soil, Melaina L. Jacoby, Graham D. Hogg, Madelein R. Assaad, Kurt E. Williamson

Arts & Sciences Articles

Since 1989, investigations into viral ecology have revealed how bacteriophages can influence microbial dynamics within ecosystems at global scales. Most of the information we know about temperate phages, which can integrate themselves into the host genome and remain dormant via a process called lysogeny, has come from research in aquatic ecosystems. Soil environments remain under-studied, and more research is necessary to fully understand the range of impacts phage infections have on the soil bacteria they infect. The aims of this study were to compare the efficacy of different prophage-inducing agents and to elucidate potential temporal trends in lysogeny within a …

Host Defense Peptide Piscidin And Yeast-Derived Glycolipid Exhibit Synergistic Antimicrobial Action Through Concerted Interactions With Membranes, Fei Liu, Alexander I. Greenwood, Yawei Xiong, Et Al., Myriam L. Cotten

Arts & Sciences Articles

Developing new antimicrobials as alternatives to conventional antibiotics has become an urgent race to eradicate drug-resistant bacteria and to save human lives. Conventionally, antimicrobial molecules are studied independently even though they can be cosecreted in vivo. In this research, we investigate two classes of naturally derived antimicrobials: sophorolipid (SL) esters as modified yeast-derived glycolipid biosurfactants that feature high biocompatibility and low production cost; piscidins, which are host defense peptides (HDPs) from fish. While HDPs such as piscidins target the membrane of pathogens, and thus result in low incidence of resistance, SLs are not well understood on a mechanistic level. Here, …

Population Genetics Of Transposable Element Load: A Mechanistic Account Of Observed Overdispersion, Gregory Conradi Smith, Ron D. Smith, Joshua Puzey

Arts & Sciences Articles

In an empirical analysis of transposable element (TE) abundance within natural populations of Mimulus guttatus and Drosophila melanogaster, we found a surprisingly high variance of TE count (e.g., variance-to-mean ratio on the order of 10 to 300). To obtain insight regarding the evolutionary genetic mechanisms that underlie the overdispersed population distributions of TE abundance, we developed a mathematical model of TE population genetics that includes the dynamics of element proliferation and purifying selection on TE load. The modeling approach begins with a master equation for a birth-death process and extends the predictions of the classical theory of TE dynamics in …

Use Of Nest Web Cameras And Citizen Science To Quantify Osprey Prey Delivery Rate And Nest Success, Michael H. Academia, Harmony J. Dalgleish

Arts & Sciences Articles

Ospreys (Pandion haliaetus) are obligate piscivores and their nesting success depends on sufficient amounts of fish delivered to the nests during the breeding season. Nests are considered successful when pairs raise a minimum of one young to fledging or near-fledging age. Through web cameras and online broadcasts of Osprey nests, citizen scientists quantified daily number of fish deliveries, nest survival, and nest success. We received and analyzed curated data (one to seven seasons, 2014–2020) from citizen scientist groups representing 19 Osprey web cameras from four countries in North America and Europe. We compared the average and the coefficient …

Transcriptomes Of Electrophysiologically Recorded Dbx1-Derived Respiratory Neurons Of The Prebötzinger Complex In Neonatal Mice, Prajkta S. Kallurkar, Maria Cristina D. Picardo, Yae K. Sugimura, Margaret Saha, Gregory D. Conradi Smith, Christopher A. Del Negro

Arts & Sciences Articles

Breathing depends on interneurons in the preBötzinger complex (preBötC) derived from Dbx1-expressing precursors. Here we investigate whether rhythm- and pattern-generating functions reside in discrete classes of Dbx1 preBötC neurons. In a slice model of breathing with ~ 5 s cycle period, putatively rhythmogenic Type-1 Dbx1 preBötC neurons activate 100–300 ms prior to Type-2 neurons, putatively specialized for output pattern, and 300–500 ms prior to the inspiratory motor output. We sequenced Type-1 and Type-2 transcriptomes and identified differential expression of 123 genes including ionotropic receptors (Gria3, Gabra1) that may explain their preinspiratory activation profiles and Ca2+ signaling ( …

The Annual Cycle For Whimbrel Populations Using The Western Atlantic Flyway, B. D. Watts, F. Smith, C. Hines, L. Duval, Et Al.

Arts & Sciences Articles

Many long-distance migratory birds use habitats that are scattered across continents and confront hazards throughout the annual cycle that may be population-limiting. Identifying where and when populations spend their time is fundamental to effective management. We tracked 34 adult whimbrels (Numenius phaeopus) from two breeding populations (Mackenzie Delta and Hudson Bay) with satellite transmitters to document the structure of their annual cycles. The two populations differed in their use of migratory pathways and their seasonal schedules. Mackenzie Delta whimbrels made long (22,800 km) loop migrations with different autumn and spring routes. Hudson Bay whimbrels made shorter (17,500 km) …

Intertidal Zonation Of Hemichordates In Soft Sediments, Kharis R. Schrage, Juselyn D. Tupik, Jonathan D. Allen

Arts & Sciences Articles

Intertidal zonation of organisms is well studied on rocky shores but less so in soft sediment communities. On rocky shores, communities are two dimensional, with biotic factors such as competition and predation setting the lower bound of a zone, whereas abiotic factors such as desiccation set the upper bound. In soft sediment communities, these patterns persist, but with a dynamic three-dimensional ecosystem occupied by mobile infaunal organisms, zonation can be more difficult to quantify and detect. Hemichordate worms, however, deposit fecal casts at the surface, which can be easily identified and counted, making them a potential model system for identifying …

Limitations Of 16s Rrna Gene Sequencing To Characterize Lactobacillus Species In The Upper Genital Tract, Jessica L. O'Callaghan, Dana Willner, Melissa Buttini, Flavia Huygens, Elise S. Pelzer

Arts & Sciences Articles

The endometrial cavity is an upper genital tract site previously thought as sterile, however, advances in culture-independent, next-generation sequencing technology have revealed that this low-biomass site harbors a rich microbial community which includes multiple Lactobacillus species. These bacteria are considered to be the most abundant non-pathogenic genital tract commensals. Next-generation sequencing of the female lower genital tract has revealed significant variation amongst microbial community composition with respect to Lactobacillus sp. in samples collected from healthy women and women with urogenital conditions. The aim of this study was to evaluate our ability to characterize members of the genital tract microbial community …

Whimbrel Populations Differ In Trans-Atlantic Pathways And Cyclone Encounters, B. D. Watts, F. M. Smith, C. Hines, L. Duval, Et Al.

Arts & Sciences Articles

Each year hundreds of millions of birds cross the Atlantic Ocean during the peak of tropical cyclone activity. The extent and consequences of migrant-storm interactions remain unknown. We tracked whimbrels from two populations (Mackenzie Delta; Hudson Bay) to examine overlap between migration routes and storm activity and both the frequency and consequence of storm encounters. Here we show that Mackenzie Delta and Hudson Bay whimbrels follow different routes across the ocean and experience dramatically different rates of storm encounters. Mackenzie Delta whimbrels departed North America from Atlantic Canada, made long (x¯" role="presentation" style="box-sizing: inherit; display: inline; line-height: normal; word-spacing: normal; …

The Regulatory Network For Petal Anthocyanin Pigmentation Is Shaped By The Myb5a/Negan Transcription Factor In Mimulus, Xingyu Zheng, Kuenzang Om, Kimmy A. Stanton, (...), Gregory D. Conradi Smith, Joshua R. Puzeyn, Et Al.

Arts & Sciences Articles

Much of the visual diversity of angiosperms is due to the frequent evolution of novel pigmentation patterns in flowers. The gene network responsible for anthocyanin pigmentation, in particular, has become a model for investigating how genetic changes give rise to phenotypic innovation. In the monkeyflower genus Mimulus, an evolutionarily recent gain of petal lobe anthocyanin pigmentation in M. luteus var. variegatus was previously mapped to genomic region pla2. Here, we use sequence and expression analysis, followed by transgenic manipulation of gene expression, to identify MYB5a—orthologous to the NEGAN transcriptional activator from M. lewisii—as the gene responsible …

Comparison Of Nutrient Accrual In Constructed Living Shoreline And Natural Fringing Marshes, Randolph Chambers, A. L. Gorsky, Robert Isdell, Molly Mitchell, Donna Marie Bilkovic

Arts & Sciences Articles

Living shoreline marshes are coastal wetlands constructed as alternatives to “hardened shorelines” (e.g., bulkheads, riprap) to mitigate erosion and to allow for landward migration of intertidal habitat as sea level rises. Living shorelines are designed to mimic natural fringing marshes and over time should be sinks for carbon and other nutrients. We collected soil cores and aboveground plant material from 13 pairs of natural fringing marshes and living shoreline marshes of different ages and degree of isolation from more extensive marsh shorescapes to compare nutrient pools and accrual. Although the nutrient content of plants was similar within and between marsh …

Influence Of Introduced Peregrine Falcons On The Distribution Of Red Knots Within A Spring Staging Site, B. D. Watts, Barry R. Truitt

Arts & Sciences Articles

Predator recovery driven by single-species management approaches may lead to conservation conflicts between recovered predators and prey species of conservation concern. As part of an aggressive recovery plan, the Eastern Peregrine Falcon Recovery Team released (1975–1985) 307 captive-reared peregrine falcons (Falco peregrinus) and successfully established a breeding population within the mid-Atlantic Coastal Plain, a physiographic region with no historic breeding population and a critical spring staging area for migratory shorebirds. We examined the influence of resident falcons on the distribution of foraging red knots during spring migration. We conducted weekly aerial surveys (2006–2009) along the Virginia barrier islands …

Linking Monitoring And Data Analysis To Predictions And Decisions For The Range-Wide Eastern Black Rail Status Assessment, Conor P. Mcgowan, Nicole F. Angeli, Whitney A. Beisler, Et Al., F. M. Smith, B. D. Watts

Arts & Sciences Articles

The US Fish and Wildlife Service (USFWS) has initiated a re-envisioned approach for providing decision makers with the best available science and synthesis of that information, called the Species Status Assessment (SSA), for endangered species decision making. The SSA report is a descriptive document that provides decision makers with an assessment of the current and pre - dicted future status of a species. These analyses support all manner of decisions under the US Endangered Species Act, such as listing, reclassification, and recovery planning. Novel scientific analysis and predictive modeling in SSAs could be an important part of rooting conservation decisions …

Allostery In Oligomeric Receptor Models, Gregory D. Conradi Smith

Arts & Sciences Articles

We show how equilibrium binding curves of receptor homodimers can be expressed as rational polynomial functions of the equilibrium binding curves of the constituent monomers, without approximation and without assuming independence of receptor monomers. Using a distinguished spanning tree construction for reduced graph powers, the method properly accounts for thermodynamic constraints and allosteric interactions between receptor monomers (i.e. conformational coupling). The method is completely general; it begins with an arbitrary undirected graph representing the topology of a monomer state-transition diagram and ends with an algebraic expression for the equilibrium binding curve of a receptor oligomer composed of two or more …

Role Of Synaptic Inhibition In The Coupling Of The Respiratory Rhythms That Underlie Eupnea And Sigh Behaviors, Daniel S. Borrus, Cameron J. Grover, Gregory D. Conradi Smith, Christopher A. Del Negro

Arts & Sciences Articles

The preBötzinger complex (preBötC) gives rise to two types of breathing behavior under normal physiological conditions: eupnea and sighing. Here, we examine the neural mechanisms that couple their underlying rhythms. We measured breathing in awake intact adult mice and recorded inspiratory rhythms from the preBötC in neonatal mouse brainstem slice preparations. We show previously undocumented variability in the temporal relationship between sigh breaths or bursts and their preceding eupneic breaths or inspiratory bursts. Investigating the synaptic mechanisms for this variability in vitro, we further show that pharmacological blockade of chloride-mediated synaptic inhibition strengthens inspiratory-to-sigh temporal coupling. These findings contrast …

Development Of Helical, Fish-Inspired Cross-Step Filter For Collecting Harmful Algae, Adam Schroeder, Lauren Marshall, Brian Trease, Anna Becker, S. Laurie Sanderson

Arts & Sciences Articles

A new filter was developed to collect harmful algae colonies by adapting the cross-step filtration structures and mechanisms discovered recently in filter-feeding fish. Extending beyond previously published models that closely emulated the basic morphology of the fish, the new cross-step filter's major innovations are helical slots, radial symmetry, and rotation as an active anti-clogging mechanism. These innovations enable the transport of concentrated particles to the downstream end of the filter. This advance was made possible by recognizing that biologically imposed constraints such as bilateral symmetry do not apply to human-made filters. The use of helical slots was developed in a …

Localization Of Sumo-Modified Proteins Using Fluorescent Sumo-Trapping Proteins, Rui Yin, Catherine Harvey, Diane C. Shakes, Oliver Kerscher

Arts & Sciences Articles

Here we are presenting a novel method to study the sumoylation of proteins and their sub-cellular localization in mammalian cells and nematode oocytes. This method utilizes a recombinant modified SUMO-trapping protein fragment, kmUTAG, derived from the Ulp1 SUMO protease of the stress-tolerant budding yeast Kluyveromyces marxianus. We have adapted the properties of the kmUTAG for the purpose of studying sumoylation in a variety of model systems without the use of antibodies. For the study of SUMO, KmUTAG has several advantages when compared to antibody-based approaches. This stress-tolerant SUMO-trapping reagent is produced recombinantly, it recognizes native SUMO isoforms from many …

Historical Art, Ecology, And Implication, Alan C. Braddock

Arts & Sciences Articles

"For fifteen years, I have researched, published, lectured, and taught about art and ecology, focusing on contemporary contexts as well as historical work produced long before Ernst Haeckel coined “ecology” (Oecologie) in 1866, and prior to the emergence of modern environmentalism..."

A Likelihood Ratio Test For Changes In Homeolog Expression Bias, Ronald D. Smith, Taliesin J. Kinser, Gregory D. Conradi Smith, Joshua R. Puzeyn

Arts & Sciences Articles

Background

Gene duplications are a major source of raw material for evolution and a likely contributor to the diversity of life on earth. Duplicate genes (i.e., homeologs, in the case of a whole genome duplication) may retain their ancestral function, sub- or neofunctionalize, or be lost entirely. A primary way that duplicate genes evolve new functions is by altering their expression patterns. Comparing the expression patterns of duplicate genes gives clues as to whether any of these evolutionary processes have occurred.

Results

We develop a likelihood ratio test for the analysis of the expression ratios of duplicate genes across two …

Origin And Evolution Of The Octoploid Strawberry Genome, Patrick P. Edger, Thomas J. Poorten, ..., Ron D. Smith, Scott J. Teresi, Joshua R. Puzey, Et Al.

Arts & Sciences Articles

Cultivated strawberry emerged from the hybridization of two wild octoploid species, both descendants from the merger of four diploid progenitor species into a single nucleus more than 1 million years ago. Here we report a near-complete chromosome-scale assembly for cultivated octoploid strawberry (Fragaria × ananassa) and uncovered the origin and evolutionary processes that shaped this complex allopolyploid. We identified the extant relatives of each diploid progenitor species and provide support for the North American origin of octoploid strawberry. We examined the dynamics among the four subgenomes in octoploid strawberry and uncovered the presence of a single dominant subgenome with significantly …

Trpm4 Ion Channels In Pre-Bo¨Tzinger Complex Interneurons Are Essential For Breathing Motor Pattern But Not Rhythm, Marina Cristina D. Picardo, Yae K. Sugimura, Kaitlyn E. Dorst, Prajkta S. Kallurkar, Victoria T. Akins, Xingru Ma, Ryoichi Teruyama, Romaine Guinamard, Kaiwen Kam, Margaret Saha, Christopher A. Del Negro

Arts & Sciences Articles

Inspiratory breathing movements depend on pre-Bötzinger complex (preBötC) interneurons that express calcium (Ca2+)-activated nonselective cationic current (ICAN) to generate robust neural bursts. Hypothesized to be rhythmogenic, reducing ICAN is predicted to slow down or stop breathing; its contributions to motor pattern would be reflected in the magnitude of movements (output). We tested the role(s) of ICAN using reverse genetic techniques to diminish its putative ion channels Trpm4 or Trpc3 in preBötC neurons in vivo. Adult mice transduced with Trpm4-targeted short hairpin RNA (shRNA) progressively decreased the tidal volume of breaths yet surprisingly increased breathing frequency, often followed by gasping and …

Long-Term Monitoring Of A Successful Recovery Program Of Peregrine Falcons In Virginia, B. D. Watts, Mitchell A. Byrd, E. K. Mojica, S. Padgett, S. R. Harding, C. A. Koppie

Arts & Sciences Articles

The Peregrine Falcon (Falco peregrinus anatum) was believed to be extirpated as a breeding species in Virginia by the early 1960s. An aggressive restoration program was initiated in 1978 that involved the release of captive-reared birds totaling 115 on the Coastal Plain (1978–1985) and 127 in the Mountain physiographic region (1985–1993). The first occupied territory was established and the first breeding attempt was documented in 1979 and 1982, respectively. We have monitored the size, distribution, reproductive rate, and substrate use of the resulting breeding population (1979–2016). The population proceeded through an establishment phase (1979–1993) driven by releases with an average …

Sumo-Targeted Ubiquitin Ligases (Stubls) Reduce The Toxicity And Abnormal Transcriptional Activity Associated With A Mutant, Aggregation-Prone Fragment Of Huntingtin, Kentaro Ohkuni, Nagesh Pasupala, Jennifer Peek, Oliver Kerscher

Arts & Sciences Articles

Cell viability and gene expression profiles are altered in cellular models of neurodegenerative disorders such as Huntington’s Disease (HD). Using the yeast model system, we show that the SUMO-targeted ubiquitin ligase (STUbL) Slx5 reduces the toxicity and abnormal transcriptional activity associated with a mutant, aggregation-prone fragment of huntingtin (Htt), the causative agent of HD. We demonstrate that expression of an aggregation-prone Htt construct with 103 glutamine residues (103Q), but not the non-expanded form (25Q), results in severe growth defects in slx5Δ and slx8Δ cells. Since Slx5 is a nuclear protein and because Htt expression affects gene transcription, we …

Dbx1 Pre-Bötzinger Complex Interneurons Comprise The Core Inspiratory Oscillator For Breathing In Unanesthetized Adult Mice, Christopher A. Del Negro, Francis Pham

Arts & Sciences Articles

The brainstem pre-Bötzinger complex (preBötC) generates inspiratory breathing rhythms, but which neurons comprise its rhythmogenic core? Dbx1-derived neurons may play the preeminent role in rhythm generation, an idea well founded at perinatal stages of development but incompletely evaluated in adulthood. We expressed archaerhodopsin or channelrhodopsin in Dbx1 preBötC neurons in intact adult mice to interrogate their function. Prolonged photoinhibition slowed down or stopped breathing, whereas prolonged photostimulation sped up breathing. Brief inspiratory-phase photoinhibition evoked the next breath earlier than expected, whereas brief expiratory-phase photoinhibition delayed the subsequent breath. Conversely, brief inspiratory-phase photostimulation increased inspiratory duration and delayed the subsequent breath, …

Dendritic A-Current In Rhythmically Active Prebo¨Tzinger Complex Neurons In Organotypic Cultures From Newborn Mice, Wiktor S. Phillips, Christopher A. Del Negro, Jens K. Rekling

Arts & Sciences Articles

Brainstem neurons in the preBo¨tC generate the oscillatory activity that underlies breathing. PreBo¨tC neurons express voltagedependent currents that can influence inspiratory activity, among which is a transient potassium current (IA) previously identified in a rhythmogenic excitatory subset of type-1 preBo¨tC neurons. We sought to determine whether IA is expressed in the dendrites of preBo¨tC. We found that dendrites of type-1 preBo¨tC neurons indeed express IA, which may aid in shunting sparse non-summating synaptic inputs, while enabling strong summating excitatory inputs to readily pass and thus influence somatic membrane potential trajectory. The subcellular distribution of IA in rhythmically active neurons of …

Physical Modeling Of Vortical Cross-Step Flow In The American Paddlefish, Polyodon Spathula, Hannah Brooks, Grant Emerson Haines, M. Carly Lin, S. Laurie Sanderson

Arts & Sciences Articles

Vortical cross-step filtration in suspension-feeding fish has been reported recently as a novel mechanism, distinct from other biological and industrial filtration processes. Although crossflow passing over backward-facing steps generates vortices that can suspend, concentrate, and transport particles, the morphological factors affecting this vortical flow have not been identified previously. In our 3D-printed models of the oral cavity for ram suspension-feeding fish, the angle of the backward-facing step with respect to the model’s dorsal midline affected vortex parameters significantly, including rotational, tangential, and axial speed. These vortices were comparable to those quantified downstream of the backward-facing steps that were formed by …

Predator-Induced Plasticity In Egg Capsule Deposition In The Mud Snail, Tritia Obsoleta, Emily A. Harmon, Jonathan D. Allen

Arts & Sciences Articles

Most marine invertebrates develop in the plankton, where microscopic offspring can avoid abundant benthic predators until settlement. However, at least four phyla of marine invertebrates (Annelida, Mollusca, Nemertea, and Platyhelminthes) deposit benthic egg capsules or masses. Often, these animals possess additional means to protect their young, including chemical or morphological defenses or nonrandom selection of deposition sites. Egg capsule deposition is the dominant reproductive strategy among gastropod molluscs, including the mud snail, Tritia obsoleta. In intertidal and shallow subtidal habitats in New England, the mud snail preferentially deposits egg capsules on blades of eelgrass (Zostera marina), a substrate that stands …