2024-03-28T11:23:33Zhttps://vtechworks.lib.vt.edu/server/oai/requestoai:vtechworks.lib.vt.edu:10919/858212023-04-14T17:49:45Zcom_10919_23829com_10919_5553col_10919_23830
Becker, Matthew H.
Harris, Reid N.
Biological Sciences
2018-11-12T20:11:11Z
2018-11-12T20:11:11Z
2010-06-04
e10957
http://hdl.handle.net/10919/85821
https://doi.org/10.1371/journal.pone.0010957
5
6
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Chytridiomycosis, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), is an infectious disease that causes population declines of many amphibians. Cutaneous bacteria isolated from redback salamanders, Plethodon cinereus, and mountain yellow-legged frogs, Rana muscosa, inhibit the growth of Bd in vitro. In this study, the bacterial community present on the skin of P. cinereus individuals was investigated to determine if it provides protection to salamanders from the lethal and sub-lethal effects of chytridiomycosis. When the cutaneous bacterial community was reduced prior to Bd exposure, salamanders experienced a significantly greater decrease in body mass, which is a symptom of the disease, when compared to infected individuals with a normal bacterial community. In addition, a greater proportion of infected individuals with a reduced bacterial community experienced limb-lifting, a behavior seen only in infected individuals. Overall, these results demonstrate that the cutaneous bacterial community of P. cinereus provides protection to the salamander from Bd and that alteration of this community can change disease resistance. Therefore, symbiotic microbes associated with this species appear to be an important component of its innate skin defenses.
Published version
application/pdf
en
PLOS
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Cutaneous Bacteria of the Redback Salamander Prevent Morbidity Associated with a Lethal Disease
PLOS ONE
Article - Refereed
Text
ORIGINAL
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journal.pone.0010957.PDF
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2023-04-14 17:49:45.558
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
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VTechWorks
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oai:vtechworks.lib.vt.edu:10919/792302022-02-26T22:17:37Zcom_10919_8195com_10919_25799com_10919_23829com_10919_5553col_10919_78882col_10919_23830
Tornwall, Brett M.
Sokol, Eric R.
Skelton, James
Brown, Bryan L.
Biological Sciences
2017-09-20T18:21:30Z
2017-09-20T18:21:30Z
2015-02-09
2017-09-20T18:21:30Z
Tornwall, B.; Sokol, E.; Skelton, J.; Brown, B.L. Trends in Stream Biodiversity Research since the River Continuum Concept. Diversity 2015, 7, 16-35.
http://hdl.handle.net/10919/79230
https://doi.org/10.3390/d7010016
Lotic environments contain a disproportionate amount of biodiversity given their relatively small proportion of the worldwide landscape. We conducted a systematic literature search of research directed towards understanding factors that influence biodiversity in lotic habitats, published in 31 major ecological and freshwater science journals from 1981 to 2014. Our goal was to characterize emergent themes in research successes and identify important areas in need of study. We show an overwhelming taxonomic bias favoring studies of macroinvertebrates and fish, and a paucity in studies of other important groups such as bacteria and fungi. While most studies assessed habitat variables that affect diversity at a local scale, there has been a recent push to investigate regional drivers of beta and gamma diversity. Several factors were consistently found to be important drivers of diversity including local habitat type, hydrologic variables, disturbance, and stream morphometry. Others such as nutrients and chemical variables showed mixed support. Species interactions, dispersal, and evolutionary processes were rarely considered but show promise as fruitful areas for future study. We suggest that researchers should give increased attention to diversity drivers at different scales as well as take advantage of new molecular techniques to address questions regarding organismal diversity in streams.
Published version
application/pdf
en
MDPI
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
stream ecology
river
microbial diversity
biodiversity
review
metacommunity
Trends in Stream Biodiversity Research since the River Continuum Concept
Diversity
Article - Refereed
Text
ORIGINAL
diversity-07-00016.pdf
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oai:vtechworks.lib.vt.edu:10919/993532022-02-26T22:17:37Zcom_10919_23829com_10919_5553col_10919_23830
Lind, Craig
Moore, Ignacio T.
Akçay, Çağlar
Vernasco, Ben J.
Lorch, Jeffrey M.
Farrell, Terence M.
Biological Sciences
2020-07-14T12:55:15Z
2020-07-14T12:55:15Z
2018-03
1522-2152
http://hdl.handle.net/10919/99353
https://doi.org/10.1086/695747
91
2
29286254
1537-5293
Snake fungal disease (SFD) is an emerging threat to snake populations in the United States. Fungal pathogens are often associated with a physiological stress response mediated by the hypothalamo-pituitary-adrenal axis (HPA), and afflicted individuals may incur steep coping costs. The severity of SFD can vary seasonally; however, little is known regarding (1) how SFD infection relates to HPA activity and (2) how seasonal shifts in environment, life history, or HPA activity may interact to drive seasonal patterns of infection severity and outcomes. To test the hypothesis that SFD is associated with increased HPA activity and to identify potential environmental or physiological drivers of seasonal infection, we monitored baseline corticosterone, SFD infection severity, foraging success, body condition, and reproductive status in a field-active population of pigmy rattlesnakes. Both plasma corticosterone and the severity of clinical signs of SFD peaked in the winter. Corticosterone levels were also elevated in the fall before the seasonal rise in SFD severity. Severely symptomatic snakes were in low body condition and had elevated corticosterone levels compared to moderately infected and uninfected snakes. The monthly mean severity of SFD in the population was negatively related to population-wide estimates of body condition and temperature measured in the precedent month and positively correlated with corticosterone levels measured in the precedent month. Symptomatic females were less likely to enter reproductive bouts compared to asymptomatic females. We propose the hypothesis that the seasonal interplay among environment, host energetics, and HPA activity initiates trade-offs in the fall that drive the increase in SFD prevalence, symptom severity, and decline in condition observed in the population through winter.
Public domain – authored by a U.S. government employee
application/pdf
en
Creative Commons CC0 1.0 Universal Public Domain Dedication
http://creativecommons.org/publicdomain/zero/1.0/
glucocorticoid
hypothalamo-pituitary-adrenal axis
allostatic load
conservation physiology
Ophidiomyces ophiodiicola
cort-fitness hypothesis
Patterns of Circulating Corticosterone in a Population of Rattlesnakes Afflicted with Snake Fungal Disease: Stress Hormones as a Potential Mediator of Seasonal Cycles in Disease Severity and Outcomes
Physiological and Biochemical Zoology
Article - Refereed
Text
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open.access
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oai:vtechworks.lib.vt.edu:10919/736312024-03-13T14:09:53Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553col_10919_70873col_10919_23830
Ciliberto, Andrea
Capuani, Fabrizio
Tyson, John J.
Biological Sciences
2016-12-09T21:34:32Z
2016-12-09T21:34:32Z
2007-03-01
1553-734X
http://hdl.handle.net/10919/73631
https://doi.org/10.1371/journal.pcbi.0030045
3
3
In metabolic networks, metabolites are usually present in great excess over the enzymes that catalyze their interconversion, and describing the rates of these reactions by using the Michaelis–Menten rate law is perfectly valid. This rate law assumes that the concentration of enzyme–substrate complex (C) is much less than the free substrate concentration (S0). However, in protein interaction networks, the enzymes and substrates are all proteins in comparable concentrations, and neglecting C with respect to S0 is not valid. Borghans, DeBoer, and Segel developed an alternative description of enzyme kinetics that is valid when C is comparable to S0. We extend this description, which Borghans et al. call the total quasi-steady state approximation, to networks of coupled enzymatic reactions. First, we analyze an isolated Goldbeter–Koshland switch when enzymes and substrates are present in comparable concentrations. Then, on the basis of a real example of the molecular network governing cell cycle progression, we couple two and three Goldbeter–Koshland switches together to study the effects of feedback in networks of protein kinases and phosphatases. Our analysis shows that the total quasi-steady state approximation provides an excellent kinetic formalism for protein interaction networks, because (1) it unveils the modular structure of the enzymatic reactions, (2) it suggests a simple algorithm to formulate correct kinetic equations, and (3) contrary to classical Michaelis–Menten kinetics, it succeeds in faithfully reproducing the dynamics of the network both qualitatively and quantitatively.
Published version
463 - 472 (10) page(s)
English
PLOS
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000246191000012&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Biochemical Research Methods
Mathematical & Computational Biology
Biochemistry & Molecular Biology
XENOPUS-OOCYTE EXTRACTS
M-PHASE CONTROL
CELL-CYCLE
KINETICS
BISTABILITY
HYSTERESIS
CASCADES
Modeling networks of coupled enzymatic reactions using the total quasi-steady state approximation
PLOS Computational Biology
Article - Refereed
/Virginia Tech
/Virginia Tech/All T&R Faculty
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Science/COS T&R Faculty
/Virginia Tech/University Distinguished Professors
ORIGINAL
Modeling networks of coupled enzymatic reactions using the total quasi-steady state approximation.pdf
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oai:vtechworks.lib.vt.edu:10919/950652021-10-08T16:43:28Zcom_10919_8195com_10919_25799com_10919_23829com_10919_5553col_10919_78882col_10919_23830
Doubek, Jonathan P.
Campbell, Kylie L.
Lofton, Mary E.
McClure, Ryan P.
Carey, Cayelan C.
Biological Sciences
2019-10-25T16:40:46Z
2019-10-25T16:40:46Z
2019-10-19
2019-10-25T14:07:52Z
Doubek, J.P.; Campbell, K.L.; Lofton, M.E.; McClure, R.P.; Carey, C.C. Hypolimnetic Hypoxia Increases the Biomass Variability and Compositional Variability of Crustacean Zooplankton Communities. Water 2019, 11, 2179.
http://hdl.handle.net/10919/95065
https://doi.org/10.3390/w11102179
In freshwater lakes and reservoirs, climate change and eutrophication are increasing the occurrence of low-dissolved oxygen concentrations (hypoxia), which has the potential to alter the variability of zooplankton seasonal dynamics. We sampled zooplankton and physical, chemical and biological variables (e.g., temperature, dissolved oxygen, and chlorophyll <i>a</i>) in four reservoirs during the summer stratified period for three consecutive years. The hypolimnion (bottom waters) of two reservoirs remained oxic throughout the entire stratified period, whereas the hypolimnion of the other two reservoirs became hypoxic during the stratified period. Biomass variability (measured as the coefficient of the variation of zooplankton biomass) and compositional variability (measured as the community composition of zooplankton) of crustacean zooplankton communities were similar throughout the summer in the oxic reservoirs; however, biomass variability and compositional variability significantly increased after the onset of hypoxia in the two seasonally-hypoxic reservoirs. The increase in biomass variability in the seasonally-hypoxic reservoirs was driven largely by an increase in the variability of copepod biomass, while the increase in compositional variability was driven by increased variability in the dominance (proportion of total crustacean zooplankton biomass) of copepod taxa. Our results suggest that hypoxia may increase the seasonal variability of crustacean zooplankton communities.
Published version
application/pdf
en
MDPI
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
aggregate variability
anoxia
calanoid copepod
cladoceran
climate change
cyclopoid copepod
global change
lake
plankton
reservoir
Hypolimnetic Hypoxia Increases the Biomass Variability and Compositional Variability of Crustacean Zooplankton Communities
Water
Article - Refereed
Text
ORIGINAL
water-11-02179.pdf
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oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/95065
2021-10-08 16:43:28.265
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/986122020-10-22T03:35:27Zcom_10919_23829com_10919_5553col_10919_23830
Hsu, Bryan B.
Way, Jeffrey C.
Silver, Pamela A.
Biological Sciences
2020-05-29T14:24:59Z
2020-05-29T14:24:59Z
2020-01-28
2379-5077
e00013-20
http://hdl.handle.net/10919/98612
https://doi.org/10.1128/mSystems.00013-20
5
1
31992629
Elimination or alteration of select members of the gut microbiota is key to therapeutic efficacy. However, the complexity of these microbial inhabitants makes it challenging to precisely target bacteria. Here, we deliver exogenous genes to specific bacteria by genomic integration of temperate phage for long-lasting modification. As a real-world therapeutic test, we engineered lambda phage to transcriptionally repress Shiga toxin by using genetic hybrids between lambda and other lambdoid phages to overcome resistance encoded by the virulence-expressing prophage. We show that a single dose of engineered phage propagates throughout the bacterial community and reduces Shiga toxin production in an enteric mouse model of infection without markedly affecting bacterial concentrations. Our work reveals a new framework for transferring functions to bacteria within their native environment. IMPORTANCE With the increasing frequency of antibiotic resistance, it is critical to explore new therapeutic strategies for treating bacterial infections. Here, we use a temperate phage, i.e., one that integrates itself into the bacterial genome, to neutralize the expression of a virulence factor by modifying bacterial function at the genetic level. We show that Shiga toxin production can be significantly reduced in vitro and in the mammalian gut. Alternative to traditional applications of phage therapy that rely on killing bacteria, our genetics-based antivirulence approach introduces a new framework for treating bacterial infections.
Bill & Melinda Gates Foundation through the Grand Challenges Explorations Initiative [OPP1150555]; Defense Advanced Research Program Agency (DARPA BRICS)United States Department of DefenseDefense Advanced Research Projects Agency (DARPA) [HR0011-15-C-0094]; Harvard Medical School; Rosenbloom postdoctoral fellowship
This project was funded by the Bill & Melinda Gates Foundation through the Grand Challenges Explorations Initiative (OPP1150555), the Defense Advanced Research Program Agency (DARPA BRICS HR0011-15-C-0094), and funds from Harvard Medical School. B.B.H. received support from the Rosenbloom postdoctoral fellowship.
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Shiga toxin
bacteriophage
microbiome
antivirulence
Stable Neutralization of a Virulence Factor in Bacteria Using Temperate Phage in the Mammalian Gut
mSystems
Article - Refereed
Text
StillImage
ORIGINAL
mSystems-2020-Hsu-e00013-20.full.pdf
mSystems-2020-Hsu-e00013-20.full.pdf
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10919/98612
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/98612
2020-10-22 03:35:27.182
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1050762022-02-26T16:44:49Zcom_10919_8195com_10919_25799com_10919_23829com_10919_5553com_10919_91913com_10919_23198col_10919_78882col_10919_23830col_10919_91914
King, D. Ryan
Sedovy, Meghan W.
Leng, Xinyan
Xue, Jianxiang
Lamouille, Samy Y.
Koval, Michael
Isakson, Brant E.
Johnstone, Scott R.
Fralin Biomedical Research Institute
Biological Sciences
2021-09-27T12:27:23Z
2021-09-27T12:27:23Z
2021-09-22
2021-09-25T23:33:17Z
King, D.R.; Sedovy, M.W.; Leng, X.; Xue, J.; Lamouille, S.; Koval, M.; Isakson, B.E.; Johnstone, S.R. Mechanisms of Connexin Regulating Peptides. Int. J. Mol. Sci. 2021, 22, 10186.
http://hdl.handle.net/10919/105076
https://doi.org/10.3390/ijms221910186
Gap junctions (GJ) and connexins play integral roles in cellular physiology and have been found to be involved in multiple pathophysiological states from cancer to cardiovascular disease. Studies over the last 60 years have demonstrated the utility of altering GJ signaling pathways in experimental models, which has led to them being attractive targets for therapeutic intervention. A number of different mechanisms have been proposed to regulate GJ signaling, including channel blocking, enhancing channel open state, and disrupting protein-protein interactions. The primary mechanism for this has been through the design of numerous peptides as therapeutics, that are either currently in early development or are in various stages of clinical trials. Despite over 25 years of research into connexin targeting peptides, the overall mechanisms of action are still poorly understood. In this overview, we discuss published connexin targeting peptides, their reported mechanisms of action, and the potential for these molecules in the treatment of disease.
Published version
application/pdf
en
MDPI
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Mechanisms of Connexin Regulating Peptides
International Journal of Molecular Science
Article - Refereed
Text
ORIGINAL
ijms-22-10186.pdf
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10919/105076
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/105076
2022-02-26 16:44:49.136
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/468152022-05-03T13:49:21Zcom_10919_23829com_10919_5553col_10919_23830
Cheever, B. M.
Webster, Jackson R.
Bilger, E. E.
Thomas, S. A.
Biological Sciences
2014-03-27T13:05:59Z
2014-03-27T13:05:59Z
2013-07
2014-03-11
B. M. Cheever, J. R. Webster, E. E. Bilger, and S. A. Thomas 2013. The relative importance of exogenous and substrate-derived nitrogen for microbial growth during leaf decomposition. Ecology 94:1614-1625. http://dx.doi.org/10.1890/12-1339.1
0012-9658
http://hdl.handle.net/10919/46815
http://www.esajournals.org/doi/pdf/10.1890/12-1339.1
https://doi.org/10.1890/12-1339.1
Heterotrophic microbes colonizing detritus obtain nitrogen (N) for growth by assimilating N from their substrate or immobilizing exogenous inorganic N. Microbial use of these two pools has different implications for N cycling and organic matter decomposition in the face of the global increase in biologically available N. We used sugar maple leaves labeled with N-15 to differentiate between microbial N that had been assimilated from the leaf substrate (enriched with N-15) or immobilized from the water (natural abundance N-15:N-14) in five Appalachian streams ranging in ambient NO3-N concentrations from about 5 to 900 g NO3-N/L. Ambient NO3- concentration increased sugar maple decomposition rate but did not influence the proportion of microbial N derived from substrate or exogenous pools. Instead, these proportions were strongly influenced by the percentage of detrital ash-free dry mass (AFDM) remaining. Substrate-derived N made up a large proportion of the microbial N after the first 24 h in all streams. Detrital and microbial isotopic N-15 signatures approached that of the water as decomposition progressed in all streams, suggesting that exogenous N may be the predominant source of N for meeting microbial requirements even when exogenous N concentrations are low. Our results support predictions of more rapid decomposition of organic matter in response to increased N availability and highlight the tight coupling of processes driving microbial N cycling and organic matter decomposition.
NSF DEB-0823293, DEB-02218001, DEB-478406
en_US
Ecological Society of America
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Ecological Society of America
n-15
assimilation
chloroform fumigation
heterotrophic microbes
immobilization
nitrogen availability
nitrogen cycling
organic matter
decomposition
streams
forest litter decomposition
soil-nitrogen
woodland stream
chloroform
fumigation
phosphorus dynamics
foliar litter
nutrient
carbon
release
lignin
The relative importance of exogenous and substrate-derived nitrogen for microbial growth during leaf decomposition
Ecology
Article - Refereed
ORIGINAL
12-1339%2E1.pdf
12-1339%2E1.pdf
Main article
application/pdf
571068
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/e8416bb7-1f5c-4e88-a0b3-4c48e6a270d2/download
214ac546dfdff8770a3a8e8cf442a384
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TEXT
12-1339%2E1.pdf.txt
12-1339%2E1.pdf.txt
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text/plain
50737
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MD5
4
10919/46815
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/46815
2022-05-03 13:49:21.712
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1050462022-06-16T17:38:38Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553col_10919_70873col_10919_23830
Aylward, Frank O.
Santoro, Alyson E.
Biological Sciences
2021-09-23T12:46:27Z
2021-09-23T12:46:27Z
2020-05-01
2021-09-23T12:46:23Z
ARTN e00415-20 (Article number)
2379-5077
5/3/e00415-20 (PII)
http://hdl.handle.net/10919/105046
https://doi.org/10.1128/mSystems.00415-20
5
3
Aylward, Frank [0000-0002-1279-4050]
32546674
2379-5077
The Thaumarchaeota is a diverse archaeal phylum comprising numerous lineages that play key roles in global biogeochemical cycling, particularly in the ocean. To date, all genomically characterized marine thaumarchaea are reported to be chemolithoautotrophic ammonia oxidizers. In this study, we report a group of putatively heterotrophic marine thaumarchaea (HMT) with small genome sizes that is globally abundant in the mesopelagic, apparently lacking the ability to oxidize ammonia. We assembled five HMT genomes from metagenomic data and show that they form a deeply branching sister lineage to the ammonia-oxidizing archaea (AOA). We identify this group in metagenomes from mesopelagic waters in all major ocean basins, with abundances reaching up to 6% of that of AOA. Surprisingly, we predict the HMT have small genomes of ∼1 Mbp, and our ancestral state reconstruction indicates this lineage has undergone substantial genome reduction compared to other related archaea. The genomic repertoire of HMT indicates a versatile metabolism for aerobic chemoorganoheterotrophy that includes a divergent form III-a RuBisCO, a 2M respiratory complex I that has been hypothesized to increase energetic efficiency, and a three-subunit heme-copper oxidase complex IV that is absent from AOA. We also identify 21 pyrroloquinoline quinone (PQQ)-dependent dehydrogenases that are predicted to supply reducing equivalents to the electron transport chain and are among the most highly expressed HMT genes, suggesting these enzymes play an important role in the physiology of this group. Our results suggest that heterotrophic members of the Thaumarchaeota are widespread in the ocean and potentially play key roles in global chemical transformations. Importance: It has been known for many years that marine Thaumarchaeota are abundant constituents of dark ocean microbial communities, where their ability to couple ammonia oxidation and carbon fixation plays a critical role in nutrient dynamics. In this study, we describe an abundant group of putatively heterotrophic marine Thaumarchaeota (HMT) in the ocean with physiology distinct from those of their ammonia-oxidizing relatives. HMT lack the ability to oxidize ammonia and fix carbon via the 3-hydroxypropionate/4-hydroxybutyrate pathway but instead encode a form III-a RuBisCO and diverse PQQ-dependent dehydrogenases that are likely used to conserve energy in the dark ocean. Our work expands the scope of known diversity of Thaumarchaeota in the ocean and provides important insight into a widespread marine lineage.
Published version
20 page(s)
application/pdf
en
American Society for Microbiology
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000576704600022&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Life Sciences & Biomedicine
Microbiology
Thaumarchaeota
marine archaea
TACK
PQQ-dehydrogenase
RuBisCO
QUINOPROTEIN GLUCOSE-DEHYDROGENASE
ALCOHOL-DEHYDROGENASE
CRYSTAL-STRUCTURE
ESCHERICHIA-COLI
ARCHAEA
EVOLUTION
DIVERSITY
INSIGHTS
DATABASE
TREE
Heterotrophic Thaumarchaea with Small Genomes Are Widespread in the Dark Ocean
mSystems
Article - Refereed
Article
Journal
Text
/Virginia Tech
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/All T&R Faculty
/Virginia Tech/Science/COS T&R Faculty
SWORD
sword-2021-09-23T08:46:23.original.xml
Original SWORD entry document
application/octet-stream
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MD5
1
ORIGINAL
Heterotrophic Thaumarchaea with Small Genomes Are Widespread in the Dark Ocean.pdf
Heterotrophic Thaumarchaea with Small Genomes Are Widespread in the Dark Ocean.pdf
Published version
application/pdf
3715696
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MD5
2
TEXT
Heterotrophic Thaumarchaea with Small Genomes Are Widespread in the Dark Ocean.pdf.txt
Heterotrophic Thaumarchaea with Small Genomes Are Widespread in the Dark Ocean.pdf.txt
Extracted text
text/plain
94875
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MD5
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THUMBNAIL
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Heterotrophic Thaumarchaea with Small Genomes Are Widespread in the Dark Ocean.pdf.jpg
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MD5
4
10919/105046
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/105046
2022-06-16 17:38:38.768
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/932932020-10-22T03:35:27Zcom_10919_23829com_10919_5553col_10919_23830
Zhang, Yao
Geng, Shuo
Prasad, G.L.
Li, Liwu
Biological Sciences
2019-08-29T12:25:42Z
2019-08-29T12:25:42Z
2018-10-04
Zhang Y, Geng S, Prasad GL and Li L (2018) Suppression of Neutrophil Antimicrobial Functions by Total Particulate Matter From Cigarette Smoke. Front. Immunol. 9:2274. doi: 10.3389/fimmu.2018.02274
http://hdl.handle.net/10919/93293
https://doi.org/10.3389/fimmu.2018.02274
9
Chronic cigarette smoking is widely known to alter immune functions and compromise host defense against microbial infection. Neutrophils play an essential role in the immune defense against microbial pathogens and also participate in the development of the inflammatory responses. However, there is limited information about the effects of cigarette smoking on neutrophil response. In this study, cultured bonemarrow neutrophils were exposed to total particulate matter (TPM) from cigarette smoke. We found that TPM not only reduced LPS-induced TNFa production, but also suppressed neutrophil bactericidal activity. We also observed that TPM priming reduced the expression of NADPH oxidase component gp91 and iNOS, molecules important for bacterial killing. Mechanistically, we documented that TPM-primed neutrophils have reduced STAT1 activation following subsequent LPS challenge. STAT1 is a key transcription factor responsible for the expression of inflammatory genes as well as gp91 and iNOS. Collectively, reduced STAT1 activation and reduced NADPH oxidase/iNOS may potentially explain the compromised anti-microbial function of TPM-programmed neutrophils. Taken together, our findings reveal that the key innate immune neutrophil is subject to reprogramming by smoking to adopt an immune-suppressed state, potentially responsible for chronic smoking-mediated immunosuppression.
R. J. Reynolds Tobacco Company
9 pages
application/pdf
en
Frontiers
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
neutrophil programming
TPM
immune-modulation
NADPH oxidase
inflammation
Suppression of Neutrophil Antimicrobial Functions by Total Particulate Matter From Cigarette Smoke
Frontiers in Immunology
Article - Refereed
Text
ORIGINAL
fimmu-09-02274.pdf
fimmu-09-02274.pdf
application/pdf
2975661
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MD5
1
TEXT
fimmu-09-02274.pdf.txt
fimmu-09-02274.pdf.txt
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35516
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2
THUMBNAIL
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fimmu-09-02274.pdf.jpg
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MD5
4
10919/93293
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/93293
2020-10-22 03:35:27.221
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/739592023-11-29T11:23:54Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553com_10919_24230com_10919_5532com_10919_25796col_10919_70873col_10919_23830col_10919_24305col_10919_25797
Naumenko, Anastasia N.
Timoshevskiy, Vladimir A.
Kinney, Nicholas A.
Kokhanenko, Alina A.
deBruyn, Becky S.
Lovin, Diane D.
Stegniy, Vladimir N.
Severson, David W.
Sharakhov, Igor V.
Sharakhova, Maria V.
Biological Sciences
Entomology
Fralin Life Sciences Institute
2017-01-05T22:13:15Z
2017-01-05T22:13:15Z
2015-03-13
1932-6203
http://hdl.handle.net/10919/73959
https://doi.org/10.1371/journal.pone.0115737
10
3
The genome assembly of southern house mosquito Cx. quinquefasciatus is represented by a high number of supercontigs with no order or orientation on the chromosomes. Although cytogenetic maps for the polytene chromosomes of this mosquito have been developed, their utilization for the genome mapping remains difficult because of the low number of high quality spreads in chromosome preparations. Therefore, a simple and robust mitotic-chromosome-based approach for the genome mapping of Cx. quinquefasciatus still needs to be developed. In this study, we performed physical mapping of 37 genomic supercontigs using fluorescent in situ hybridization on mitotic chromosomes from imaginal discs of 4th instar larvae. The genetic linkage map nomenclature was adopted for the chromosome numbering based on the direct positioning of 58 markers that were previously genetically mapped. The smallest, largest, and intermediate chromosomes were numbered as 1, 2, and 3, respectively. For idiogram development, we analyzed and described in detail the morphology and proportions of the mitotic chromosomes. Chromosomes were subdivided into 19 divisions and 72 bands of four different intensities. These idiograms were used for mapping the genomic supercontigs/genetic markers. We also determined the presence of length polymorphism in the q arm of sex-determining chromosome 1 in Cx. quinquefasciatus related to the size of ribosomal locus. Our physical mapping and previous genetic linkage mapping resulted in the chromosomal assignment of 13% of the total genome assembly to the chromosome bands.We provided the first detailed description, nomenclature, and idiograms for the mitotic chromosomes of Cx. quinquefasciatus. Further application of the approach developed in this study will help to improve the quality of the southern house mosquito genome.
Published version
? - ? (13) page(s)
application/pdf
en
PLOS
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000351277500002&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
quantitative trait loci
mosquito aedes-aegypti
malaria parasite susceptibility
tubule polytene chromosomes
anopheles-gambiae
pipiens complex
linkage map
diptera
culicidae
sequence
Mitotic-Chromosome-Based Physical Mapping of the Culex quinquefasciatus Genome
PLOS ONE
Article - Refereed
Text
/Virginia Tech
/Virginia Tech/Agriculture & Life Sciences
/Virginia Tech/Agriculture & Life Sciences/CALS T&R Faculty
/Virginia Tech/Agriculture & Life Sciences/Entomology
/Virginia Tech/All T&R Faculty
/Virginia Tech/Faculty of Health Sciences
ORIGINAL
Mitotic-chromosome-based physical mapping of the Culex quinquefasciatus genome.pdf
Publisher's Version
application/pdf
2437461
https://vtechworks.lib.vt.edu/bitstreams/28311561-be21-4f75-9407-da2fbbd9130a/download
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MD5
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LICENSE
VTUL_Distribution_License_2016_05_09.pdf
application/pdf
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MD5
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TEXT
Mitotic-chromosome-based physical mapping of the Culex quinquefasciatus genome.pdf.txt
Mitotic-chromosome-based physical mapping of the Culex quinquefasciatus genome.pdf.txt
Extracted text
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45023
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IM Thumbnail
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MD5
9
10919/73959
oai:vtechworks.lib.vt.edu:10919/73959
2023-11-29 11:23:54.683
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
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oai:vtechworks.lib.vt.edu:10919/1030632021-04-21T07:16:48Zcom_10919_23829com_10919_5553col_10919_23830
Abdelfattah, Ahmed
Freilich, Shiri
Bartuv, Rotem
Zhimo, V. Yeka
Kumar, Ajay
Biasi, Antonio
Salim, Shoshana
Feygenberg, Oleg
Burchard, Erik
Dardick, Christopher
Liu, Jia
Khan, Awais
Ellouze, Walid
Ali, Shawkat
Spadaro, Davide
Torres, Rosario
Teixido, Neus
Ozkaya, Okan
Buehlmann, Andreas
Vero, Silvana
Mondino, Pedro
Berg, Gabriele
Wisniewski, Michael
Droby, Samir
Biological Sciences
2021-04-20T19:39:27Z
2021-04-20T19:39:27Z
2021-03-18
1462-2912
http://hdl.handle.net/10919/103063
https://doi.org/10.1111/1462-2920.15469
33734550
1462-2920
We present the first worldwide study on the apple (Malus x domestica) fruit microbiome that examines questions regarding the composition and the assembly of microbial communities on and in apple fruit. Results revealed that the composition and structure of the fungal and bacterial communities associated with apple fruit vary and are highly dependent on geographical location. The study also confirmed that the spatial variation in the fungal and bacterial composition of different fruit tissues exists at a global level. Fungal diversity varied significantly in fruit harvested in different geographical locations and suggests a potential link between location and the type and rate of postharvest diseases that develop in each country. The global core microbiome of apple fruit was represented by several beneficial microbial taxa and accounted for a large fraction of the fruit microbial community. The study provides foundational information about the apple fruit microbiome that can be utilized for the development of novel approaches for the management of fruit quality and safety, as well as for reducing losses due to the establishment and proliferation of postharvest pathogens. It also lays the groundwork for studying the complex microbial interactions that occur on apple fruit surfaces.
BARD, Israel- US Binational Agricultural Research and Development Fund [IS-5040-17]; European UnionEuropean Commission [844114]
This research was funded by BARD, Israel- US Binational Agricultural Research and Development Fund, (IS-5040-17) awarded to S.D. and M.W. European Union's Horizon2020 under 'Nurturing excellence by means of cross-border and cross-sector mobility' program for MSCA-IF-2018-Individual Fellowships, grant agreement 844114 [A.A.].
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Global analysis of the apple fruit microbiome: are all apples the same?
Environmental Microbiology
Article - Refereed
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oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/103063
2021-04-21 07:16:48.261
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1032952021-10-08T16:43:28Zcom_10919_8195com_10919_25799com_10919_23829com_10919_5553col_10919_78882col_10919_23830
Kraikivski, Pavel
Academy of Integrated Science
2021-05-14T13:15:47Z
2021-05-14T13:15:47Z
2021-05-08
2021-05-13T14:35:55Z
Kraikivski, P. Implications of Noise on Neural Correlates of Consciousness: A Computational Analysis of Stochastic Systems of Mutually Connected Processes. Entropy 2021, 23, 583.
http://hdl.handle.net/10919/103295
https://doi.org/10.3390/e23050583
Random fluctuations in neuronal processes may contribute to variability in perception and increase the information capacity of neuronal networks. Various sources of random processes have been characterized in the nervous system on different levels. However, in the context of neural correlates of consciousness, the robustness of mechanisms of conscious perception against inherent noise in neural dynamical systems is poorly understood. In this paper, a stochastic model is developed to study the implications of noise on dynamical systems that mimic neural correlates of consciousness. We computed power spectral densities and spectral entropy values for dynamical systems that contain a number of mutually connected processes. Interestingly, we found that spectral entropy decreases linearly as the number of processes within the system doubles. Further, power spectral density frequencies shift to higher values as system size increases, revealing an increasing impact of negative feedback loops and regulations on the dynamics of larger systems. Overall, our stochastic modeling and analysis results reveal that large dynamical systems of mutually connected and negatively regulated processes are more robust against inherent noise than small systems.
Published version
application/pdf
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Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
neural correlates of consciousness
spectral entropy
power spectrum
stochastic modeling
noise in neuronal networks
Implications of Noise on Neural Correlates of Consciousness: A Computational Analysis of Stochastic Systems of Mutually Connected Processes
Entropy
Article - Refereed
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oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/103295
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Creative Commons Attribution 4.0 International
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vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1016192020-12-24T08:30:54Zcom_10919_23829com_10919_5553com_10919_23261com_10919_5539com_10919_79468com_10919_78628col_10919_23830col_10919_23262col_10919_79481
Menon, Nidhi
Dang, Ha X.
Datla, Udaya Sree
Moarefian, Maryam
Lawrence, Christopher B.
Maher, Christopher A.
Jones, Caroline N.
Biological Sciences
Mechanical Engineering
2020-12-23T18:08:33Z
2020-12-23T18:08:33Z
2020-05-21
2047-4830
http://hdl.handle.net/10919/101619
https://doi.org/10.1039/c9bm01481k
8
10
32091043
2047-4849
The tumor microenvironment plays a critical role in the proliferation and chemoresistance of cancer cells. Growth factors (GFs) are known to interact with the extracellular matrix (ECM) via heparin binding sites, and these associations influence cell behavior. In the present study, we demonstrate the ability to define signals presented by the scaffold by pre-mixing growth factors, such as epidermal growth factor, into the heparin-based (HP-B) hydrogel prior to gelation. In the 3D biomimetic microenvironment, breast cancer cells formed spheroids within 24 hours of initial seeding. Despite higher number of proliferating cells in 2D cultures, 3D spheroids exhibited a higher degree of chemoresistance after 72 hours. Further, our RNA sequencing results highlighted the phenotypic changes influenced by solid-phase GF presentation. Wnt/beta-catenin and TGF-beta signaling were upregulated in the cells grown in the hydrogel, while apoptosis, IL2-STAT5 and PI3K-AKT-mTOR signaling were downregulated. With emerging technologies for precision medicine in cancer, this nature of fine-tuning the microenvironment is paramount for cultivation and downstream characterization of primary cancer cells and rare circulating tumor cells (CTCs), and effective screening of chemotherapeutic agents.
National Institute of General Medical Sciences of the National Institutes of HealthUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of General Medical Sciences (NIGMS) [R35GM133610]
We would like to thank Carla Finkielstein for her thoughtful discussions on experimental design and RNA seq data analysis. Research reported in this publication was supported by The National Institute of General Medical Sciences of the National Institutes of Health under award number R35GM133610.
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Heparin-based hydrogel scaffolding alters the transcriptomic profile and increases the chemoresistance of MDA-MB-231 triple-negative breast cancer cells
Biomaterials Science
Article - Refereed
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oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/101619
2020-12-24 08:30:54.269
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Creative Commons Attribution 4.0 International
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https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1151552023-05-24T07:12:06Zcom_10919_23829com_10919_5553col_10919_23830
Webster, Jackson R.
Jackson, C. Rhett
Knoepp, Jennifer D.
Bolstad, Paul V.
2023-05-23T14:36:43Z
2023-05-23T14:36:43Z
2023-01
0885-6087
http://hdl.handle.net/10919/115155
https://doi.org/10.1002/hyp.14812
37
1
1099-1085
Most studies of land use effects on solute concentrations in streams have focused on smaller streams with watersheds dominated by a single land-use type. Using land cover as a proxy for land use, the objective of this study was to determine whether the hydrologically-driven response of solutes to land use in small streams could be scaled up to predict concentrations in larger receiving streams and rivers in the rural area of the Little Tennessee River basin. We measured concentrations of typically limiting nutrients (nitrogen, phosphorus), abundant anions (chloride, sulfate), and base cations in 17 small streams and four larger river sites. In the small streams, total solute concentration was strongly related to land cover -- highest in streams with developed watersheds, lowest in streams with forested watersheds, and streams with agricultural watersheds were in between. In general, the best predictor of solute concentrations in the small streams was forest land cover. We then predicted solute concentrations for the river sites based on the solute--land cover relationships of the small streams using multiple linear regressions. Results were mixed -- some of the predicted river concentrations were close to measured values, others were greater or less than measured concentrations. In general, river concentrations did not scale with land cover-solute relationships found in small tributaries. Measured values of nitrogen solutes in the river sites were greater than predicted, perhaps due to the presence of waste water treatment plants. We attributed other differences between measured and predicted river concentrations to the heterogeneous geochemistry of this mountainous region. The combined complexity of hydrology, geochemistry, and human land-use of this mountainous region make it difficult to scale up from small streams to larger river basins.
National Science Foundation [DEB0823293, DEB9632854, DEB0218001]
Published version
National Science Foundation, Grant/Award Numbers: DEB0823293, DEB9632854,DEB0218001
Public domain – authored by a U.S. government employee
application/pdf
en
Wiley
Public Domain (U.S.)
http://creativecommons.org/publicdomain/mark/1.0/
geochemistry
land use
rural
solutes
stream
watershed
Can small stream solute-land cover relationships predict river solute concentrations?
Hydrological Processes
Article - Refereed
Text
ORIGINAL
WebsterCan.pdf
WebsterCan.pdf
Published version
application/pdf
1902296
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/ea48e7b1-bbeb-4f28-b2bf-28acf1c7d377/download
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MD5
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TEXT
WebsterCan.pdf.txt
WebsterCan.pdf.txt
Extracted text
text/plain
77213
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/3d615de9-11bf-4cb5-8460-5e4c5779831d/download
783064dce5079f93718872ce85c5b39a
MD5
2
THUMBNAIL
WebsterCan.pdf.jpg
WebsterCan.pdf.jpg
IM Thumbnail
image/jpeg
6123
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/8173c9c2-77f6-4d88-a088-05463eeb6d8c/download
e85d7aa62d6f966a1469faff617a74f8
MD5
3
10919/115155
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/115155
2023-05-24 07:12:06.703
http://creativecommons.org/publicdomain/mark/1.0/
Public Domain (U.S.)
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/955172023-11-29T12:06:44Zcom_10919_23829com_10919_5553com_10919_23965com_10919_5557col_10919_23830col_10919_24280
Beck, Michelle L.
Hopkins, William A.
Hallagan, John J.
Jackson, Brian P.
Hawley, Dana M.
Biological Sciences
Fish and Wildlife Conservation
2019-11-13T13:49:23Z
2019-11-13T13:49:23Z
2014
Beck ML, Hopkins WA, Hallagan JJ, Jackson BP, Hawley DM (2014) Exposure to residual concentrations of elements from a remediated coal fly ash spill does not adversely influence stress and immune responses of nestling tree swallows. Conserv Physiol 2: doi:10.1093/conphys/cou018.
http://hdl.handle.net/10919/95517
https://doi.org/10.1093/conphys/cou018
2
Anthropogenic activities often produce pollutants that can affect the physiology, growth and reproductive success of wildlife. Many metals and trace elements play important roles in physiological processes, and exposure to even moderately elevated concentrations of essential and non-essential elements could have subtle effects on physiology, particularly during development. We examined the effects of exposure to a number of elements from a coal fly ash spill that occurred in December 2008 and has since been remediated on the stress and immune responses of nestling tree swallows. We found that nestlings at the site of the spill had significantly greater blood concentrations of Cu, Hg, Se and Zn in 2011, but greater concentrations only of Se in 2012, in comparison to reference colonies. The concentrations of elements were below levels of significant toxicological concern in both years. In 2011, we found no relationship between exposure to elements associated with the spill and basal or stress-induced corticosterone concentrations in nestlings. In 2012, we found that Se exposure was not associated with cellmediated immunity based on the response to phytohaemagglutinin injection. However, the bactericidal capacity of nestling plasma had a positive but weak association with blood Se concentrations, and this association was stronger at the spill site. Our results indicate that exposure to these low concentrations of elements had few effects on nestling endocrine and immune physiology. The long-term health consequences of low-level exposure to elements and of exposure to greater element concentrations in avian species require additional study.
This research was funded by a grant from the Tennessee Valley Authority [TVA# 555245] to W.A.H. and D.M.H.
13 pages
application/pdf
en
Oxford University Press
Creative Commons Attribution 3.0 Unported
http://creativecommons.org/licenses/by/3.0/
bactericidal capacity
cell-mediated immunity
element
stress response
tree swallow
Exposure to residual concentrations of elements from a remediated coal fly ash spill does not adversely influence stress and immune responses of nestling tree swallows
Conservation Physiology
Article - Refereed
Text
ORIGINAL
cou018.pdf
cou018.pdf
application/pdf
1137471
https://vtechworks.lib.vt.edu/bitstreams/a2c4af2a-59d7-4610-be76-4401f5317abd/download
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MD5
1
TEXT
cou018.pdf.txt
cou018.pdf.txt
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text/plain
71708
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MD5
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THUMBNAIL
cou018.pdf.jpg
cou018.pdf.jpg
IM Thumbnail
image/jpeg
61096
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MD5
6
10919/95517
oai:vtechworks.lib.vt.edu:10919/95517
2023-11-29 12:06:44.841
http://creativecommons.org/licenses/by/3.0/
Creative Commons Attribution 3.0 Unported
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1155552023-06-29T07:12:06Zcom_10919_24226com_10919_5532com_10919_23829com_10919_5553col_10919_24301col_10919_23830
Salar, Safoura
Ball, Nicolas E.
Baaziz, Hiba
Nix, Jay C.
Sobe, Richard C.
Compton, K. Karl
Zhulin, Igor B.
Brown, Anne M.
Scharf, Birgit E.
Schubot, Florian D.
2023-06-28T12:55:31Z
2023-06-28T12:55:31Z
2023-05
0887-3585
http://hdl.handle.net/10919/115555
https://doi.org/10.1002/prot.26510
37213073
1097-0134
Chemotaxis is a fundamental process whereby bacteria seek out nutrient sources and avoid harmful chemicals. For the symbiotic soil bacterium Sinorhizobium meliloti, the chemotaxis system also plays an essential role in the interaction with its legume host. The chemotactic signaling cascade is initiated through interactions of an attractant or repellent compound with chemoreceptors or methyl-accepting chemotaxis proteins (MCPs). S. meliloti possesses eight chemoreceptors to mediate chemotaxis. Six of these receptors are transmembrane proteins with periplasmic ligand-binding domains (LBDs). The specific functions of McpW and McpZ are still unknown. Here, we report the crystal structure of the periplasmic domain of McpZ (McpZPD) at 2.7 angstrom resolution. McpZPD assumes a novel fold consisting of three concatenated four-helix bundle modules. Through phylogenetic analyses, we discovered that this helical tri-modular domain fold arose within the Rhizobiaceae family and is still evolving rapidly. The structure, offering a rare view of a ligand-free dimeric MCP-LBD, reveals a novel dimerization interface. Molecular dynamics calculations suggest ligand binding will induce conformational changes that result in large horizontal helix movements within the membrane-proximal domains of the McpZPD dimer that are accompanied by a 5 angstrom vertical shift of the terminal helix toward the inner cell membrane. These results suggest a mechanism of transmembrane signaling for this family of MCPs that entails both piston-type and scissoring movements. The predicted movements terminate in a conformation that closely mirrors those observed in related ligand-bound MCP-LBDs.
National Science Foundation; NIH Office of the Director
Published version
National Science Foundation; NIH Office of the Director
application/pdf
en
Wiley
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
chemotaxis
helical tri-modular sensor domain
ligand-binding domain
methyl-accepting chemotaxis protein
piston
scissoring
transmembrane signaling
The structural analysis of the periplasmic domain of Sinorhizobium meliloti chemoreceptor McpZ reveals a novel fold and suggests a complex mechanism of transmembrane signaling
Proteins-Structure Function and Bioinformatics
Article - Refereed
Text
ORIGINAL
SalarStructural.pdf
SalarStructural.pdf
Published version
application/pdf
3297986
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/a3cd264e-158f-4cd4-9d78-d174eb4efc6b/download
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MD5
1
TEXT
SalarStructural.pdf.txt
SalarStructural.pdf.txt
Extracted text
text/plain
64704
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/45a4a7a7-ecf9-413f-ac76-c399545b7276/download
f42eb2e2e0a439f94eb08d3d223b532e
MD5
2
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SalarStructural.pdf.jpg
SalarStructural.pdf.jpg
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MD5
3
10919/115555
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/115555
2023-06-29 07:12:06.694
http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1148222023-08-30T19:29:26Zcom_10919_8195com_10919_25799com_10919_23829com_10919_5553com_10919_24216com_10919_5539com_10919_91913com_10919_23198com_10919_91912col_10919_78882col_10919_23830col_10919_24286col_10919_91914col_10919_91916
Payne, Laura Beth
Abdelazim, Hanaa
Hoque, Maruf
Barnes, Audra
Mironovova, Zuzana
Willi, Caroline E.
Darden, Jordan
Houk, Clifton
Sedovy, Meghan W.
Johnstone, Scott R.
Chappell, John C.
2023-04-27T14:52:59Z
2023-04-27T14:52:59Z
2023-04-21
2023-04-27T13:50:44Z
Payne, L.B.; Abdelazim, H.; Hoque, M.; Barnes, A.; Mironovova, Z.; Willi, C.E.; Darden, J.; Houk, C.; Sedovy, M.W.; Johnstone, S.R.; Chappell, J.C. A Soluble Platelet-Derived Growth Factor Receptor-β Originates via Pre-mRNA Splicing in the Healthy Brain and Is Upregulated during Hypoxia and Aging. Biomolecules 2023, 13, 711.
http://hdl.handle.net/10919/114822
https://doi.org/10.3390/biom13040711
The platelet-derived growth factor-BB (PDGF-BB) pathway provides critical regulation of cerebrovascular pericytes, orchestrating their investment and retention within the brain microcirculation. Dysregulated PDGF Receptor-beta (PDGFRβ) signaling can lead to pericyte defects that compromise blood-brain barrier (BBB) integrity and cerebral perfusion, impairing neuronal activity and viability, which fuels cognitive and memory deficits. Receptor tyrosine kinases such as PDGF-BB and vascular endothelial growth factor-A (VEGF-A) are often modulated by soluble isoforms of cognate receptors that establish signaling activity within a physiological range. Soluble PDGFRβ (sPDGFRβ) isoforms have been reported to form by enzymatic cleavage from cerebrovascular mural cells, and pericytes in particular, largely under pathological conditions. However, pre-mRNA alternative splicing has not been widely explored as a possible mechanism for generating sPDGFRβ variants, and specifically during tissue homeostasis. Here, we found sPDGFRβ protein in the murine brain and other tissues under normal, physiological conditions. Utilizing brain samples for follow-on analysis, we identified mRNA sequences corresponding to sPDGFRβ isoforms, which facilitated construction of predicted protein structures and related amino acid sequences. Human cell lines yielded comparable sequences and protein model predictions. Retention of ligand binding capacity was confirmed for sPDGFRβ by co-immunoprecipitation. Visualizing fluorescently labeled sPDGFRβ transcripts revealed a spatial distribution corresponding to murine brain pericytes alongside cerebrovascular endothelium. Soluble PDGFRβ protein was detected throughout the brain parenchyma in distinct regions, such as along the lateral ventricles, with signals also found more broadly adjacent to cerebral microvessels consistent with pericyte labeling. To better understand how sPDGFRβ variants might be regulated, we found elevated transcript and protein levels in the murine brain with age, and acute hypoxia increased sPDGFRβ variant transcripts in a cell-based model of intact vessels. Our findings indicate that soluble isoforms of PDGFRβ likely arise from pre-mRNA alternative splicing, in addition to enzymatic cleavage mechanisms, and these variants exist under normal physiological conditions. Follow-on studies will be needed to establish potential roles for sPDGFRβ in regulating PDGF-BB signaling to maintain pericyte quiescence, BBB integrity, and cerebral perfusion—critical processes underlying neuronal health and function, and in turn, memory and cognition.
Published version
application/pdf
en
MDPI
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
A Soluble Platelet-Derived Growth Factor Receptor-β Originates via Pre-mRNA Splicing in the Healthy Brain and Is Upregulated during Hypoxia and Aging
Biomolecules
Article - Refereed
Text
ORIGINAL
biomolecules-13-00711.pdf
Published version
application/pdf
4259070
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/f619e035-f0a8-4184-8616-57b6126a8f08/download
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MD5
1
LICENSE
license.txt
license.txt
text/plain; charset=utf-8
0
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MD5
2
SWORD
packager-644a7db2.zip
SWORD deposit package
application/octet-stream
4090407
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/b887284c-ad8b-4ba1-925e-8f218501f8f4/download
53e6032983e7aa4181e726805d8d5cb4
MD5
3
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biomolecules-13-00711.pdf.txt
biomolecules-13-00711.pdf.txt
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text/plain
94095
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/58bc8bea-1d08-4531-a702-1b99ebb19013/download
e46e6fe835bfc89359c6a47d6edf21bb
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biomolecules-13-00711.pdf.jpg
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https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/9c1b6c3e-1943-4ee5-b5c1-ae397322d799/download
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MD5
5
10919/114822
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/114822
2023-08-30 19:29:26.98
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1023722023-11-29T12:16:56Zcom_10919_23829com_10919_5553com_10919_23967com_10919_5557col_10919_23830col_10919_24279
Thomas, R. Quinn
Figueiredo, Renato J.
Daneshmand, Vahid
Bookout, Bethany J.
Puckett, Laura K.
Carey, Cayelan C.
Forest Resources and Environmental Conservation
Biological Sciences
2021-02-15T20:03:29Z
2021-02-15T20:03:29Z
2020-11
0043-1397
e2019WR026138
http://hdl.handle.net/10919/102372
https://doi.org/10.1029/2019WR026138
56
11
1944-7973
Freshwater ecosystems are experiencing greater variability due to human activities, necessitating new tools to anticipate future water quality. In response, we developed and deployed a real-time iterative water temperature forecasting system (FLARE-Forecasting Lake And Reservoir Ecosystems). FLARE is composed of water temperature and meteorology sensors that wirelessly stream data, a data assimilation algorithm that uses sensor observations to update predictions from a hydrodynamic model and calibrate model parameters, and an ensemble-based forecasting algorithm to generate forecasts that include uncertainty. Importantly, FLARE quantifies the contribution of different sources of uncertainty (driver data, initial conditions, model process, and parameters) to each daily forecast of water temperature at multiple depths. We applied FLARE to Falling Creek Reservoir (Vinton, Virginia, USA), a drinking water supply, during a 475-day period encompassing stratified and mixed thermal conditions. Aggregated across this period, root mean square error (RMSE) of daily forecasted water temperatures was 1.13 degrees C at the reservoir's near-surface (1.0 m) for 7-day ahead forecasts and 1.62 degrees C for 16-day ahead forecasts. The RMSE of forecasted water temperatures at the near-sediments (8.0 m) was 0.87 degrees C for 7-day forecasts and 1.20 degrees C for 16-day forecasts. FLARE successfully predicted the onset of fall turnover 4-14 days in advance in two sequential years. Uncertainty partitioning identified meteorology driver data as the dominant source of uncertainty in forecasts for most depths and thermal conditions, except for the near-sediments in summer, when model process uncertainty dominated. Overall, FLARE provides an open-source system for lake and reservoir water quality forecasting to improve real-time management.
U.S. National Science FoundationNational Science Foundation (NSF) [CNS-1737424, DEB-1753639, DBI-1933016, DEB-1926050, DEB-1926388]; Virginia Tech Global Change Center; Fralin Life Sciences Institute
This work was supported by the U.S. National Science Foundation (CNS-1737424, DEB-1753639, DBI-1933016, DEB-1926050, and DEB-1926388); the Virginia Tech Global Change Center; and Fralin Life Sciences Institute. We thank our Ecological Forecasting seminar students, the Smart Reservoir team, and Ecological Forecasting Initiative (EFI) colleagues for helpful feedback on the project; the Western Virginia Water Authority for their long-term support and access to field sites; and Mary Lofton, Ryan McClure, and Whitney Woelmer for their critical help in the field.
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
data assimilation
ecological forecasting
ensemble Kalman filter
FLARE
General Lake Model
water temperature
A Near-Term Iterative Forecasting System Successfully Predicts Reservoir Hydrodynamics and Partitions Uncertainty in Real Time
Water Resources Research
Article - Refereed
Text
StillImage
ORIGINAL
2019WR026138.pdf
2019WR026138.pdf
application/pdf
2304817
https://vtechworks.lib.vt.edu/bitstreams/ee1687d3-dcf4-4bef-bd41-9b6f4d3a9b2c/download
2415e03b264875f2c684f78ee0a000bf
MD5
1
TEXT
2019WR026138.pdf.txt
2019WR026138.pdf.txt
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text/plain
86986
https://vtechworks.lib.vt.edu/bitstreams/8fdb023f-54bb-45b1-bc53-0937ab0edd67/download
35e9a8ebbfa67e28e9dfcbb404d7acbe
MD5
2
THUMBNAIL
2019WR026138.pdf.jpg
2019WR026138.pdf.jpg
IM Thumbnail
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67597
https://vtechworks.lib.vt.edu/bitstreams/877c9deb-e7ed-4e16-9c50-855a2864482d/download
8c1121659760e6c2cc92b0f117cc343a
MD5
4
10919/102372
oai:vtechworks.lib.vt.edu:10919/102372
2023-11-29 12:16:56.612
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1052252022-06-17T18:58:49Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553col_10919_70873col_10919_23830
Martinez-Gutierrez, Carolina Alejandra
Aylward, Frank O.
2021-10-11T12:42:17Z
2021-10-11T12:42:17Z
2019-10-01
2021-10-11T12:42:15Z
1759-6653
5572122 (PII)
http://hdl.handle.net/10919/105225
https://doi.org/10.1093/gbe/evz201
11
10
Aylward, Frank [0000-0002-1279-4050]
31539038
1759-6653
Marine microorganisms inhabiting nutrient-depleted waters play critical roles in global biogeochemical cycles due to their abundance and broad distribution. Many of these microbes share similar genomic features including small genome size, low % G + C content, short intergenic regions, and low nitrogen content in encoded amino acid residue side chains (N-ARSC), but the evolutionary drivers of these characteristics are unclear. Here, we compared the strength of purifying selection across the Marinimicrobia, a candidate phylum which encompasses a broad range of phylogenetic groups with disparate genomic features, by estimating the ratio of nonsynonymous and synonymous substitutions (dN/dS) in conserved marker genes. Our analysis reveals that epipelagic Marinimicrobia that exhibit features consistent with genome streamlining have significantly lower dN/dS values when compared with their mesopelagic counterparts. We also found a significant positive correlation between median dN/dS values and % G + C content, N-ARSC, and intergenic region length. We did not identify a significant correlation between dN/dS ratios and estimated genome size, suggesting the strength of selection is not a primary factor shaping genome size in this group. Our findings are generally consistent with genome streamlining theory, which postulates that many genomic features of abundant epipelagic bacteria are the result of adaptation to oligotrophic nutrient conditions. Our results are also in agreement with previous findings that genome streamlining is common in epipelagic waters, suggesting that microbes inhabiting this region of the ocean have been shaped by strong selection together with prevalent nutritional constraints characteristic of this environment.
Published version
Pages 2887-2894
8 page(s)
application/pdf
en
Oxford University Press
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000504448300016&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
Creative Commons Attribution-NonCommercial 4.0 International
http://creativecommons.org/licenses/by-nc/4.0/
Life Sciences & Biomedicine
Evolutionary Biology
Genetics & Heredity
genome streamlining
purifying selection
evolutionary genomics
dN/dS ratio
Marinimicrobia
uncultured picoplankton
CLOSELY-RELATED BACTERIAL
POPULATION-SIZE
EVOLUTION
ALIGNMENTS
DIVERGENCE
DYNAMICS
PATTERNS
TOOL
Developmental Biology
0601 Biochemistry and Cell Biology
0603 Evolutionary Biology
0604 Genetics
Bacteria
Genetic Markers
Evolution, Molecular
Genome, Bacterial
Selection, Genetic
Strong Purifying Selection Is Associated with Genome Streamlining in Epipelagic Marinimicrobia
Genome Biology and Evolution
Article - Refereed
Article
Journal
Text
2019-09-08
/Virginia Tech
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/All T&R Faculty
/Virginia Tech/Science/COS T&R Faculty
SWORD
sword-2021-10-11T08:42:15.original.xml
Original SWORD entry document
application/octet-stream
130
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MD5
1
ORIGINAL
Strong Purifying Selection Is Associated with Genome Streamlining in Epipelagic Marinimicrobia.pdf
Strong Purifying Selection Is Associated with Genome Streamlining in Epipelagic Marinimicrobia.pdf
Published version
application/pdf
480271
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/b13bfeaf-6c7d-4509-8631-d2225e27c850/download
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MD5
2
TEXT
Strong Purifying Selection Is Associated with Genome Streamlining in Epipelagic Marinimicrobia.pdf.txt
Strong Purifying Selection Is Associated with Genome Streamlining in Epipelagic Marinimicrobia.pdf.txt
Extracted text
text/plain
35628
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/08948f0d-7462-426f-af99-a3232f4ec54b/download
2382936a7dd8319a979775b1d21bd86f
MD5
3
THUMBNAIL
Strong Purifying Selection Is Associated with Genome Streamlining in Epipelagic Marinimicrobia.pdf.jpg
Strong Purifying Selection Is Associated with Genome Streamlining in Epipelagic Marinimicrobia.pdf.jpg
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5923
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eed60fcb0687c8519935c1acfcf30154
MD5
4
10919/105225
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/105225
2022-06-17 18:58:49.829
http://creativecommons.org/licenses/by-nc/4.0/
Creative Commons Attribution-NonCommercial 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/933922022-02-26T15:58:35Zcom_10919_78629com_10919_78628com_10919_23829com_10919_5553col_10919_78630col_10919_23830
McCullough, Ian M.
Dugan, Hilary A.
Farrell, Kaitlin J.
Morales-Williams, Ana M.
Ouyang, Zutao
Roberts, Derek C.
Scordo, Facundo
Bartlett, Sarah L.
Burke, Samantha M.
Doubek, Jonathan P.
Krivak-Tetley, Flora E.
Skaff, Nicholas K.
Summers, Jamie C.
Weathers, Kathleen C.
Hanson, Paul C.
Biological Sciences
2019-09-05T14:45:56Z
2019-09-05T14:45:56Z
2018-10-24
0304-3800
http://hdl.handle.net/10919/93392
https://doi.org/10.1016/j.ecolmodel.2018.08.009
386
1872-7026
Lakes are active processors of organic carbon (OC) and play important roles in landscape and global carbon cycling. Allochthonous OC loads from the landscape, along with autochthonous OC loads from primary production, are mineralized in lakes, buried in lake sediments, and exported via surface or groundwater outflows. Although these processes provide a basis for a conceptual understanding of lake OC budgets, few studies have integrated these fluxes under a dynamic modeling framework to examine their interactions and relative magnitudes. We developed a simple, dynamic mass balance model for OC, and applied the model to a set of five lakes. We examined the relative magnitudes of OC fluxes and found that long-term (> 10 year) lake OC dynamics were predominantly driven by allochthonous loads in four of the five lakes, underscoring the importance of terrestrially-derived OC in northern lake ecosystems. Our model highlighted seasonal patterns in lake OC budgets, with increasing water temperatures and lake productivity throughout the growing season corresponding to a transition from burial- to respiration-dominated OC fates. Ratios of respiration to burial, however, were also mediated by the source (autochthonous vs. allochthonous) of total OC loads. Autochthonous OC is more readily respired and may therefore proportionally reduce burial under a warming climate, but allochthonous OC may increase burial due to changes in precipitation. The ratios of autochthonous to allochthonous inputs and respiration to burial demonstrate the importance of dynamic models for examining both the seasonal and inter-annual roles of lakes in landscape and global carbon cycling, particularly in a global change context. Finally, we highlighted critical data needs, which include surface water DOC observations in paired tributary and lake systems, measurements of OC burial rates, groundwater input volume and DOC, and budgets of particulate OC.
US National Science Foundation's MacroSystems Biology Program [EF1137353, EF1137327]; NSF Long-Term Ecological Research program (North Temperate Lakes) [DEB-1440297]
This project was a product of the Global Lake Ecological Observatory Network (GLEON) Fellowship program supported by the US National Science Foundation's MacroSystems Biology Program (Awards #EF1137353 and EF1137327). Logistical support was provided by the University of Wisconsin-Madison Center for Limnology, the Cary Institute of Ecosystem Studies, the University of Wisconsin Trout Lake Station, the Lake Sunapee Protective Association, and Grace Hong. Limnological data providers included the NSF Long-Term Ecological Research program (North Temperate Lakes DEB-1440297 and Arctic LTER), the Swedish Meteorological and Hydrological Institute, the Swedish University of Agricultural Sciences and the Canadian Dorset Environmental Science Centre. Additional details on data sources were included online in the appendix (Supplementary material). IMM, HAD, KJF, AMM, ZO, DR, FS and PCH acquired data, developed the model, and performed data analyses. All authors participated in conceiving and developing the project and writing the paper. We thank two reviewers for constructive comments on a draft of this manuscript. Analysis scripts and public data (LTER lakes and Lake Vanern) can be freely downloaded here: https://github.com/GLEON/SOS.
application/pdf
en
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
Carbon cycle
Mass balance
Dissolved organic carbon
Particulate organic carbon
LTER
GLEON
Dynamic modeling of organic carbon fates in lake ecosystems
Ecological Modelling
Article - Refereed
Text
StillImage
ORIGINAL
1-s2.0-S0304380018302783-main.pdf
1-s2.0-S0304380018302783-main.pdf
application/pdf
5342964
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/6e9b88ba-d452-4c22-a652-3340e0597411/download
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MD5
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1-s2.0-S0304380018302783-main.pdf.txt
1-s2.0-S0304380018302783-main.pdf.txt
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70294
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MD5
5
10919/93392
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/93392
2022-02-26 15:58:35.69
http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/468362020-10-28T05:42:01Zcom_10919_23829com_10919_5553col_10919_23830
Virginia Tech
Stevens, Glen N.
Jones, R. H.
Biological Sciences
2014-03-27T13:06:04Z
2014-03-27T13:06:04Z
2006-03
2014-03-11
Glen N. Stevens and Robert H. Jones 2006. PATTERNS IN SOIL FERTILITY AND ROOT HERBIVORY INTERACT TO INFLUENCE FINE-ROOT DYNAMICS. Ecology 87:616-624. http://dx.doi.org/10.1890/05-0809
0012-9658
http://hdl.handle.net/10919/46836
http://www.esajournals.org/doi/pdf/10.1890/05-0809
https://doi.org/10.1890/05-0809
Fine-scale soil nutrient enrichment typically stimulates root growth, but it may also increase root herbivory, resulting in trade-offs for plant species and potentially influencing carbon cycling patterns. We used root ingrowth cores to investigate the effects of microsite fertility and root herbivory on root biomass in an aggrading upland forest in the coastal plain of South Carolina, USA. Treatments were randomly assigned to cores from a factorial combination of fertilizer and insecticide. Soil, soil fauna, and roots were removed from the cores at the end of the experiment (8-9 mo), and roots were separated at harvest into three diameter classes. Each diameter class responded differently to fertilizer and insecticide treatments. The finest roots (< 1.0 mm diameter), which comprised well over half of all root biomass, were the only ones to respond significantly to both treatments, increasing when fertilizer and when insecticide were added (each P < 0.0001), with maximum biomass found where the treatments were combined (interaction term significant, P < 0.001). These results Suggest that root-feeding insects have a strong influence on root standing crop with stronger herbivore impacts on finer roots and within more fertile microsites. Thus, increased vulnerability to root herbivory is a potentially significant,cost of root foraging in nutrient-rich patches.
National Science Foundation Grant DEB-0308847
Virginia Tech Graduate Student Assembly
Virginia Tech Department of Biological Sciences
en_US
Ecological Society of America
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Ecological Society of America
elateridae
heterogeneity
root foraging
root ingrowth core
scarabaeidae
early plant succession
bottom-up forces
insect herbivory
top-down
nutrient heterogeneity
community structure
tallgrass prairie
spatial
variation
united-states
responses
Patterns in soil fertility and root herbivory interact to influence fine-root dynamics
Ecology
Article - Refereed
ORIGINAL
05-0809.pdf
05-0809.pdf
Main article
application/pdf
119489
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05-0809.pdf.txt
05-0809.pdf.txt
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45918
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MD5
4
10919/46836
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/46836
2020-10-28 05:42:01.309
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/851882023-04-14T17:49:45Zcom_10919_23829com_10919_5553col_10919_23830
Gross, Fridolin
Bonaiuti, Paolo
Hauf, Silke
Ciliberto, Andrea
Biological Sciences
Fralin Life Sciences Institute
2018-09-28T14:02:02Z
2018-09-28T14:02:02Z
2018-09-10
e1006449
http://hdl.handle.net/10919/85188
https://doi.org/10.1371/journal.pcbi.1006449
14
9
The mitotic checkpoint (also called spindle assembly checkpoint) is a signaling pathway that ensures faithful chromosome segregation. Mitotic checkpoint proteins inhibit the anaphase-promoting complex (APC/C) and its activator Cdc20 to prevent precocious anaphase. Checkpoint signaling leads to a complex of APC/C, Cdc20, and checkpoint proteins, in which the APC/C is inactive. In principle, this final product of the mitotic checkpoint can be obtained via different pathways, whose relevance still needs to be fully ascertained experimentally. Here, we use mathematical models to compare the implications on checkpoint response of the possible pathways leading to APC/C inhibition. We identify a previously unrecognized funneling effect for Cdc20, which favors Cdc20 incorporation into the inhibitory complex and therefore promotes checkpoint activity. Furthermore, we find that the presence or absence of one specific assembly reaction determines whether the checkpoint remains functional at elevated levels of Cdc20, which can occur in cancer cells. Our results reveal the inhibitory logics behind checkpoint activity, predict checkpoint efficiency in perturbed situations, and could inform molecular strategies to treat malignancies that exhibit Cdc20 overexpression.
Published version
application/pdf
en
PLOS
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Implications of alternative routes to APC/C inhibition by the mitotic checkpoint complex
PLOS Computational Biology
Article - Refereed
Text
ORIGINAL
journal.pcbi.1006449.pdf
journal.pcbi.1006449.pdf
application/pdf
2558313
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MD5
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journal.pcbi.1006449.pdf.txt
journal.pcbi.1006449.pdf.txt
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65668
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965e29675cd5d678457827d0f9395de6
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MD5
5
10919/85188
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/85188
2023-04-14 17:49:45.508
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1010272022-02-26T22:17:37Zcom_10919_23829com_10919_5553col_10919_23830
De Rienzo, Assunta
Chirieac, Lucian R.
Hung, Yin P.
Severson, David T.
Freyaldenhoven, Samuel
Gustafson, Corinne E.
Dao, Nhien T.
Meyerovitz, Claire, V
Oster, Michela E.
Jensen, Roderick V.
Yeap, Beow Y.
Bueno, Raphael
Richards, William G.
Biological Sciences
2020-12-07T14:12:49Z
2020-12-07T14:12:49Z
2020-09-17
0022-3417
http://hdl.handle.net/10919/101027
https://doi.org/10.1002/path.5551
32944962
1096-9896
BRCA1-associated protein-1(BAP1) expression is commonly lost in several tumors including malignant pleural mesothelioma (MPM). Presence or absence of immunohistochemical BAP1 nuclear staining in tumor cells is currently used for differential diagnosis of MPM. In this study, a large cohort of 596 MPM tumors with available clinical data was analyzed to examine associations of BAP1 staining pattern with clinical and molecular features that may reflect the impact ofBAP1mutation on MPM biology. Cases were classified according to the BAP1 staining pattern of tumor cells. Exome and RNA-sequencing data were available for subsets of cases. Levels of mRNA encoding claudin 15 (CLDN15) and vimentin (VIM) were determined using RT-qPCR on 483 cases to estimate the relative proportions of epithelial-like and mesenchymal-like components in each tumor. Four BAP1 staining patterns were observed: single-pattern nuclear staining (36%), single-pattern cytoplasmic staining (25%), single-pattern absent staining (12%), and combinations of these staining patterns (27%). This study confirmed prior reports that nuclear BAP1 is more frequently associated with wild-typeBAP1and sarcomatoid histology. However, no associations between BAP1 staining pattern(s) and mutations in specific protein domains and/or mutation type were observed. BAP1 staining patterns were significantly associated (p < 0.001) withBAP1gene expression, MPM histologic subtypes, molecular clusters, and markers of epithelial-to-mesenchymal transition. Frequent observation of combinations of BAP1 staining patterns in MPM tumors indicated intra-tumoral heterogeneity ofBAP1status. Cytoplasmic BAP1 staining was identified as a putative indicator of favorable prognosis in non-epithelioid MPM. In conclusion, novel significant associations among different BAP1 staining patterns and subgroups of MPM tumors were observed, suggesting that the role ofBAP1in tumor progression may be more complex than its presumed tumor suppressor function. Cytoplasmic staining was identified as a putative indicator of favorable prognosis in non-epithelioid MPM, potentially addressing a critical need in clinical decision-making in this disease. (c) 2020 The Authors.The Journal of Pathologypublished by John Wiley & Sons, Ltd. on behalf of The Pathological Society of Great Britain and Ireland.
National Cancer InstituteUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Cancer Institute (NCI) [NCI 2 R01 CA120528-11A1]; International Mesothelioma Program at Brigham and Women's Hospital; NCI Cancer Center Support GrantUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Cancer Institute (NCI) [NIH 5 P30 CA06516]
This work was supported by grants to RB from the National Cancer Institute (NCI 2 R01 CA120528-11A1) and the International Mesothelioma Program at Brigham and Women's Hospital. The study sponsors played no role in the study design, collection, analysis, interpretation of data, writing of the report, or decision to submit the paper for publication. We thank the Dana-Farber/Harvard Cancer Center in Boston, MA, for the use of the Specialized Histopathology Core, which provided histology and immunohistochemistry service. Dana-Farber/Harvard Cancer Center is supported in part by an NCI Cancer Center Support Grant #NIH 5 P30 CA06516.
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
mesothelioma
BAP1
immunohistochemistry
tumor suppressor gene
intra-tumor heterogeneity
epithelial-to-mesenchymal transition
gene expression
prognostic biomarker
Large-scale analysis of BAP1 expression reveals novel associations with clinical and molecular features of malignant pleural mesothelioma
Journal of Pathology
Article - Refereed
Text
StillImage
TEXT
path.5551.pdf.txt
path.5551.pdf.txt
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text/plain
56229
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f3210b213d28bbf24b768c12f4fe8ea6
MD5
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MD5
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path.5551.pdf
path.5551.pdf
application/pdf
11851746
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MD5
1
10919/101027
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/101027
2022-02-26 22:17:37.89
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1094772023-11-29T19:06:54Zcom_10919_5540com_10919_23829com_10919_5553col_10919_71752col_10919_23830
Kaler, Kylie M. Ryan
Nix, Jay C.
Schubot, Florian D.
2022-03-29T12:17:00Z
2022-03-29T12:17:00Z
2021-10
101193
http://hdl.handle.net/10919/109477
https://doi.org/10.1016/j.jbc.2021.101193
297
4
34529974
1083-351X
Bacterial signaling histidine kinases (HKs) have long been postulated to function exclusively through linear signal transduction chains. However, several HKs have recently been shown to form complex multikinase networks (MKNs). The most prominent MKN, involving the enzymes RetS and GacS, controls the switch between the motile and biofilm lifestyles in the pathogenic bacterium Pseudomonas aeruginosa. While GacS promotes biofilm formation, RetS counteracts GacS using three distinct mechanisms. Two are dephosphorylating mechanisms. The third, a direct binding between the RetS and GacS HK regions, blocks GacS autophosphorylation. Focusing on the third mechanism, we determined the crystal structure of a cocomplex between the HK region of RetS and the dimerization and histidine phosphotransfer (DHp) domain of GacS. This is the first reported structure of a complex between two distinct bacterial signaling HKs. In the complex, the canonical HK homodimerization interface is replaced by a strikingly similar heterodimeric interface between RetS and GacS. We further demonstrate that GacS autophosphorylates in trans, thus explaining why the formation of a RetS-GacS complex inhibits GacS autophosphorylation. Using mutational analysis in conjunction with bacterial two-hybrid and biofilm assays, we not only corroborate the biological role of the observed RetSGacS interactions, but also identify a residue critical for the equilibrium between the RetS-GacS complex and the respective RetS and GacS homodimers. Collectively, our findings suggest that RetS and GacS form a domain-swapped hetero-oligomer during the planktonic growth phase of P. aeruginosa before unknown signals cause its dissociation and a relief of GacS inhibition to promote biofilm formation.
NIHUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USA [R21AI128255-01A1]
Published version
This study was supported by NIH grant R21AI128255-01A1 to F. D. S. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
application/pdf
en
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
RetS inhibits Pseudomonas aeruginosa biofilm formation by disrupting the canonical histidine kinase dimerization interface of GacS
Journal of Biological Chemistry
Article - Refereed
Text
ORIGINAL
PIIS0021925821009959.pdf
PIIS0021925821009959.pdf
Published version
application/pdf
2765653
https://vtechworks.lib.vt.edu/bitstreams/796c4c62-9c0b-47d3-9ec2-8a5418f7b388/download
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MD5
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PIIS0021925821009959.pdf.txt
PIIS0021925821009959.pdf.txt
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59760
https://vtechworks.lib.vt.edu/bitstreams/d1e5da5c-7a26-4e4c-8883-7bd5d9be38bb/download
81b222ebc92897f4185e9c5c7d18b292
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MD5
4
10919/109477
oai:vtechworks.lib.vt.edu:10919/109477
2023-11-29 19:06:54.76
http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1023812021-02-17T12:51:52Zcom_10919_78629com_10919_78628com_10919_23829com_10919_5553com_10919_91913com_10919_23198col_10919_78791col_10919_23830col_10919_91914
Motahari, Zahra
Maynard, Thomas M.
Popratiloff, Anastas
Moody, Sally A.
LaMantia, Anthony-Samuel
Fralin Biomedical Research Institute
Biological Sciences
2021-02-16T13:49:46Z
2021-02-16T13:49:46Z
2020-09-15
0964-6906
http://hdl.handle.net/10919/102381
https://doi.org/10.1093/hmg/ddaa199
29
18
32901287
1460-2083
We identified divergent modes of initial axon growth that prefigure disrupted differentiation of the trigeminal nerve (CN V), a cranial nerve essential for suckling, feeding and swallowing (S/F/S), a key innate behavior compromised in multiple genetic developmental disorders including DiGeorge/22q11.2 Deletion Syndrome (22q11.2 DS). We combined rapid in vivo labeling of single CN V axons in LgDel(+/-) mouse embryos, a genomically accurate 22q11.2DS model, and 3D imaging to identify and quantify phenotypes that could not be resolved using existing methods. We assessed these phenotypes in three 22q11.2-related genotypes to determine whether individual CN V motor and sensory axons wander, branch and sprout aberrantly in register with altered anterior-posterior hindbrain patterning and gross morphological disruption of CN V seen in LgDel(+/-). In the additional 22q11.2-related genotypes: Tbx1(+/-), Ranbp1(+/-), Ranbp1(+/-) and LgDel(+/-):Raldh2(+/-); axon phenotypes are seen when hindbrain patterning and CN V gross morphology is altered, but not when it is normal or restored toward WT. This disordered growth of CN V sensory and motor axons, whose appropriate targeting is critical for optimal S/F/S, may be an early, critical determinant of imprecise innervation leading to inefficient oropharyngeal function associated with 22q11.2 deletion from birth onward.
National Institute of Child Health and Human DevelopmentUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD) [P01 HD083157]; NICHD IDDRC U54 [HD 090257]; Georgetown University; Children's National Medical Center
National Institute of Child Health and Human Development (P01 HD083157 to A.S.L.); NICHD IDDRC U54 (HD 090257 to G.W.); Georgetown University and Children's National Medical Center.
application/pdf
en
Creative Commons Attribution-NonCommercial 4.0 International
http://creativecommons.org/licenses/by-nc/4.0/
Aberrant early growth of individual trigeminal sensory and motor axons in a series of mouse genetic models of 22q11.2 deletion syndrome
Human Molecular Genetics
Article - Refereed
Text
StillImage
ORIGINAL
ddaa199.pdf
ddaa199.pdf
application/pdf
2391826
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/7449e2e9-f1d6-4850-ac95-4fd7821873e5/download
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MD5
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ddaa199.pdf.txt
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73210
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d154363d36b98da6ef8c7fac3bea7dac
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MD5
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10919/102381
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/102381
2021-02-17 12:51:52.09
http://creativecommons.org/licenses/by-nc/4.0/
Creative Commons Attribution-NonCommercial 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/897412020-10-22T03:35:27Zcom_10919_23829com_10919_5553col_10919_23830
Opell, Brent D.
Clouse, Mary E.
Andrews, Sheree F.
Biological Sciences
2019-06-04T16:57:38Z
2019-06-04T16:57:38Z
2018-05-30
e0196972
http://hdl.handle.net/10919/89741
https://doi.org/10.1371/journal.pone.0196972
13
5
29847578
1932-6203
An orb web's prey capture thread features tiny glue droplets, each formed of an adhesive glycoprotein core surrounded by an aqueous layer. Small molecules in the aqueous layer confer droplet hygroscopicity and maintain glycoprotein viscoelasticity, causing droplet volume and glycoprotein performance to track changes in environmental humidity. Droplet extension combines with that of a thread's supporting flagelliform fibers to sum the adhesive forces of multiple droplets, creating an effective adhesive system. We combined measurements of the force on an extending droplet, as gauged by the deflection of its support line, with measurements of glycoprotein volume and droplet extension to determine the Young's modulus (E) and toughness of three species' glycoproteins. We did this at five relative humidities between 20 - 90% to assess the effect of humidity on these properties. When droplets of a thread span extend, their extensions are constrained and their glycoprotein filaments remain covered by aqueous material. This was also the case during the first extension phase of the individual droplets that we examined. However, as extension progressed, the aqueous layer was progresses disrupted, exposing the glycoprotein. During the first extension phase E ranged from 0.00003 GPa, a value similar to that of fibronectin, a glycoprotein that anchors cells in the extracellular matrix, to 0.00292 GPa, a value similar to that of resilin in insect ligaments. Second phase E increased 4.7 - 19.4-fold. When compared at the same humidity the E of each species' glycoprotein was less than 5% of the value reported for its flagelliform fibers. This difference may facilitate the coordinated extension of these two capture thread components that is responsible for summing the thread's adhesive forces.
National Science Foundation grant [IOS-1257719]
National Science Foundation grant IOS-1257719 (BO) supported this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
application/pdf
en
PLOS
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Elastic modulus and toughness of orb spider glycoprotein glue
PLOS ONE
Article - Refereed
Text
StillImage
ORIGINAL
journal.pone.0196972.pdf
journal.pone.0196972.pdf
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10919/89741
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/89741
2020-10-22 03:35:27.354
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1177822024-02-01T10:00:31Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553col_10919_70873col_10919_23830
Sonani, Ravi R.
Palmer, Lee K.
Esteves, Nathaniel C.
Horton, Abigail A.
Sebastian, Amanda L.
Kelly, Rebecca J.
Wang, Fengbin
Kreutzberger, Mark A. B.
Russell, William K.
Leiman, Petr G.
Scharf, Birgit E.
Egelman, Edward H.
2024-02-01T14:11:08Z
2024-02-01T14:11:08Z
2024-01-26
756 (Article number)
2041-1723
10.1038/s41467-024-44959-z (PII)
https://hdl.handle.net/10919/117782
https://doi.org/10.1038/s41467-024-44959-z
15
1
Scharf, Birgit [0000-0001-6271-8972]
38272938
2041-1723
A contractile sheath and rigid tube assembly is a widespread apparatus used by bacteriophages, tailocins, and the bacterial type VI secretion system to penetrate cell membranes. In this mechanism, contraction of an external sheath powers the motion of an inner tube through the membrane. The structure, energetics, and mechanism of the machinery imply rigidity and straightness. The contractile tail of Agrobacterium tumefaciens bacteriophage Milano is flexible and bent to varying degrees, which sets it apart from other contractile tail-like systems. Here, we report structures of the Milano tail including the sheath-tube complex, baseplate, and putative receptor-binding proteins. The flexible-to-rigid transformation of the Milano tail upon contraction can be explained by unique electrostatic properties of the tail tube and sheath. All components of the Milano tail, including sheath subunits, are crosslinked by disulfides, some of which must be reduced for contraction to occur. The putative receptor-binding complex of Milano contains a tailspike, a tail fiber, and at least two small proteins that form a garland around the distal ends of the tailspikes and tail fibers. Despite being flagellotropic, Milano lacks thread-like tail filaments that can wrap around the flagellum, and is thus likely to employ a different binding mechanism.
Published version
application/pdf
en
Springer
https://www.ncbi.nlm.nih.gov/pubmed/38272938
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
An extensive disulfide bond network prevents tail contraction in Agrobacterium tumefaciens phage Milano
Nature Communications
Article - Refereed
Journal Article
Text
2024-01-10
/Virginia Tech
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/All T&R Faculty
/Virginia Tech/Science/COS T&R Faculty
LICENSE
license.txt
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s41467-024-44959-z.pdf
Published version
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s41467-024-44959-z.pdf.txt
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10919/117782
oai:vtechworks.lib.vt.edu:10919/117782
2024-02-01 10:00:31.363
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
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
oai:vtechworks.lib.vt.edu:10919/249722022-03-29T18:37:35Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:22Z
2014-01-23T02:36:22Z
2005
http://hdl.handle.net/10919/24972
http://www.johncairns.net/Papers/PlanetarySustainability.pdf
Arguably, no curve in the world increases indefinitely--- certainly no growth curve of human society. The quest for sustainable use of the planet involves estimating levels of activity, particularly resource use, that can continue indefinitely. Since the biosphere is dynamic, this continuation is not a stagnant, steady-state situation but rather a mutualistic interactive relationship between human society and the planet's ecological life support system. Human technology, creativity, and ingenuity may modify natural laws, but cannot be used to repeal them. Attempts to maintain the recent exponential growth of the human population, affluence, and artifacts cannot continue forever, or probably even for another century. At best, quality of life will be diminished, and, at worst, a substantial loss of human life will occur if both ethical and ecological issues are not freely and openly discuessed. Sustainability may be visionary and unattainable, but it does offer an opportunity to improve the quality of life for futrure generations of humans and other life forms, an opportunity that is far greater than possible with present unsustainable practices.
en_US
Association for Politics and the Life Sciences
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Association for Politics and the Life Sciences
Sustainability
resource use
biosphere
exponential growth period
mass migration
ecological footprint
precautionary principle
Planetary Sustainability the Age of Transition to Sustainablility: The End of the Exponential Growth Period
Article
Text
ORIGINAL
cairns_planetary_sustainability.pdf
application/pdf
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cairns_planetary_sustainability.pdf.txt
cairns_planetary_sustainability.pdf.txt
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10919/24972
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/24972
2022-03-29 18:37:35.921
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1095722022-04-07T07:12:12Zcom_10919_78629com_10919_78628com_10919_23829com_10919_5553col_10919_78630col_10919_23830
Zipper, Samuel C.
Hammond, John C.
Shanafield, Margaret
Zimmer, Margaret
Datry, Thibault
Jones, C. Nathan
Kaiser, Kendra E.
Godsey, Sarah E.
Burrows, Ryan M.
Blaszczak, Joanna R.
Busch, Michelle H.
Price, Adam N.
Boersma, Kate S.
Ward, Adam S.
Costigan, Katie
Allen, George H.
Krabbenhoft, Corey A.
Dodds, Walter K.
Mims, Meryl C.
Olden, Julian D.
Kampf, Stephanie K.
Burgin, Amy J.
Allen, Daniel C.
2022-04-06T14:34:57Z
2022-04-06T14:34:57Z
2021-08
1748-9326
84033
http://hdl.handle.net/10919/109572
https://doi.org/10.1088/1748-9326/ac14ec
16
8
Non-perennial streams are widespread, critical to ecosystems and society, and the subject of ongoing policy debate. Prior large-scale research on stream intermittency has been based on long-term averages, generally using annually aggregated data to characterize a highly variable process. As a result, it is not well understood if, how, or why the hydrology of non-perennial streams is changing. Here, we investigate trends and drivers of three intermittency signatures that describe the duration, timing, and dry-down period of stream intermittency across the continental United States (CONUS). Half of gages exhibited a significant trend through time in at least one of the three intermittency signatures, and changes in no-flow duration were most pervasive (41% of gages). Changes in intermittency were substantial for many streams, and 7% of gages exhibited changes in annual no-flow duration exceeding 100 days during the study period. Distinct regional patterns of change were evident, with widespread drying in southern CONUS and wetting in northern CONUS. These patterns are correlated with changes in aridity, though drivers of spatiotemporal variability were diverse across the three intermittency signatures. While the no-flow timing and duration were strongly related to climate, dry-down period was most strongly related to watershed land use and physiography. Our results indicate that non-perennial conditions are increasing in prevalence over much of CONUS and binary classifications of 'perennial' and 'non-perennial' are not an accurate reflection of this change. Water management and policy should reflect the changing nature and diverse drivers of changing intermittency both today and in the future.
US National Science FoundationNational Science Foundation (NSF) [DEB-1754389]
Published version
This manuscript is a product of the Dry Rivers Research Coordination Network, which was supported by funding from the US National Science Foundation (DEB-1754389). Any use of trade, firm, or product names is for descriptive purposes only and does not imply endorsement by the US Government. This manuscript was improved by constructive feedback from Kristin Jaeger and three anonymous reviews.
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
non-perennial streams
climate change
land use
river
streamflow
ephemeral
time series
Pervasive changes in stream intermittency across the United States
Environmental Research Letters
Article - Refereed
Text
ORIGINAL
Zipper_2021_Environ._Res._Lett._16_084033.pdf
Zipper_2021_Environ._Res._Lett._16_084033.pdf
Published version
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4433841
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Zipper_2021_Environ._Res._Lett._16_084033.pdf.txt
Zipper_2021_Environ._Res._Lett._16_084033.pdf.txt
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68575
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10919/109572
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/109572
2022-04-07 07:12:12.188
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/751972023-11-29T19:07:00Zcom_10919_5540com_10919_23829com_10919_5553col_10919_71752col_10919_23830
Li, Chengyun
Wallace, Regina A.
Black, Wesley P.
Li, Yue-zhong
Yang, Zhaomin
Biological Sciences
2017-02-28T19:20:02Z
2017-02-28T19:20:02Z
2013-07-26
http://hdl.handle.net/10919/75197
https://doi.org/10.1371/journal.pone.0070144
8
7
The bacterial type IV pilus (T4P) is the strongest biological motor known to date as its retraction can generate forces well over 100 pN. Myxococcus xanthus, a 𝞭-proteobacterium, provides a good model for T4P investigations because its social (S) gliding motility is powered by T4P. In this study, the interactions among M. xanthus T4P proteins were investigated using genetics and the yeast two-hybrid (Y2H) system. Our genetic analysis suggests that there is an integrated T4P structure that crosses the inner membrane (IM), periplasm and the outer membrane (OM). Moreover, this structure exists in the absence of the pilus filament. A systematic Y2H survey provided evidence for direct interactions among IM and OM proteins exposed to the periplasm. For example, the IM lipoprotein PilP interacted with its cognate OM protein PilQ. In addition, interactions among T4P proteins from the thermophile Thermus thermophilus were investigated by Y2H. The results indicated similar protein-protein interactions in the T4P system of this non-proteobacterium despite significant sequence divergence between T4P proteins in T. thermophilus and M. xanthus. The observations here support the model of an integrated T4P structure in the absence of a pilus in diverse bacterial species.
application/pdf
en_US
PLoS
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Type IV Pilus Proteins Form an Integrated Structure Extending from the Cytoplasm to the Outer Membrane
PLoS ONE
1932-6203
600
Article - Refereed
Text
ORIGINAL
YangTypeIV.pdf
YangTypeIV.pdf
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LICENSE
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license.txt
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YangTypeIV.pdf.txt
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10919/75197
oai:vtechworks.lib.vt.edu:10919/75197
2023-11-29 19:07:00.09
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
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oai:vtechworks.lib.vt.edu:10919/257702020-11-24T19:19:53Zcom_10919_23829com_10919_5553col_10919_23830
Virginia Tech
Gerard, Claude
Tyson, John J.
Novak, Bela
Biological Sciences
2014-02-26T19:10:04Z
2014-02-26T19:10:04Z
2013-03
2014-02-05
Gerard, Claude; Tyson, John J.; Novak, Bela. "Minimal Models for Cell-Cycle Control Based on Competitive Inhibition and Multisite Phosphorylations of Cdk Substrates," Biophysical Journal 104(6), 1367-1379 (2013); doi: 10.1016/j.bpj.2013.02.012
0006-3495
http://hdl.handle.net/10919/25770
http://www.sciencedirect.com/science/article/pii/S0006349513002026
https://doi.org/10.1016/j.bpj.2013.02.012
The eukaryotic cell cycle is characterized by alternating oscillations in the activities of cyclin-dependent kinase (Cdk) and the anaphase-promoting complex (APC). Successful completion of the cell cycle is dependent on the precise, temporally ordered appearance of these activities. A modest level of Cdk activity is sufficient to initiate DNA replication, but mitosis and APC activation require an elevated Cdk activity. In present-day eukaryotes, this temporal order is provided by a complex network of regulatory proteins that control both Cdk and APC activities via sharp thresholds, bistability, and time delays. Using simple computational models, we show here that these dynamical features of cell-cycle organization could emerge in a control system driven by a single Cdk/cyclin complex and APC wired in a negative-feedback loop. We show that ordered phosphorylation of cellular proteins could be explained by multisite phosphorylation/dephosphorylation and competition of substrates for interconverting kinase (Cdk) and phosphatase. In addition, the competition of APC substrates for ubiquitylation can create and maintain sustained oscillations in cyclin levels. We propose a sequence of models that gets closer and closer to a realistic model of cell-cycle control in yeast. Since these models lack the elaborate control mechanisms characteristic of modern eukaryotes, they suggest that bistability and time delay may have characterized eukaryotic cell divisions before the current cell-cycle control network evolved in all its complexity.
National Institutes of Health 5R01-GM078989-07, 1U54-CA149147-03
Biotechnology and Biological Sciences Research Council
European Community's Seventh Framework Programmes UniCellSys/201142, MitoSys/241548
en_US
CELL PRESS
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Biophysical Society
quantitative model
fission yeast
mitotic exit
s-phase
kinase
network
cooperativity
bistability
dynamics
securin
Minimal Models for Cell-Cycle Control Based on Competitive Inhibition and Multisite Phosphorylations of Cdk Substrates
Biophysical Journal
Article - Refereed
ORIGINAL
1-s2.0-S0006349513002026-main.pdf
1-s2.0-S0006349513002026-main.pdf
Main article
application/pdf
5513244
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MD5
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MD5
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10919/25770
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/25770
2020-11-24 19:19:53.742
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/468282021-01-08T13:38:48Zcom_10919_23829com_10919_5553col_10919_23830
Virginia Tech
Nilsen, Erik T.
Sharifi, M. Rasoul
Rundel, Philip W.
Biological Sciences
2014-03-27T13:06:02Z
2014-03-27T13:06:02Z
1984
2014-03-11
Erik Tallak Nilsen, M. Rasoul Sharifi, and Philip W. Rundel 1984. Comparative Water Relations of Phreatophytes in the Sonoran Desert of California. Ecology 65:767-778. http://dx.doi.org/10.2307/1938049
0012-9658
http://hdl.handle.net/10919/46828
http://www.jstor.org/stable/1938049
https://doi.org/10.2307/1938049
The seasonal and diurnal water relations were compared among six desert phreatophytes, two evergreen shrubs, and one deciduous shrub. All species were located in one wash woodland in the Sonoran Desert of southern California. There are several mechanisms by which these Phaenicia have adapted to the desert environment. One group of winter-deciduous phreatophytes (Olneya tesota, Prosopis glandulosa, and Acacia greggii) experienced summer midday leaf water potentials below -4.0 MPa. These phreatophytes had a series of physiological mechanisms for tolerating summer water stress, including seasonal and diurnal osmotic adjustment and the maintenance of high leaf conductance at low leaf water potential. Osmotic adjustment of these three phreatophytes was similar to or greater than that of two evergreen species (Larrea tridentata and Simmondsia chinensis). Dalea spinosa, a stem-photosynthetic phreatophyte, avoided water stress by maintaining a very small leaf area. The summer-deciduous phreatophytes (Hyptis emoryi, and Chilopsis linearis) demonstrated mechanisms of drought avoidance such as change in leaf biomass and low summer leaf conductance. Little osmotic adjustment occurred in the summer-deciduous phreatophytes. The phreatophytic species studied in this investigation have evolved adaptations to water stress that are similar to those of deciduous and evergreen shrubs of the Sonoran Desert. Desert phreatophytes are a complex group of species with varied adaptive mechanisms to tolerate or avoid drought and should not be considered simply as a group of species that avoid desert water stress by utilizing deep ground water unavailable to other desert species of drought tolerance and avoidance.
NSF DEB 79-21971
en_US
Ecological Society of America
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Ecological Society of America
Acacia greggii
Chilopsis linearis
Dalea spinosa
Hyptis emoryi
leaf conductance
Olneya tesota
osmotic adjustment
phreatophytes
Prosopis glandulosa
Sonoran Desert
water relations
Comparative water relations of phreatophytes in the sonoran desert of california
Ecology
Article - Refereed
ORIGINAL
1938049.pdf
1938049.pdf
Main article
application/pdf
1934028
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/cd04d63e-9e86-415c-a29b-bf109f6792f5/download
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MD5
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TEXT
1938049.pdf.txt
1938049.pdf.txt
Extracted text
text/plain
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MD5
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10919/46828
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/46828
2021-01-08 13:38:48.724
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1067072023-12-19T13:34:26Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553com_10919_24259com_10919_5559com_10919_91913com_10919_23198com_10919_91912col_10919_70873col_10919_23830col_10919_24342col_10919_91914col_10919_91916
Rai, Pallavi
Chuong, Christina
LeRoith, Tanya
Smyth, James W.
Panov, Julia
Levi, Moshe
Kehn-Hall, Kylene
Duggal, Nisha K.
Weger-Lucarelli, James
2021-11-22T17:41:00Z
2021-11-22T17:41:00Z
2021-11-01
2021-11-22T17:40:52Z
0042-6822
S0042-6822(21)00184-7 (PII)
http://hdl.handle.net/10919/106707
https://doi.org/10.1016/j.virol.2021.08.014
563
Duggal, Nisha [0000-0002-9893-8069]
Kehn-Hall, Kylene [0000-0001-8036-7213]
LeRoith, Tanya [0000-0002-1196-6949]
34509029
1089-862X
The COVID-19 pandemic has paralyzed the global economy and resulted in millions of deaths globally. People with co-morbidities like obesity, diabetes and hypertension are at an increased risk for severe COVID-19 illness. This is of overwhelming concern because 42% of Americans are obese, 30% are pre-diabetic and 9.4% have clinical diabetes. Here, we investigated the effect of obesity on disease severity following SARS-CoV-2 infection using a well-established mouse model of diet-induced obesity. Diet-induced obese and lean control C57BL/6 N mice, transduced for ACE2 expression using replication-defective adenovirus, were infected with SARS-CoV-2, and monitored for lung pathology, viral titers, and cytokine expression. No significant differences in tissue pathology or viral replication was observed between AdV transduced lean and obese groups, infected with SARS-CoV-2, but certain cytokines were expressed more significantly in infected obese mice compared to the lean ones. Notably, significant weight loss was observed in obese mice treated with the adenovirus vector, independent of SARS-CoV-2 infection, suggesting an obesity-dependent morbidity induced by the vector. These data indicate that the adenovirus-transduced mouse model of SARS-CoV-2 infection, as described here and elsewhere, may be inappropriate for nutrition studies.
Published version
Pages 98-106
9 page(s)
application/pdf
en
Elsevier
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000700278300004&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Life Sciences & Biomedicine
Virology
COVID-19
Diet-induced obesity
hACE2
Replication-defective adenovirus
Tissue
Pathology
ACUTE RESPIRATORY SYNDROME
SYNDROME CORONAVIRUS INFECTION
IMMUNE-RESPONSES
MORTALITY
SARS
Virology
06 Biological Sciences
07 Agricultural and Veterinary Sciences
11 Medical and Health Sciences
Vero Cells
Animals
Mice, Inbred C57BL
Humans
Mice
Obesity
Disease Models, Animal
Morbidity
Comorbidity
Female
Male
HEK293 Cells
Chlorocebus aethiops
COVID-19
Adenovirus transduction to express human ACE2 causes obesity-specific morbidity in mice, impeding studies on the effect of host nutritional status on SARS-CoV-2 pathogenesis
Virology
Article - Refereed
Article
Journal
Text
2021-08-31
/Virginia Tech
/Virginia Tech/Veterinary Medicine
/Virginia Tech/Veterinary Medicine/Biomedical Sciences and Pathobiology
/Virginia Tech/University Research Institutes
/Virginia Tech/University Research Institutes/Fralin Life Sciences
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/All T&R Faculty
/Virginia Tech/Veterinary Medicine/CVM T&R Faculty
/Virginia Tech/University Research Institutes/Fralin Life Sciences/Durelle Scott
SWORD
sword-2021-11-22T12:40:52.original.xml
Original SWORD entry document
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Adenovirus transduction to express human ACE2 causes obesity-specific morbidity in mice, impeding studies on the effect of h.pdf.txt
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ORIGINAL
Adenovirus transduction to express human ACE2 causes obesity-specific morbidity in mice, impeding studies on the effect of h.pdf
Adenovirus transduction to express human ACE2 causes obesity-specific morbidity in mice, impeding studies on the effect of h.pdf
Published version
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10919/106707
oai:vtechworks.lib.vt.edu:10919/106707
2023-12-19 13:34:26.788
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1046472023-11-29T19:07:02Zcom_10919_5540com_10919_23829com_10919_5553com_10919_23965com_10919_5557col_10919_71752col_10919_23830col_10919_24280
Sleezer, Logan J.
Angermeier, Paul L.
Frimpong, Emmanuel A.
Brown, Bryan L.
Fish and Wildlife Conservation
Biological Sciences
2021-08-16T17:31:12Z
2021-08-16T17:31:12Z
2021-08-10
http://hdl.handle.net/10919/104647
https://doi.org/10.1111/ddi.13393
Aim
We developed a new technique, utilizing species-specific counts of individuals from historical fish community samples, to examine landscape-level, spatio-temporal trends in relative abundance distributions. Abundance-based historical distribution analyses are often plagued by data comparability issues, but provide critical information about community composition trends inaccessible to those using analyses based only on species presence–absence. We established trends in native and non-native fish abundance and community homogenization, uniqueness and diversity to help local conservation managers prioritize targets and motivate similar studies globally to support fish conservation.
Location
Upper and middle New River (UMNR) basin, Appalachian Mountains, USA.
Methods
We compiled catch data from 61 years of fish community surveys (1958–2019) and tested for community homogenization by comparing data from repeatedly sampled sites (1900s versus 2000s samples) using dispersion analyses. We measured community uniqueness (site contributions to beta diversity) and species diversity (Shannon index) at sampled streams to identify potential conservation hotspots. We then used regression analyses and Wilcoxon signed-rank tests to examine species-specific basin-wide and local abundance trends and identify species of potential conservation concern.
Results
Dispersion of sites in species abundance space was significantly greater in the 1900s compared with the 2000s, indicating homogenization had occurred. Of 36 native species analysed, 44.4% (16) showed basin-wide declines. Non-native species exhibited mixed patterns; site-level abundance increased in 2 of 15 species analysed (13%).
Main conclusions
Our results indicate basin-wide community homogenization has occurred within the UMNR, but many unique and diverse communities persist. If conserved, these could help maintain regional fish diversity. We found basin-wide declines in four endemic species, as well as spread patterns of non-native and native species that were not detected by a presence–absence analysis applied within the same study area. This finding illustrates the importance of considering both species’ abundance and occurrence patterns as separate dimensions of biodiversity to inform conservation planning.
Published version
application/pdf
en
Wiley
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
A new composite abundance metric detects stream fish declines and community homogenization during six decades of invasions
Diversity and Distributions
Article - Refereed
Text
ORIGINAL
ddi.13393.pdf
ddi.13393.pdf
Published version
application/pdf
2496782
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license.txt
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ddi.13393.pdf.txt
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MD5
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10919/104647
oai:vtechworks.lib.vt.edu:10919/104647
2023-11-29 19:07:02.99
http://creativecommons.org/licenses/by/4.0/
Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
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oai:vtechworks.lib.vt.edu:10919/820492020-11-25T15:06:13Zcom_10919_78629com_10919_78628com_10919_23829com_10919_5553col_10919_78791col_10919_23830
MacLean, Evan L.
Hare, Brian
Nunn, Charles L.
Addessi, Elsa
Amici, Federica
Anderson, Rindy C.
Aureli, Filippo
Baker, Joseph M.
Bania, Amanda E.
Barnard, Allison M.
Boogert, Neeltje J.
Brannon, Elizabeth M.
Bray, Emily E.
Bray, Joel
Brent, Lauren J. N.
Burkart, Judith M.
Call, Josep
Cantlon, Jessica F.
Cheke, Lucy G.
Clayton, Nicola S.
Delgado, Mikel M.
DiVincenti, Louis J.
Fujita, Kazuo
Herrmann, Esther
Hiramatsu, Chihiro
Jacobs, Lucia F.
Jordan, Kerry E.
Laude, Jennifer R.
Leimgruber, Kristin L.
Messer, Emily J. E.
Moura, Antonio C. de A.
Ostojic, Ljerka
Picard, Alejandra
Platt, Michael L.
Plotnik, Joshua M.
Range, Friederike
Reader, Simon M.
Reddy, Rachna B.
Sandel, Aaron A.
Santos, Laurie R.
Schumann, Katrin
Seed, Amanda M.
Sewall, Kendra B.
Shaw, Rachael C.
Slocombe, Katie E.
Su, Yanjie
Takimoto, Ayaka
Tan, Jingzhi
Tao, Ruoting
van Schaik, Carel P.
Viranyi, Zsoia
Visalberghi, Elisabetta
Wade, Jordan C.
Watanabe, Arii
Widness, Jane
Young, Julie K.
Zentall, Thomas R.
Zhao, Yini
Biological Sciences
2018-02-09T17:56:03Z
2018-02-09T17:56:03Z
2014-04-21
http://hdl.handle.net/10919/82049
https://doi.org/10.1073/pnas.1323533111
111
20
Cognition presents evolutionary research with one of its greatest challenges. Cognitive evolution has been explained at the proximate level by shifts in absolute and relative brain volume and at the ultimate level by differences in social and dietary complexity. However, no study has integrated the experimental and phylogenetic approach at the scale required to rigorously test these explanations. Instead, previous research has largely relied on various measures of brain size as proxies for cognitive abilities. We experimentally evaluated these major evolutionary explanations by quantitatively comparing the cognitive performance of 567 individuals representing 36 species on two problem-solving tasks measuring self-control. Phylogenetic analysis revealed that absolute brain volume best predicted performance across species and accounted for considerably more variance than brain volume controlling for body mass. This result corroborates recent advances in evolutionary neurobiology and illustrates the cognitive consequences of cortical reorganization through increases in brain volume. Within primates, dietary breadth but not social group size was a strong predictor of species differences in self-control. Our results implicate robust evolutionary relationships between dietary breadth, absolute brain volume, and self-control. These findings provide a significant first step toward quantifying the primate cognitive phenome and explaining the process of cognitive evolution.
We thank Natalie Cooper and Sunil Suchindran for statistical advice; Jeff Stevens and two anonymous reviewers for comments on drafts of this manuscript; and Ikuma Adachi, Nathan Emery, Daniel Haun, Marc Hauser, Ludwig Huber, Al Kamil, Chris Krupenye, Luke Matthews, Collin McCabe, Alexandra Rosati, Kara Schroepfer, Jeff Stevens, Tara Stoinski, Michael Tomasello, and Victoria Wobber for their helpful discussion during the workshops from which this research emerged. F. Aureli and F. Amici thank Iber Rodriguez Castillo, Roberto Pacheco Mendez, Fernando Victoria Arceo, Liesbeth Sterck, Barbara Tiddi, and all the animal keepers at the facilities where the data were collected for support and cooperation. K.E.S. and A.P. thank Steve Nichols and the staff at The Parrot Zoo. This work was supported by the National Evolutionary Synthesis Center (NESCent) through support of a working group led by C.L.N. and B.H. NESCent is supported by the National Science Foundation (NSF) EF-0905606. For training in phylogenetic comparative methods, we thank the AnthroTree Workshop (supported by NSF BCS-0923791). Y.S. thanks the National Natural Science Foundation of China (Project 31170995) and National Basic Research Program (973 Program: 2010CB833904). E.E.B. thanks the Duke Vertical Integration Program and the Duke Undergraduate Research Support Office. J.M.P. was supported by a Newton International Fellowship from the Royal Society and the British Academy. L.R.S. thanks the James S. McDonnell Foundation for Award 220020242. L.J.N.B. and M.L.P. acknowledge the National Institutes of Mental Health (R01-MH096875 and R01-MH089484), a Duke Institute for Brain Sciences Incubator Award (to M.L.P.), and a Duke Center for Interdisciplinary Decision Sciences Fellowship (to L.J.N.B.). E.V. and E.A. thank the Programma Nazionale per la Ricerca–Consiglio Nazionale delle Ricerche (CNR) Aging Program 2012–2014 for financial support, Roma Capitale– Museo Civico di Zoologia and Fondazione Bioparco for hosting the Istituto di Scienze e Tecnologie della Cognizione–CNR Unit of Cognitive Primatology and Primate Centre, and Massimiliano Bianchi and Simone Catarinacci for assistance with capuchin monkeys. K.F. thanks the Japan Society for the Promotion of Science (JSPS) for Grant-in-Aid for Scientific Research 20220004. F. Aureli thanks the Stages in the Evolution and Development of Sign Use project (Contract 012-984 NESTPathfinder) and the Integrating Cooperation Research Across Europe project (Contract 043318), both funded by the European Community’s Sixth Framework Programme (FP6/2002– 2006). F. Amici was supported by Humboldt Research Fellowship for Postdoctoral Researchers (Humboldt ID 1138999). L.F.J. and M.M.D. acknowledge NSF Electrical, Communications, and Cyber Systems Grant 1028319 (to L.F.J.) and an NSF Graduate Fellowship (to M.M.D.). C.H. thanks Grant-in-Aid for JSPS Fellows (10J04395). A.T. thanks Research Fellowships of the JSPS for Young Scientists (21264). F.R. and Z.V. acknowledge Austrian Science Fund (FWF) Project P21244-B17, the European Research Council (ERC) under the European Union’s Seventh Framework Programme (FP/2007–2013)/ERC Grant Agreement 311870 (to F.R.), Vienna Science and Technology Fund Project CS11-026 (to Z.V.), and many private sponsors, including Royal Canin for financial support and the Game Park Ernstbrunn for hosting the Wolf Science Center. S.M.R. thanks the Natural Sciences and Engineering Research Council (Canada). J.K.Y. thanks the US Department of Agriculture–Wildlife Services– National Wildlife Research Center. J.F.C. thanks the James S. McDonnell Foundation and Alfred P. Sloan Foundation. E.L.M. and B.H. thank the Duke Lemur Center and acknowledge National Institutes of Health Grant 5 R03 HD070649-02 and NSF Grants DGE-1106401, NSF-BCS-27552, and NSF-BCS- 25172. This is Publication 1265 of the Duke Lemur Center.
en_US
NAS
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
psychology
behavior
comparative methods
inhibitory control
executive function
The evolution of self-control
PNAS
Article - Refereed
ORIGINAL
SewallSelf-Control2014.pdf
SewallSelf-Control2014.pdf
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MD5
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LICENSE
license.txt
license.txt
text/plain; charset=utf-8
1535
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TEXT
SewallSelf-Control2014.pdf.txt
SewallSelf-Control2014.pdf.txt
Extracted text
text/plain
75396
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10919/82049
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/82049
2020-11-25 15:06:13.553
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/797942020-10-22T03:25:28Zcom_10919_78629com_10919_78628com_10919_23829com_10919_5553col_10919_78630col_10919_23830
Hipsey, Matthew R.
Hamilton, David P.
Hanson, Paul C.
Carey, Cayelan C.
Coletti, Janaine Z.
Read, Jordan S.
Ibelings, Bas W.
Valesini, Fiona J.
Brookes, Justin D.
Biological Sciences
2017-10-26T13:33:05Z
2017-10-26T13:33:05Z
2015-09-02
http://hdl.handle.net/10919/79794
https://doi.org/10.1002/2015WR017175
51
Maintaining the health of aquatic systems is an essential component of sustainable catchment management, however, degradation of water quality and aquatic habitat continues to challenge scientists and policy-makers. To support management and restoration efforts aquatic system models are required that are able to capture the often complex trajectories that these systems display in response to multiple stressors. This paper explores the abilities and limitations of current model approaches in meeting this challenge, and outlines a strategy based on integration of flexible model libraries and data from observation networks, within a learning framework, as a means to improve the accuracy and scope of model predictions. The framework is comprised of a data assimilation component that utilizes diverse data streams from sensor networks, and a second component whereby model structural evolution can occur once the model is assessed against theoretically relevant metrics of system function. Given the scale and transdisciplinary nature of the prediction challenge, network science initiatives are identified as a means to develop and integrate diverse model libraries and workflows, and to obtain consensus on diagnostic approaches to model assessment that can guide model adaptation. We outline how such a framework can help us explore the theory of how aquatic systems respond to change by bridging bottom-up and top-down lines of enquiry, and, in doing so, also advance the role of prediction in aquatic ecosystem management.
This work was funded by the Australian Research Council grant DP130104078.
en_US
American Geophysical Union
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Predicting the resilience and recovery of aquatic systems: A framework for model evolution within environmental observatories
Water Resources Research
Article - Refereed
ORIGINAL
Hipsey_et_al-2015-Water_Resources_Research.pdf
Hipsey_et_al-2015-Water_Resources_Research.pdf
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license.txt
license.txt
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MD5
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TEXT
Hipsey_et_al-2015-Water_Resources_Research.pdf.txt
Hipsey_et_al-2015-Water_Resources_Research.pdf.txt
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text/plain
104246
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THUMBNAIL
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MD5
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10919/79794
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/79794
2020-10-22 03:25:28.063
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/736262024-03-13T14:09:53Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553col_10919_70873col_10919_23830
Battogtokh, D.
Tyson, John J.
Biological Sciences
2016-12-09T21:32:17Z
2016-12-09T21:32:17Z
2004-09-01
Battogtokh, D.; Tyson, J. J., "bifurcation analysis of a model of the budding yeast cell cycle," Chaos 14, 653 (2004); http://dx.doi.org/10.1063/1.1780011
1054-1500
http://hdl.handle.net/10919/73626
https://doi.org/10.1063/1.1780011
14
3
We study the bifurcations of a set of nine nonlinear ordinary differential equations that describe regulation of the cyclin-dependent kinase that triggers DNA synthesis and mitosis in the budding yeast, Saccharomyces cerevisiae. We show that Clb2-dependent kinase exhibits bistability (stable steady states of high or low kinase activity). The transition from low to high Clb2-dependent kinase activity is driven by transient activation of Cln2-dependent kinase, and the reverse transition is driven by transient activation of the Clb2 degradation machinery. We show that a four-variable model retains the main features of the nine-variable model. In a three-variable model exhibiting birhythmicity (two stable oscillatory states), we explore possible effects of extrinsic fluctuations on cell cycle progression. (C) 2004 American Institute of Physics.
DARPA's Biocomputation Program (AFRL #F300602-02-0572)
Published version
653 - 661 (9) page(s)
English
American Institute of Physics
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000223904400015&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Mathematics, Applied
Physics, Mathematical
Mathematics
Physics
MATHEMATICS, APPLIED
PHYSICS, MATHEMATICAL
MATHEMATICAL-MODEL
MOLECULAR-MODEL
FISSION YEAST
DYNAMICS
NOISE
BIRHYTHMICITY
OSCILLATIONS
TRANSITIONS
HYSTERESIS
CIRCUITS
Bifurcation analysis of a model of the budding yeast cell cycle
Chaos
Article - Refereed
/Virginia Tech
/Virginia Tech/All T&R Faculty
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Science/COS T&R Faculty
/Virginia Tech/University Distinguished Professors
ORIGINAL
0404006v1.pdf
Draft Version
application/pdf
604113
https://vtechworks.lib.vt.edu/bitstreams/cc2422d5-9a1d-41f3-b6b7-5c7595525866/download
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MD5
1
LICENSE
VTUL_Distribution_License_2016_05_09.pdf
application/pdf
18524
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MD5
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0404006v1.pdf.txt
0404006v1.pdf.txt
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0404006v1.pdf.jpg
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oai:vtechworks.lib.vt.edu:10919/73626
2024-03-13 14:09:53.815
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
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oai:vtechworks.lib.vt.edu:10919/250932022-03-29T18:37:35Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:32Z
2014-01-23T02:36:32Z
2011
http://hdl.handle.net/10919/25093
http://www.johncairns.net/Commentaries/eliminating.pdf
Scientific evidence on climate change, including global warming, is more abundant than ever, but politicians and citizens are rejecting or attacking scientists for the evidence they have found. Science is reality based on a preponderance of evidence, and reality is often hard to accept. The biggest threat to human security is not terrorists or nuclear weapons, but the results of what would happen if humankind continues to ignore the warning signs of the biosphere.
en_US
Virginia Tech
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Cairns Jr., John
climate change
Sustainability
uncertainty
Why is Humankind Eliminating its Future
Article
Text
ORIGINAL
cairns_humankind_eliminating_future.pdf
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In Copyright
open.access
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VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1145012023-04-14T07:12:51Zcom_10919_23829com_10919_5553col_10919_23830
Rebholz, Zarley
Lancaster, Jason
Larose, Hailey
Khrimian, Ashot
Luck, Katrin
Sparks, Michael E.
Gendreau, Kerry L.
Shewade, Leena
Koellner, Tobias G.
Weber, Donald C.
Gundersen-Rindal, Dawn E.
O'Maille, Paul
Morozov, Alexandre, V
Tholl, Dorothea
2023-04-13T19:35:09Z
2023-04-13T19:35:09Z
2023-01
103879
http://hdl.handle.net/10919/114501
https://doi.org/10.1016/j.ibmb.2022.103879
152
36470318
1879-0240
Insects use diverse arrays of small molecules such as metabolites of the large class of terpenes for intra-and inter -specific communication and defense. These molecules are synthesized by specialized metabolic pathways; however, the origin of enzymes involved in terpene biosynthesis and their evolution in insect genomes is still poorly understood. We addressed this question by investigating the evolution of isoprenyl diphosphate synthase (IDS)-like genes with terpene synthase (TPS) function in the family of stink bugs (Pentatomidae) within the large order of piercing-sucking Hemipteran insects. Stink bugs include species of global pest status, many of which emit structurally related 15-carbon sesquiterpenes as sex or aggregation pheromones. We provide evidence for the emergence of IDS-type TPS enzymes at the onset of pentatomid evolution over 100 million years ago, coinciding with the evolution of flowering plants. Stink bugs of different geographical origin maintain small IDS -type families with genes of conserved TPS function, which stands in contrast to the diversification of TPS genes in plants. Expanded gene mining and phylogenetic analysis in other hemipteran insects further provides evidence for an ancient emergence of IDS-like genes under presumed selection for terpene-mediated chemical interactions, and this process occurred independently from a similar evolution of IDS-type TPS genes in beetles. Our findings further suggest differences in TPS diversification in insects and plants in conjunction with different modes of gene functionalization in chemical interactions.
National Science Foundation; US Department of Agriculture National Institute of Food and Agriculture; [MCB1920914]; [MCB1920922]; [MCB1920925]; [2016-67013-24759]
Published version
Acknowledgements This work was supported by the National Science Foundation (MCB1920914, MCB1920922, MCB1920925) to AVM, P.O., and D.T., respectively, and the US Department of Agriculture National Institute of Food and Agriculture (2016-67013-24759) to DT, AK, DEG-R, and DCW. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. The USDA is an equal opportunity provider and employer.
application/pdf
en
Pergamon-Elsevier Science
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
Pheromone
Terpene
Terpene synthase
Isoprenyl diphosphate synthase
Pentatomid
Hemiptera
Ancient origin and conserved gene function in terpene pheromone and defense evolution of stink bugs and hemipteran insects
Insect Biochemistry and Molecular Biology
Article - Refereed
Text
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1-s2.0-S0965174822001618-main.pdf
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oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/114501
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Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
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vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/736342023-06-21T19:15:34Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553com_10919_18738com_10919_5539com_10919_23261col_10919_70873col_10919_23830col_10919_23145col_10919_23262
Barik, Debashis
Baumann, William T.
Paul, Mark R.
Novak, Bela
Tyson, John J.
Electrical and Computer Engineering
Mechanical Engineering
Biological Sciences
2016-12-09T21:35:55Z
2016-12-09T21:35:55Z
2010-08-01
1744-4292
http://hdl.handle.net/10919/73634
https://doi.org/10.1038/msb.2010.55
6
In order for the cell’s genome to be passed intact from one generation to the next, the events of the cell cycle (DNA replication, mitosis, cell division) must be executed in the correct order, despite the considerable molecular noise inherent in any protein-based regulatory system residing in the small confines of a eukaryotic cell. To assess the effects of molecular fluctuations on cell-cycle progression in budding yeast cells, we have constructed a new model of the regulation of Cln- and Clb-dependent kinases, based on multisite phosphorylation of their target proteins and on positive and negative feedback loops involving the kinases themselves. To account for the significant role of noise in the transcription and translation steps of gene expression, the model includes mRNAs as well as proteins. The model equations are simulated deterministically and stochastically to reveal the bistable switching behavior on which proper cell-cycle progression depends and to show that this behavior is robust to the level of molecular noise expected in yeast-sized cells (B50 fL volume). The model gives a quantitatively accurate account of the variability observed in the G1-S transition in budding yeast, which is governed by an underlying sizer + timer control system.
Published version
? - ? (18) page(s)
application/pdf
en
Nature Publishing Group
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000284527700002&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported
http://creativecommons.org/licenses/by-nc-sa/3.0/
Biochemistry & Molecular Biology
bistability
cell-cycle variability
size control
stochastic model
transcription-translation coupling
ANAPHASE-PROMOTING COMPLEX
SACCHAROMYCES-CEREVISIAE
BUDDING YEAST
G1 CYCLINS
PHOSPHATASE CDC14
GENE-EXPRESSION
S-PHASE
G1-SPECIFIC TRANSCRIPTION
PROTEIN-PHOSPHORYLATION
POSITIVE FEEDBACK
A model of yeast cell-cycle regulation based on multisite phosphorylation
Molecular Systems Biology
Article - Refereed
Text
/Virginia Tech
/Virginia Tech/All T&R Faculty
/Virginia Tech/Engineering
/Virginia Tech/Engineering/COE T&R Faculty
/Virginia Tech/Engineering/Electrical and Computer Engineering
/Virginia Tech/Engineering/Mechanical Engineering
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Science/COS T&R Faculty
/Virginia Tech/University Distinguished Professors
ORIGINAL
A model of yeast cell-cycle regulation based on multisite phosphorylation.pdf
Publisher's Version
application/pdf
2371050
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/568c6eca-e8fb-4327-838d-c3213909e082/download
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MD5
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VTUL_Distribution_License_2016_05_09.pdf
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MD5
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TEXT
A model of yeast cell-cycle regulation based on multisite phosphorylation.pdf.txt
A model of yeast cell-cycle regulation based on multisite phosphorylation.pdf.txt
Extracted text
text/plain
84409
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b474ac4a4ce5f1b2b3e736c82a09fbc8
MD5
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THUMBNAIL
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MD5
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10919/73634
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/73634
2023-06-21 19:15:34.332
http://creativecommons.org/licenses/by-nc-sa/3.0/
Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1132332023-11-29T11:24:08Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553com_10919_25796col_10919_70873col_10919_23830col_10919_25797
Mosig, Rebecca A.
Kojima, Shihoko
2023-01-18T14:28:08Z
2023-01-18T14:28:08Z
2022-06-01
2023-01-17T16:50:53Z
1084-9521
S1084-9521(21)00093-8 (PII)
http://hdl.handle.net/10919/113233
https://doi.org/10.1016/j.semcdb.2021.04.020
126
Kojima, Shihoko [0000-0001-9579-7830]
34116930
1096-3634
Long non-coding RNAs (lncRNAs) are a new class of regulatory RNAs that play important roles in disease development and a variety of biological processes. Recent studies have underscored the importance of lncRNAs in the circadian clock system and demonstrated that lncRNAs regulate core clock genes and the core clock machinery in mammals. In this review, we provide an overview of our current understanding of how lncRNAs regulate the circadian clock without coding a protein. We also offer additional insights into the challenges in understanding the functions of lncRNAs and other unresolved questions in the field. We do not cover other regulatory ncRNAs even though they also play important roles; readers are highly encouraged to refer to other excellent reviews on this topic.
Accepted version
Pages 79-86
8 page(s)
application/pdf
en
Academic Press-Elsevier
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000799035500010&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Long non-coding RNA
Antisense transcript
Circadian rhythms
Molecular clock
PINEAL CLOCK EXPRESSION
GENE-EXPRESSION
TRANSCRIPTIONAL ARCHITECTURE
ANTISENSE RNA
EVOLUTION
REVEALS
ROLES
INDUCTION
LANDSCAPE
MPER1
Neurosciences
Biotechnology
Genetics
Sleep Research
Generic health relevance
Animals
Mammals
RNA, Untranslated
Circadian Rhythm
Circadian Clocks
RNA, Long Noncoding
Timing without coding: How do long non-coding RNAs regulate circadian rhythms?
Seminars in Cell & Developmental Biology
Article - Refereed
Journal
Text
2021-04-21
/Virginia Tech
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/All T&R Faculty
/Virginia Tech/Science/COS T&R Faculty
SWORD
sword-2023-01-17T11:50:53.original.xml
Original SWORD entry document
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130
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ORIGINAL
2021-4-12_Manuscript_UnMarked.pdf
2021-4-12_Manuscript_UnMarked.pdf
Accepted version
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2021-4-12_Manuscript_UnMarked.pdf.txt
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MD5
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10919/113233
oai:vtechworks.lib.vt.edu:10919/113233
2023-11-29 11:24:08.947
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1001622021-06-21T19:04:57Zcom_10919_8195com_10919_25799com_10919_23829com_10919_5553com_10919_19035com_10919_5539col_10919_18629col_10919_23830col_10919_24290
Xu, Chunrui
Weston, Bronson R.
Tyson, John J.
Cao, Yang
Biological Sciences
Computer Science
2020-10-05T12:34:25Z
2020-10-05T12:34:25Z
2020-09-30
2020-10-04T03:28:54Z
BMC Bioinformatics. 2020 Sep 30;21(Suppl 14):408
http://hdl.handle.net/10919/100162
https://doi.org/10.1186/s12859-020-03687-z
Background
Second messengers, c-di-GMP and (p)ppGpp, are vital regulatory molecules in bacteria, influencing cellular processes such as biofilm formation, transcription, virulence, quorum sensing, and proliferation. While c-di-GMP and (p)ppGpp are both synthesized from GTP molecules, they play antagonistic roles in regulating the cell cycle. In C. crescentus, c-di-GMP works as a major regulator of pole morphogenesis and cell development. It inhibits cell motility and promotes S-phase entry by inhibiting the activity of the master regulator, CtrA. Intracellular (p)ppGpp accumulates under starvation, which helps bacteria to survive under stressful conditions through regulating nucleotide levels and halting proliferation. (p)ppGpp responds to nitrogen levels through RelA-SpoT homolog enzymes, detecting glutamine concentration using a nitrogen phosphotransferase system (PTS Ntr). This work relates the guanine nucleotide-based second messenger regulatory network with the bacterial PTS Ntr system and investigates how bacteria respond to nutrient availability.
Results
We propose a mathematical model for the dynamics of c-di-GMP and (p)ppGpp in C. crescentus and analyze how the guanine nucleotide-based second messenger system responds to certain environmental changes communicated through the PTS Ntr system. Our mathematical model consists of seven ODEs describing the dynamics of nucleotides and PTS Ntr enzymes. Our simulations are consistent with experimental observations and suggest, among other predictions, that SpoT can effectively decrease c-di-GMP levels in response to nitrogen starvation just as well as it increases (p)ppGpp levels. Thus, the activity of SpoT (or its homologues in other bacterial species) can likely influence the cell cycle by influencing both c-di-GMP and (p)ppGpp.
Conclusions
In this work, we integrate current knowledge and experimental observations from the literature to formulate a novel mathematical model. We analyze the model and demonstrate how the PTS Ntr system influences (p)ppGpp, c-di-GMP, GMP and GTP concentrations. While this model does not consider all aspects of PTS Ntr signaling, such as cross-talk with the carbon PTS system, here we present our first effort to develop a model of nutrient signaling in C. crescentus.
Published version
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
The Author(s)
Cell cycle control and environmental response by second messengers in Caulobacter crescentus
BMC Bioinformatics
Article - Refereed
Text
StillImage
ORIGINAL
12859_2020_Article_3687.pdf
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2289469
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MD5
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LICENSE
license.txt
license.txt
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0
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MD5
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SWORD
art_1751977641900806950.zip
SWORD deposit package
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MD5
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TEXT
12859_2020_Article_3687.pdf.txt
12859_2020_Article_3687.pdf.txt
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58537
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MD5
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10919/100162
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/100162
2021-06-21 19:04:57.812
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1026472021-03-24T14:08:47Zcom_10919_23829com_10919_5553col_10919_23830
Kaiser, April L.
Soule, Peter
van de Gevel, Saskia L.
Knapp, Paul
Bhuta, Arvind
Walters, Jeffrey R.
Montpellier, Evan
Biological Sciences
2021-03-10T13:52:43Z
2021-03-10T13:52:43Z
2020-10-01
0378-1127
118291
http://hdl.handle.net/10919/102647
https://doi.org/10.1016/j.foreco.2020.118291
473
1872-7042
Old-growth longleaf pine (Pines palustris) is a keystone/foundation species for 29 threatened or endangered species in the Coastal Plain of the southeastern United States. The endangered red-cockaded woodpecker (Dryobates borealis; RCW) and endangered longleaf pine have an established ecological association. Here, we explore differences in climate/growth response and radial growth disturbance events in trees with RCW cavities compared to non-cavity trees in the Sandhills Gameland Reserve in North Carolina, USA. Using standard dendrochronological techniques, we collected and analyzed core samples from trees selected by RCW for their cavities (RCWC) and adjacent control trees (RCWCo) that had no visible cavity. We developed RCWC and RCWCo tree-ring chronologies that allowed us to examine if climate vulnerability is a component of the RCW selection process for their nests. Specifically, we investigated climate/growth responses, radial growth suppressions, and physical characteristics of both tree types through a comparison of tree age, latewood radial growth measurements, and number of resin ducts. For long-term climate response (1910-2018), we found no significant differences between RCWC and RCWCo trees. However, we identified temporal differences in climate/growth relationships between RCWC and RCWCo as well as significant differences in the number of suppression events and spatially-grouped suppression events. For tree physiology, we found more resin ducts during 1950-2018 in RCWC trees. Our dendroecological-based investigation examines multiple factors in addressing the question of why RCWs select specific longleaf pine trees for cavities, which may help improve conservation efforts for RCW and longleaf pine.
Appalachian State University
We thank Brady Beck from the North Carolina Wildlife Resources Commission's Sandhills Gameland. Additionally, we thank Tyler Mitchell for help with field work and Julia Adams, Brianna Hibner, and Rebecca Sigafoos for help with processing the collected cores. We also extend great gratitude to Dr. Jan Altman at the Czech Academy of Sciences for crucial R code that made this study possible. We also thank Dr. Chris Guiterman for R code that helped us illustrate our suppression data. The authors received funding for this project from Appalachian State University.
Public domain – authored by a U.S. government employee
application/pdf
en
Public Domain
http://creativecommons.org/publicdomain/mark/1.0/
Red-cockaded woodpecker
Longleaf pine
Dendroecology
Climate/growth sensitivity
Ecological disturbance
Dendroecological investigation of red-cockaded woodpecker cavity tree selection in endangered longleaf pine forests
Forest Ecology and Management
Article - Refereed
Text
StillImage
ORIGINAL
1-s2.0-S0378112720310604-main.pdf
1-s2.0-S0378112720310604-main.pdf
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3367381
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MD5
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TEXT
1-s2.0-S0378112720310604-main.pdf.txt
1-s2.0-S0378112720310604-main.pdf.txt
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text/plain
74681
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MD5
2
THUMBNAIL
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1-s2.0-S0378112720310604-main.pdf.jpg
IM Thumbnail
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https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/01431b20-166c-4cbf-ba68-5e9ec73ce885/download
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MD5
3
10919/102647
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/102647
2021-03-24 14:08:47.967
http://creativecommons.org/publicdomain/mark/1.0/
Public Domain
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/250832020-11-12T20:37:02Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:31Z
2014-01-23T02:36:31Z
2009
http://hdl.handle.net/10919/25083
http://www.johncairns.net/Papers/Is%20Homo%20sapiens%20Just%20Another%20Stochastic.pdf
Surprisingly, mass extinctions probably account for the disappearance of less than 5% of all extinct species _ 95% of species extinctions occur between mass extinctions. In short, extinction can occur at any time in Earth s history. The concept of sustainable use of the planet assumes that humans can live on the planet indefinitely _ or at least until the sun dies. However, human production of greenhouse gases is resulting in rapid climate change that threatens human society globally. In addition, greenhouse gas emissions are increasing markedly, and no agreement has been reached on specific goals for reducing emissions. Finally, coal is being used to replace diminishing supplies of petroleum, despite evidence that coal produces about twice as many greenhouse gas emissions per unit of energy produced than petroleum. Although alternative sources of energy (e.g., wind, solar, geothermal) are becoming increasingly popular, humankind s primary source of energy is fossil fuels. Global climate change resulting from anthropogenic greenhouse gas emissions has already affected food production, water supplies, natural habitat, and human security. Climate change and other types of habitat destruction threaten the biospheric life support system upon which human survival depends. All these issues suggest that Homo sapiens may be just another stochastic event that is adversely affecting life on Earth.
en_US
Asian Journal of Experimental Sciences
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
extinction of Homo sapiens
climate instability
major global extinctions
social evolution
ani-intellectualism
short time for social change
Is Homo sapiens Just Another Stochastic Event in the History of Life on Earth
Article
Text
ORIGINAL
cairns_homo_sapiens.pdf
application/pdf
53912
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MD5
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TEXT
cairns_homo_sapiens.pdf.txt
cairns_homo_sapiens.pdf.txt
Extracted text
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27087
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MD5
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10919/25083
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/25083
2020-11-12 20:37:02.875
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/253432020-10-28T05:42:01Zcom_10919_23829com_10919_5553col_10919_23830
Virginia Tech
Opell, B. D.
Schwend, H. S.
Vito, S. T.
Biological Sciences
2014-02-07T18:29:40Z
2014-02-07T18:29:40Z
2011-07
2014-02-04
Opell, Brent D.; Schwend, Harold S.; Vito, Stephen T., "Constraints on the adhesion of viscous threads spun by orb-weaving spiders: the tensile strength of glycoprotein glue exceeds its adhesion," J Exp Biol 214, 2237-2241 (2011); doi: 10.1242/_jeb.055707
0022-0949
http://hdl.handle.net/10919/25343
http://jeb.biologists.org/content/214/13/2237.full.pdf+html
https://doi.org/10.1242/jeb.055707
In this study we tested the hypothesis that a viscous thread releases its hold on a surface because its glycoprotein glue pulls from the surface and not because its elongating droplets break near their attachment to the surface. We compared the values obtained when three species' viscous threads adhered to four smooth surfaces, which differed in their total surface energy and in the proportions of their dispersion and polar energy components. Although water comprised 43-70% of the volume of these viscous droplets, only the dispersion surface energies of test materials and not their polar surface energies impacted thread adhesion. These results support the droplet pull-off hypothesis and are consistent with a previous finding that capillary force contributes little to thread adhesion. Just as a viscous thread's stickiness is constrained by the tensile strength of its supporting axial fibers, our findings suggest that glycoprotein adhesion is constrained by glycoprotein tensile strength.
National Science Foundation IOB-0445137
en_US
Company of Biologists Ltd.
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
The Company of Biologists Ltd
Araneioidea
dispersion surface energy
Leucauge venusta
Metepeira
labyrinthea
Micrathena gracilis
polar surface energy
CAPTURE THREADS
CORONA DISCHARGE
SURFACE-ENERGY
WEB
WATER
Constraints on the adhesion of viscous threads spun by orb-weaving spiders: the tensile strength of glycoprotein glue exceeds its adhesion
Journal of Experimental Biology
Article - Refereed
ORIGINAL
J Exp Biol-2011-Opell-2237-41.pdf
J Exp Biol-2011-Opell-2237-41.pdf
Main article
application/pdf
677141
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MD5
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TEXT
J Exp Biol-2011-Opell-2237-41.pdf.txt
J Exp Biol-2011-Opell-2237-41.pdf.txt
Extracted text
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27540
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IM Thumbnail
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10919/25343
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/25343
2020-10-28 05:42:01.377
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/257822020-11-24T17:44:07Zcom_10919_23829com_10919_5553col_10919_23830
Virginia Tech
Csikasz-Nagy, A.
Battogtokh, D.
Chen, Katherine C.
Novak, Bela
Tyson, John J.
Biological Sciences
2014-02-26T19:10:06Z
2014-02-26T19:10:06Z
2006-06
2014-02-05
Csikasz-Nagy, A; Battogtokh, D; Chen, KC; et al. "Analysis of a generic model of eukaryotic cell-cycle regulation," Biophysical Journal 90(12), 4361-4379 (2006); doi: 10.1529/biophysj.106.081240
0006-3495
http://hdl.handle.net/10919/25782
http://www.sciencedirect.com/science/article/pii/S0006349506726145
https://doi.org/10.1529/biophysj.106.081240
We propose a protein interaction network for the regulation of DNA synthesis and mitosis that emphasizes the universality of the regulatory system among eukaryotic cells. The idiosyncrasies of cell cycle regulation in particular organisms can be attributed, we claim, to specific settings of rate constants in the dynamic network of chemical reactions. The values of these rate constants are determined ultimately by the genetic makeup of an organism. To support these claims, we convert the reaction mechanism into a set of governing kinetic equations and provide parameter values (specific to budding yeast, fission yeast, frog eggs, and mammalian cells) that account for many curious features of cell cycle regulation in these organisms. Using one-parameter bifurcation diagrams, we show how overall cell growth drives progression through the cell cycle, how cell-size homeostasis can be achieved by two different strategies, and how mutations remodel bifurcation diagrams and create unusual cell-division phenotypes. The relation between gene dosage and phenotype can be summarized compactly in two-parameter bifurcation diagrams. Our approach provides a theoretical framework in which to understand both the universality and particularity of cell cycle regulation, and to construct, in modular fashion, increasingly complex models of the networks controlling cell growth and division.
Defense Advanced Research Project Agency AFRL F30602-02-0572
James S. McDonnell Foundation 21002050
European Commission COMBIO, LSHG-CT-503568
en_US
CELL PRESS
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Biophysical Society
protein-protein interactions
anaphase-promoting complex
apc-dependent
proteolysis
xenopus-oocyte extracts
m-phase control
fission yeast
saccharomyces-cerevisiae
mathematical-model
bifurcation-analysis
dna-replication
Analysis of a generic model of eukaryotic cell-cycle regulation
Biophysical Journal
Article - Refereed
TEXT
1-s2.0-S0006349506726145-main.pdf.txt
1-s2.0-S0006349506726145-main.pdf.txt
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MD5
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IM Thumbnail
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MD5
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ORIGINAL
1-s2.0-S0006349506726145-main.pdf
1-s2.0-S0006349506726145-main.pdf
Main article
application/pdf
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MD5
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10919/25782
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/25782
2020-11-24 17:44:07.003
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/852242023-04-14T17:49:47Zcom_10919_23829com_10919_5553col_10919_23830
Wojdak, Jeremy M.
Edman, Robert M.
Wyderko, Jennie A.
Zemmer, Sally A.
Belden, Lisa K.
Biological Sciences
2018-10-04T14:11:21Z
2018-10-04T14:11:21Z
2014-08-13
e105059
http://hdl.handle.net/10919/85224
https://doi.org/10.1371/journal.pone.0105059
9
8
25119568
1932-6203
Variation in host species composition can dramatically alter parasite transmission in natural communities. Whether diverse host communities dilute or amplify parasite transmission is thought to depend critically on species traits, particularly on how hosts affect each other’s densities, and their relative competency as hosts. Here we studied a community of potential hosts and/or decoys (i.e. non-competent hosts) for two trematode parasite species, Echinostoma trivolvis and Ribeiroia ondatrae, which commonly infect wildlife across North America. We manipulated the density of a focal host (green frog tadpoles, Rana clamitans), in concert with manipulating the diversity of alternative species, to simulate communities where alternative species either (1) replace the focal host species so that the total number of individuals remains constant (substitution) or (2) add to total host density (addition). For E. trivolvis, we found that total parasite transmission remained roughly equal (or perhaps decreased slightly) when alternative species replaced focal host individuals, but parasite transmission was higher when alternative species were added to a community without replacing focal host individuals. Given the alternative species were roughly equal in competency, these results are consistent with current theory. Remarkably, both total tadpole and per-capita tadpole infection intensity by E. trivolvis increased with increasing intraspecific host density. For R. ondatrae, alternative species did not function as effective decoys or hosts for parasite infective stages, and the diversity and density treatments did not produce clear changes in parasite transmission, although high tank to tank variation in R. ondatrae infection could have obscured patterns.
Published version
application/pdf
en
PLOS
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Host Density and Competency Determine the Effects of Host Diversity on Trematode Parasite Infection
PLOS ONE
Article - Refereed
Text
ORIGINAL
journal.pone.0105059.PDF
journal.pone.0105059.PDF
application/pdf
1834129
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MD5
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10919/85224
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/85224
2023-04-14 17:49:47.725
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/995792023-11-29T19:07:17Zcom_10919_5540com_10919_23829com_10919_5553com_10919_91436com_10919_5532col_10919_71752col_10919_23830col_10919_97229
Shrestha, Manisha
Bernhards, Robert C.
Fu, Yichen
Ryan, Kylie
Schubot, Florian D.
2020-08-06T17:58:43Z
2020-08-06T17:58:43Z
2020
2045-2322
http://hdl.handle.net/10919/99579
https://doi.org/10.1038/s41598-020-66555-z
10
1
32555263
The type III secretion system (T3SS) is a pivotal virulence mechanism of many Gram-negative bacteria. During infection, the syringe-like T3SS injects cytotoxic proteins directly into the eukaryotic host cell cytoplasm. In Pseudomonas aeruginosa, expression of the T3SS is regulated by a signaling cascade involving the proteins ExsA, ExsC, ExsD, and ExsE. The AraC-type transcription factor ExsA activates transcription of all T3SS-associated genes. Prior to host cell contact, ExsA is inhibited through direct binding of the anti-activator protein ExsD. Host cell contact triggers secretion of ExsE and sequestration of ExsD by ExsC to cause the release of ExsA. ExsA does not bind ExsD through the canonical ligand binding pocket of AraC-type proteins. Using site-directed mutagenesis and a specific in vitro transcription assay, we have now discovered that backbone interactions between the amino terminus of ExsD and the ExsA beta barrel constitute a pivotal part of the ExsD-ExsA interface. Follow-up bacterial two-hybrid experiments suggest additional contacts create an even larger protein– protein interface. The discovered role of the amino terminus of ExsD in ExsA binding explains how ExsC might relieve the ExsD-mediated inhibition of T3SS gene expression, because the same region of ExsD interacts with ExsC following host cell contact.
F.D.S.
8 pages
application/pdf
en
Springer Nature
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Backbone Interactions Between Transcriptional Activator ExsA and Anti-Activator ExsD Facilitate Regulation of the Type III Secretion System in Pseudomonas aeruginosa
Scientific Reports
2454-2156
400
Article - Refereed
Text
ORIGINAL
s41598-020-66555-z.pdf
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10919/99579
oai:vtechworks.lib.vt.edu:10919/99579
2023-11-29 19:07:17.46
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/995852023-11-29T19:07:19Zcom_10919_5540com_10919_23829com_10919_5553col_10919_71752col_10919_23830
Hope, Syndey F.
Kennamer, R. A.
Grimaudo, A. T.
Hallagan, John J.
Hopkins, William A.
Biological Sciences
2020-08-06T17:58:49Z
2020-08-06T17:58:49Z
2020
http://hdl.handle.net/10919/99585
https://doi.org/10.1093/iob/obaa003
Developmental conditions can have consequences for offspring fitness. For example, small changes (<1°C) in average avian incubation temperature have large effects on important post-hatch offspring phenotypes, including growth rate, thermoregulation, and behavior. Furthermore, average incubation temperatures differ among eggs within the same nest, to the extent (i.e., >1°C) that differences in offspring phenotypes within broods should result. A potential consequence of within-nest incubation temperature variation is inequality in behaviors that could cause differences in resource acquisition within broods. To investigate this, we incubated wood duck (Aix sponsa) eggs at one of two ecologically-relevant incubation temperatures (35°C or 36°C), formed mixed-incubation temperature broods after ducklings hatched, and conducted trials to measure duckling behaviors associated with acquisition of heat (one trial) or food (three trials). Contrary to our predictions, we found no effect of incubation temperature on duckling behaviors (e.g., time spent occupying heat source, frequency of feeding bouts). However, we found evidence that ducklings incubated at the higher temperature consumed more food during the 1-h feeding trials, and grew faster in body mass and structural size (culmen and tarsus) throughout the study, than those incubated at the lower temperature. Apparent food consumption during the trials was positively related to culmen length, suggesting that differences in food consumption may be driven by structural size. This could result in positive feedback, which would amplify size differences between offspring incubated at different temperatures. Thus, our study identifies incubation temperature as a mechanism by which fitness-related phenotypic differences can be generated and even amplified within avian broods.
This work was supported by the National Science Foundation Graduate Research Fellowship Program [478969]; the Virginia Tech Graduate Student Assembly Graduate Research Development Program; and the Institute for Critical Technology and Applied Science at Virginia Tech. This material is based upon work financially supported by the U.S. Department of Energy under Award Number DE-FC09-07SR22506 to the University of Georgia Research Foundation. The U.S. Department of Energy had no involvement in the study design, collection, analysis, and interpretation of the data, or in the decision to publish this article. Funding for Open Access was provided by Virginia Tech’s Open Access Subvention Fund.
18 pages
application/pdf
en
Oxford University Press
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Incubation Temperature Affects Duckling Body Size and Food Consumption Despite No Effect on Associated Feeding Behaviors
Integrative Organismal Biology
Article - Refereed
Text
ORIGINAL
obaa003.pdf
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25761728
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obaa003.pdf.txt
obaa003.pdf.txt
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84369
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10919/99585
oai:vtechworks.lib.vt.edu:10919/99585
2023-11-29 19:07:19.651
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/865422023-11-29T11:24:22Zcom_10919_23829com_10919_5553com_10919_25796col_10919_23830col_10919_25797
Kale, Shiv D.
Ayubi, Tariq
Chung, Dawoon
Tubau-Juni, Nuria
Leber, Andrew
Dang, Ha X.
Karyala, Saikumar
Hontecillas, Raquel
Lawrence, Christopher B.
Cramer, Robert A.
Bassaganya-Riera, Josep
Biological Sciences
Fralin Life Sciences Institute
2019-01-02T15:41:29Z
2019-01-02T15:41:29Z
2017-12-06
2045-2322
17096
http://hdl.handle.net/10919/86542
https://doi.org/10.1038/s41598-017-17000-1
7
Incidences of invasive pulmonary aspergillosis, an infection caused predominantly by Aspergillus fumigatus, have increased due to the growing number of immunocompromised individuals. While A. fumigatus is reliant upon deficiencies in the host to facilitate invasive disease, the distinct mechanisms that govern the host-pathogen interaction remain enigmatic, particularly in the context of distinct immune modulating therapies. To gain insights into these mechanisms, RNA-Seq technology was utilized to sequence RNA derived from lungs of 2 clinically relevant, but immunologically distinct murine models of IPA on days 2 and 3 post inoculation when infection is established and active disease present. Our findings identify notable differences in host gene expression between the chemotherapeutic and steroid models at the interface of immunity and metabolism. RT-qPCR verified model specific and nonspecific expression of 23 immune-associated genes. Deep sequencing facilitated identification of highly expressed fungal genes. We utilized sequence similarity and gene expression to categorize the A. fumigatus putative in vivo secretome. RT-qPCR suggests model specific gene expression for nine putative fungal secreted proteins. Our analysis identifies contrasting responses by the host and fungus from day 2 to 3 between the two models. These differences may help tailor the identification, development, and deployment of host-and/or fungal-targeted therapeutics.
Biocomplexity Institute of Virginia Tech; Office of the Director of National Intelligence (ODNI); Intelligence Advanced Research Projects Activity (IARPA), via the Army Research Office (ARO) [W911NF-17-2-0105]; HHS \ NIH \ National Institute of Allergy and Infectious Diseases (NIAID) [R01 AI081838]; Burroughs Wellcome Fund (BWF)
A portion of this study was funded through internal funding from the Biocomplexity Institute of Virginia Tech to SDK. This study is also based upon work supported by the Office of the Director of National Intelligence (ODNI), Intelligence Advanced Research Projects Activity (IARPA), via the Army Research Office (ARO) under cooperative Agreement Number W911NF-17-2-0105. The views and conclusions contained herein are those of the authors and should not be interpreted as necessarily representing the official policies or endorsements, either expressed or implied, of the ODNI, IARPA, ARO, or the U.S. Government. The efforts of Robert A Cramer were funded by HHS vertical bar NIH vertical bar National Institute of Allergy and Infectious Diseases (NIAID) (R01 AI081838). RAC holds an Investigators in the Pathogenesis of Infectious Diseases Award from the Burroughs Wellcome Fund (BWF).
25 pages
application/pdf
en_US
Springer Nature
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
c-type lectin
nitric-oxide synthase
beta-glucan receptor
fumigatus conidia
dendritic cells
in-vitro
hematopoietic transplantation
hematologic malignancies
corticosteroid treatment
cytokine production
Modulation of Immune Signaling and Metabolism Highlights Host and Fungal Transcriptional Responses in Mouse Models of Invasive Pulmonary Aspergillosis
Scientific Reports
Article - Refereed
Text
TEXT
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116683
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s41598-017-17000-1.pdf
s41598-017-17000-1.pdf
application/pdf
10543381
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MD5
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10919/86542
oai:vtechworks.lib.vt.edu:10919/86542
2023-11-29 11:24:22.738
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/766392023-11-29T19:07:20Zcom_10919_5540com_10919_23829com_10919_5553col_10919_71752col_10919_23830
Virginia Tech
Black, Wesley P.
Wang, Linglin
Davis, Manli Y.
Yang, Zhaomin
Biological Sciences
2017-03-13T17:13:41Z
2017-03-13T17:13:41Z
2015-12-07
http://hdl.handle.net/10919/76639
https://doi.org/10.1038/srep17831
5
Here we attempted to identify the downstream target of the DifE histidine kinase in the regulation of exopolysaccharide (EPS) production in the Gram-negative bacterium Myxococcus xanthus. This bacterium is an important model system for the studies of Type IV pilus (T4P) because it is motile by social (S) motility which is powered by T4P retraction. EPS is critical for S motility because it is the preferred anchor for T4P retraction in this bacterium. Previous studies identified the Dif chemosensory pathway as crucial for the regulation of EPS production. However, the downstream target of the DifE kinase in this pathway was unknown. In this study, EpsW, an orphan and single-domain response regulator (RR), was identified as a potential DifE target first by bioinformatics. Subsequent experiments demonstrated that epsW is essential for EPS biosynthesis in vivo and that EpsW is directly phosphorylated by DifE in vitro. Targted mutagenesis of epsW suggests that EpsW is unlikely the terminal RR of the Dif pathway. We propose instead that EpsW is an intermediary in a multistep phosphorelay that regulates EPS in M. xanthus.
application/pdf
en_US
Nature
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
The orphan response regulator EpsW is a substrate of the DifE kinase and it regulates exopolysaccharide in Myxococcus Xanthus
Scientific Reports
Article - Refereed
Text
ORIGINAL
ZhaominOrphan2015.pdf
ZhaominOrphan2015.pdf
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ZhaominOrphan2015.pdf.txt
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45485
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MD5
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10919/76639
oai:vtechworks.lib.vt.edu:10919/76639
2023-11-29 19:07:20.294
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1116982022-09-03T07:12:53Zcom_10919_23829com_10919_5553com_10919_19035com_10919_5539col_10919_23830col_10919_24290
Weston, Bronson R.
Tyson, John J.
Cao, Yang
2022-09-02T19:36:39Z
2022-09-02T19:36:39Z
2021-12-17
103413
http://hdl.handle.net/10919/111698
https://doi.org/10.1016/j.isci.2021.103413
24
12
34901785
2589-0042
In the alphaproteobacterium, Caulobacter crescentus, phosphorylated CtrA (CtrA similar to P), a master regulatory protein, binds directly to the chromosome origin (Cori) to inhibit DNA replication. Using a mathematical model of CtrA binding at Cori site [d], we provide computational evidence that CtrA(U) can disc ace CtrA similar to P from Cori at the G1-S transition. Investigation of this interaction within a detailed model of the C. crescentus cell cycle suggests that CckA phosphatase may clear Cori of CtrA similar to P by altering the [CtrA(U)]/[CtrA similar to P] ratio rather than by completely depleting CtrA similar to P. Model analysis reveals that the mechanism allows for a speedier transition into S phase, stabilizes the timing of chromosome replication under fluctuating rates of CtrA proteolysis, and may contribute to the viability of numerous mutant strains. Overall, these results suggest that CtrA(U) enhances the robustness of chromosome replication. More generally, our proposed regulation of CtrA:Cori dynamics may represent a novel motif for molecular signaling in cell physiology.
National Science Foundation (NSF) [CCF-1526666, MCB-1613741, CCF-1909122]
Published version
This work was partially supported by the National Science Foundation (NSF) under awards CCF-1526666, MCB-1613741, and CCF-1909122. The funding sources played no role in the design of the study, in the collection, analysis, and interpretation of data, and in writing the manuscript.
application/pdf
en
Cell Press
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
chromosome-replication origin
response regulator ctra
dna-replication
histidine kinase
polar localization
allosteric control
protease complex
temporal control
feedback-control
systems biology
Computational modeling of unphosphorylated CtrA:Cori binding in the Caulobacter cell cycle
iScience
Article - Refereed
Text
ORIGINAL
1-s2.0-S2589004221013845-main.pdf
1-s2.0-S2589004221013845-main.pdf
Published version
application/pdf
4726298
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MD5
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1-s2.0-S2589004221013845-main.pdf.txt
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10919/111698
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/111698
2022-09-03 07:12:53.813
http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/250872020-11-12T20:37:00Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:32Z
2014-01-23T02:36:32Z
2010
http://hdl.handle.net/10919/25087
http://www.johncairns.net/Papers/The%20Human%20Economy.pdf
For decades, deniers of the need to protect the environment have used the tactic of pitting the environment against the economy. However, the disingenuous tactic ignores the dependence of the human economy on the biosphere for raw materials (i.e., natural resources) and also that the present biospheric life support system has maintained conditions favorable (e.g., atmospheric gas balance) to the genus Homo for approximately 2 million years and for Homo sapiens for about 160,000 to 200,000 years. If this dependence were not significant, humans would become extinct and the human economy would disappear with them. Since the human economy is totally dependent upon the biosphere and humans are depenent on the biospheric life support system, why are they tolerant of the type of economic growth that damages the biosphere? The only reason that has some dubious validity is the assumption that, when a resource disappears, human ingenuity and creativity will be able to find a substitute for it. In short, deniers speculate that growth has no limits. However, in the latter part of the 20th century, ample evidence became available that this speculation was simply not true. In addition, persuasive evidence also substantiates that non-carbon alternative energy sources (e.g., solar, wind) would be less damaging to the environment and also be an economic stimulus. Humankind should only engage in activities that nurture the biosphere.
en_US
Asian Journal of Experimental Sciences
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
biosphere
life support system
economic growth
naturl resources
environment versus economy
environmental damage deniers
The Human Economy is a Subset of the Biosphere
Article
Text
ORIGINAL
cairns_human_economy.pdf
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35656
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MD5
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cairns_human_economy.pdf.txt
cairns_human_economy.pdf.txt
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8517
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MD5
4
10919/25087
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/25087
2020-11-12 20:37:00.902
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/249972020-11-12T20:37:02Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:25Z
2014-01-23T02:36:25Z
2005
http://hdl.handle.net/10919/24997
http://www.johncairns.net/Papers/sustemergency.pdf
Emergency situations require more resources than normal operations. If human society does not address environmental problems effectively before they reach the emergency stage, life from then on will be in a sustained (indefinite) state of emergency. Sustainable use of the planet requires optimal use of resources, which can be facilitated by avoiding sustained emergency conditions. Some emergencies beyond human control will always occur, and adequate global resources must be allocated to cope with them. However, a huge number of emergencies is simply the result of failing to take precautionary, preventative action in time.
en_US
International Journal of Sustainable Development and World Ecology
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
unsustainable practices
environmental emergencies
ecological overshoot
carrying capacity
societal evolution
Sustained Emergencies
Article
Text
ORIGINAL
cairns_sustained_emergencies.pdf
application/pdf
42928
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TEXT
cairns_sustained_emergencies.pdf.txt
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MD5
4
10919/24997
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/24997
2020-11-12 20:37:02.26
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1145382023-11-29T11:19:55Zcom_10919_23829com_10919_5553com_10919_103712com_10919_25796col_10919_23830col_10919_103713
Farzad, Roxanna
Ha, Anh D.
Aylward, Frank O.
2023-04-18T15:06:13Z
2023-04-18T15:06:13Z
2022-11
1021923
http://hdl.handle.net/10919/114538
https://doi.org/10.3389/fmicb.2022.1021923
13
36504832
1664-302X
Large double-stranded DNA viruses of the phylum Nucleocytoviricota, often referred to as "giant viruses," are ubiquitous members of marine ecosystems that are important agents of mortality for eukaryotic plankton. Although giant viruses are known to be prevalent in marine systems, their activities in oligotrophic ocean waters remain unclear. Oligotrophic gyres constitute the majority of the ocean and assessing viral activities in these regions is therefore critical for understanding overall marine microbial processes. In this study, we generated 11 metagenome-assembled genomes (MAGs) of giant viruses from samples previously collected from Station ALOHA in the North Pacific Subtropical Gyre. Phylogenetic analyses revealed that they belong to the orders Imitervirales (n =6), Algavirales (n =4), and Pimascovirales (n =1). Genome sizes ranged from similar to 119-574 kbp, and several of the genomes encoded predicted TCA cycle components, cytoskeletal proteins, collagen, rhodopsins, and proteins potentially involved in other cellular processes. Comparison with other marine metagenomes revealed that several have broad distribution across ocean basins and represent abundant viral constituents of pelagic surface waters. Our work sheds light on the diversity of giant viruses present in oligotrophic ocean waters across the globe.
Published version
application/pdf
en
Frontiers
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
giant viruses
marine viruses
station ALOHA
Prasinoviridae
Mesomimiviridae
Nucleocytoviricota
Diversity and genomics of giant viruses in the North Pacific Subtropical Gyre
Frontiers in Microbiology
Article - Refereed
Text
TEXT
fmicb-13-1021923.pdf.txt
fmicb-13-1021923.pdf.txt
Extracted text
text/plain
52851
https://vtechworks.lib.vt.edu/bitstreams/37f08144-5ef1-4bb9-ae9e-adfcacf5e39a/download
ecee4a51b15a28aa7e3ee8e7355aac28
MD5
2
THUMBNAIL
fmicb-13-1021923.pdf.jpg
fmicb-13-1021923.pdf.jpg
IM Thumbnail
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MD5
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ORIGINAL
fmicb-13-1021923.pdf
fmicb-13-1021923.pdf
Published version
application/pdf
3506215
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MD5
1
10919/114538
oai:vtechworks.lib.vt.edu:10919/114538
2023-11-29 11:19:55.491
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/766902023-11-29T19:07:23Zcom_10919_78629com_10919_78628com_10919_5540com_10919_23829com_10919_5553com_10919_111086com_10919_5532col_10919_78630col_10919_71752col_10919_23830col_10919_111087
Flint, Bonnie Fairbanks
Hawley, Dana M.
Alexander, Kathleen A.
Animal and Poultry Sciences
Biological Sciences
2017-03-28T17:31:55Z
2017-03-28T17:31:55Z
2016-07-25
http://hdl.handle.net/10919/76690
https://doi.org/10.1002/ece3.2343
6
16
Urbanization and other human modifications of the landscape may indirectly affect disease dynamics by altering host behavior in ways that influence pathogen transmission. Few opportunities arise to investigate behaviorally mediated effects of human habitat modification in natural host–pathogen systems, but we provide a potential example of this phenomenon in banded mongooses (Mungos mungo), a social mammal. Our banded mongoose study population in Botswana is endemically infected with a novel Mycobacterium tuberculosis complex pathogen, M. mungi, that primarily invades the mongoose host through the nasal planum and breaks in the skin. In this system, several study troops have access to human garbage sites and other modified landscapes for foraging. Banded mongooses in our study site (N = 4 troops, ~130 individuals) had significantly higher within-troop aggression levels when foraging in garbage compared to other foraging habitats. Second, monthly rates of aggression were a significant predictor of monthly number of injuries in troops. Finally, injured individuals had a 75% incidence of clinical tuberculosis (TB) compared to a 0% incidence in visibly uninjured mongooses during the study period. Our data suggest that mongoose troops that forage in garbage may be at greater risk of acquiring TB by incurring injuries that may allow for pathogen invasion. Our study suggests the need to consider the indirect effects of garbage on behavior and wildlife health when developing waste management approaches in human-modified areas.
application/pdf
en
Wiley
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Human-modified landscapes
provisioning
refuse
supplementation
urban wildlife
waste management
wildlife management
Do not feed the wildlife: associations between garbage use, aggression, and disease in banded mongooses (Mungos Mungo)
Ecology and Evolution
Article - Refereed
Text
ORIGINAL
AlexanderDoNotFeed2016.pdf
AlexanderDoNotFeed2016.pdf
application/pdf
116985
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MD5
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TEXT
AlexanderDoNotFeed2016.pdf.txt
AlexanderDoNotFeed2016.pdf.txt
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39606
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MD5
2
THUMBNAIL
AlexanderDoNotFeed2016.pdf.jpg
AlexanderDoNotFeed2016.pdf.jpg
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MD5
8
10919/76690
oai:vtechworks.lib.vt.edu:10919/76690
2023-11-29 19:07:23.003
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/491252021-07-16T12:56:01Zcom_10919_23829com_10919_5553col_10919_23830
Taylor, D. S.
Turner, Bruce J.
Davis, W. P.
Chapman, B. B.
Biological Sciences
2014-06-27T14:45:36Z
2014-06-27T14:45:36Z
2008-02
2014-06-26
D. Scott Taylor, Bruce J. Turner, William P. Davis, and Ben B. Chapman. "A Novel Terrestrial Fish Habitat inside Emergent Logs," The American Naturalist, Vol. 171, No. 2 (February 2008), pp. 263-266. DOI: 10.1086/524960
0003-0147
http://hdl.handle.net/10919/49125
http://www.jstor.org/stable/10.1086/524960
https://doi.org/10.1086/524960
Reports of new habitats for a major group of organisms are rare. Fishes display diverse adaptations for temporary (amphibious) existence on land, but to our knowledge, none have ever been reported regularly living inside emergent logs. Here, we show that the mangrove killifish, Kryptolebias marmoratus, a species previously known to emerse (leave the water) regularly, is now known to emerse and aggregate in large numbers inside decaying mangrove logs that have been "galleried" by terrestrial insects. This behavior has now been documented in both Belize, Central America, and Florida, U.S.A., populations and represents the first known case of fishes entering terrestrial woody material. The dense packing of fish in the narrow log galleries may imply a novel social context in which intraspecific aggressive behaviors are reduced, possibly mediated by the physiological limitations imposed within this restrictive habitat.
CCRE contribution 809
Jeffress Memorial Trust
University of Leeds
Nuffield Foundation
en_US
University of Chicago Press
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
red mangrove
kryptolebias marmoratus
emersion
amphibious fish
rivulus-marmoratus
hermaphroditic fish
ecology
evolutionary biology
Natural history note - A novel terrestrial fish habitat inside emergent logs
American Naturalist
Article - Refereed
ORIGINAL
524960.pdf
524960.pdf
Main article
application/pdf
486647
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MD5
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TEXT
524960.pdf.txt
524960.pdf.txt
Extracted text
text/plain
16609
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/8ec6cdda-0b21-4a2c-afa2-cdc57c19bb62/download
7be11d0a277556758203c28890df3922
MD5
2
THUMBNAIL
524960.pdf.jpg
524960.pdf.jpg
IM Thumbnail
image/jpeg
4978
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2ff119263547a77fc0e6fc592e5fd443
MD5
4
10919/49125
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/49125
2021-07-16 12:56:01.099
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/820522024-03-12T15:58:33Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553com_10919_25796col_10919_70873col_10919_23830col_10919_25797
Briggs, Amy G.
Hughes, Lee E.
Brennan, Robert E.
Buchner, John
Horak, Rachel E. A.
Katz Amburn, D. Sue
McDonald, Ann H.
Primm, Todd P.
Smith, Ann C.
Stevens, Ann M.
Yung, Sunny B.
Paustian, Timothy D.
Biological Sciences
Fralin Life Sciences Institute
2018-02-09T19:05:19Z
2018-02-09T19:05:19Z
2017-10
http://hdl.handle.net/10919/82052
https://doi.org/10.1128/jmbe.v18i3.1319
18
3
American Society for Microbiology
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
Concept inventory development reveals common student misconceptions about microbiology
Journal of Microbiology and Biology Education
Article - Refereed
Published version
/Virginia Tech
/Virginia Tech/All T&R Faculty
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Science/COS T&R Faculty
/Virginia Tech/University Research Institutes
/Virginia Tech/University Research Institutes/Fralin Life Sciences
/Virginia Tech/University Research Institutes/Fralin Life Sciences/Fralin Affiliated Faculty
ORIGINAL
JMBEMisconceptions.pdf
Publisher's Version
application/pdf
568419
https://vtechworks.lib.vt.edu/bitstreams/724bebf0-e344-4020-a3db-72f2e587f1e4/download
5ece208c59f7c83308774962cc507a46
MD5
1
LICENSE
VTUL_Distribution_License_2016_05_09.pdf
application/pdf
18524
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MD5
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TEXT
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JMBEMisconceptions.pdf.txt
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MD5
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MD5
8
10919/82052
oai:vtechworks.lib.vt.edu:10919/82052
2024-03-12 15:58:33.305
http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
open.access
https://vtechworks.lib.vt.edu
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vtechworks@vt.edu
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oai:vtechworks.lib.vt.edu:10919/1163362023-09-27T07:12:46Zcom_10919_23829com_10919_5553col_10919_23830
Nelson, Annika S.
Gelambi, Mariana
Morales-M., Estefania
Whitehead, Susan R.
2023-09-26T13:44:53Z
2023-09-26T13:44:53Z
2023-05
0012-9658
http://hdl.handle.net/10919/116336
https://doi.org/10.1002/ecy.4032
104
5
36932996
1939-9170
Plant secondary metabolites are key mechanistic drivers of species interactions. These metabolites have primarily been studied for their role in defense, but they can also have important consequences for mutualisms, including seed dispersal. Although the primary function of fleshy fruits is to attract seed-dispersing animals, fruits often contain complex mixtures of toxic or deterrent secondary metabolites that can reduce the quantity or quality of seed dispersal mutualisms. Furthermore, because seeds are often dispersed across multiple stages by several dispersers, the net consequences of fruit secondary metabolites for the effectiveness of seed dispersal and ultimately plant fitness are poorly understood. Here, we tested the effects of amides, nitrogen-based defensive compounds common in fruits of the neotropical plant genus Piper (Piperaceae), on seed dispersal effectiveness (SDE) by ants, which are common secondary seed dispersers. We experimentally added amide extracts to Piper fruits both in the field and lab, finding that amides reduced the quantity of secondary seed dispersal by reducing ant recruitment (87%) and fruit removal rates (58% and 66% in the field and lab, respectively). Moreover, amides not only reduced dispersal quantity but also altered seed dispersal quality by shifting the community composition of recruiting ants (notably by reducing the recruitment of the most effective disperser by 90% but having no detectable effect on the recruitment of a cheater species that removes fruit pulp without dispersing seeds). Although amides did not affect the distance ants initially carried seeds, they altered the quality of seed dispersal by reducing the likelihood of ants cleaning seeds (67%) and increasing their likelihood of ants redispersing seeds outside of the nest (200%). Overall, these results demonstrate that secondary metabolites can alter the effectiveness of plant mutualisms, by both reducing mutualism quantity and altering mutualism quality through multiple mechanisms. These findings present a critical step in understanding the factors mediating the outcomes of seed dispersal and, more broadly, demonstrate the importance of considering how defensive secondary metabolites influence the outcomes of mutualisms surrounding plants.
National Science Foundation [1856776]
Published version
National Science Foundation,Grant/Award Number: 1856776
application/pdf
en
Wiley
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
ants
chemical ecology
fruit defense
mutualism
plant secondary metabolites
seed dispersal effectiveness
seed dispersal quality
seed dispersal quantity
Fruit secondary metabolites alter the quantity and quality of a seed dispersal mutualism
Ecology
Article - Refereed
Text
ORIGINAL
Ecology - 2023 - Nelson.pdf
Ecology - 2023 - Nelson.pdf
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Creative Commons Attribution 4.0 International
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oai:vtechworks.lib.vt.edu:10919/736292024-03-13T14:09:53Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553col_10919_70873col_10919_23830
Gerard, Claude
Tyson, John J.
Coudreuse, Damien
Novak, Bela
Biological Sciences
2016-12-09T21:33:39Z
2016-12-09T21:33:39Z
2015-02-01
1553-734X
http://hdl.handle.net/10919/73629
https://doi.org/10.1371/journal.pcbi.1004056
11
2
In present-day eukaryotes, the cell division cycle is controlled by a complex network of interacting proteins, including members of the cyclin and cyclin-dependent protein kinase (Cdk) families, and the Anaphase Promoting Complex (APC). Successful progression through the cell cycle depends on precise, temporally ordered regulation of the functions of these proteins. In light of this complexity, it is surprising that in fission yeast, a minimal Cdk network consisting of a single cyclin-Cdk fusion protein can control DNA synthesis and mitosis in a manner that is indistinguishable from wild type. To improve our understanding of the cell cycle regulatory network, we built and analysed a mathematical model of the molecular interactions controlling the G1/S and G2/M transitions in these minimal cells. The model accounts for all observed properties of yeast strains operating with the fusion protein. Importantly, coupling the model’s predictions with experimental analysis of alternative minimal cells, we uncover an explanation for the unexpected fact that elimination of inhibitory phosphorylation of Cdk is benign in these strains while it strongly affects normal cells. Furthermore, in the strain without inhibitory phosphorylation of the fusion protein, the distribution of cell size at division is unusually broad, an observation that is accounted for by stochastic simulations of the model. Our approach provides novel insights into the organization and quantitative regulation of wild type cell cycle progression. In particular, it leads us to propose a new mechanistic model for the phenomenon of mitotic catastrophe, relying on a combination of unregulated, multi-cyclin-dependent Cdk activities.
Published version
? - ? (27) page(s)
English
PLOS
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000352081000022&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Biochemical Research Methods
Mathematical & Computational Biology
Biochemistry & Molecular Biology
FISSION YEAST
S-PHASE
DNA-REPLICATION
PROTEIN-KINASE
SCHIZOSACCHAROMYCES-POMBE
MULTISITE PHOSPHORYLATION
TYROSINE PHOSPHORYLATION
GENE-EXPRESSION
MITOTIC CONTROL
FEEDBACK LOOPS
Cell Cycle Control by a Minimal Cdk Network
PLOS Computational Biology
Article - Refereed
/Virginia Tech
/Virginia Tech/All T&R Faculty
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Science/COS T&R Faculty
/Virginia Tech/University Distinguished Professors
ORIGINAL
Cell cycle control by a minimal Cdk network.pdf
Publisher's Version
application/pdf
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oai:vtechworks.lib.vt.edu:10919/250862020-11-12T20:37:03Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:32Z
2014-01-23T02:36:32Z
2006
http://hdl.handle.net/10919/25086
http://www.johncairns.net/Commentaries/securitycommentary.pdf
The fossil record shows that all Earth s species have been transient. Yet humankind espouses sustainable development (which is based on continued human occupancy of the planet) despite the probability that Earth may last another 15 billion years. Is this expectation hubris, denial of scientific evidence, or failure to engage in any fundamental change in social norms? If unsustainable practices continue, leaving a habitable planet for posterity will not be possible. The most crucial determinant of human destiny on the planet is the continual health and integrity of the biospheric life support system, which has produced conditions (e.g., atmosphere gas balance) favorable to Homo sapiens for 160,000 years. If major alterations occur in the function of the biospheric life support system, present human social systems will be in disequilibrium, and even human survival will be in doubt.
en_US
Virginia Tech
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
biospheric life support system
sustainable development
sustainablility
human survival
Human Density and the Biospheric Life Support System
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2020-11-12 20:37:03.248
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oai:vtechworks.lib.vt.edu:10919/749082023-11-29T19:07:29Zcom_10919_5com_10919_25799com_10919_8195com_10919_5540com_10919_23829com_10919_5553com_10919_25796com_10919_24213col_10919_70873col_10919_18629col_10919_71752col_10919_23830col_10919_25797col_10919_24334
Wu, Xiaowei
Sun, Ming-an
Zhu, Hongxiao
Xie, Hehuang
Statistics
2017-02-02T22:54:04Z
2017-02-02T22:54:04Z
2015-01-16
1471-2105
http://hdl.handle.net/10919/74908
https://doi.org/10.1186/s12859-014-0439-2
16
Background: With recent development in sequencing technology, a large number of genome-wide DNA methylation studies have generated massive amounts of bisulfite sequencing data. The analysis of DNA methylation patterns helps researchers understand epigenetic regulatory mechanisms. Highly variable methylation patterns reflect stochastic fluctuations in DNA methylation, whereas well-structured methylation patterns imply deterministic methylation events. Among these methylation patterns, bipolar patterns are important as they may originate from allele-specific methylation (ASM) or cell-specific methylation (CSM).
Results: Utilizing nonparametric Bayesian clustering followed by hypothesis testing, we have developed a novel statistical approach to identify bipolar methylated genomic regions in bisulfite sequencing data. Simulation studies demonstrate that the proposed method achieves good performance in terms of specificity and sensitivity. We used the method to analyze data from mouse brain and human blood methylomes. The bipolar methylated segments detected are found highly consistent with the differentially methylated regions identified by using purified cell subsets.
Conclusions: Bipolar DNA methylation often indicates epigenetic heterogeneity caused by ASM or CSM. With allele-specific events filtered out or appropriately taken into account, our proposed approach sheds light on the identification of cell-specific genes/pathways under strong epigenetic control in a heterogeneous cell population.
Published version
12 page(s)
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en
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http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000348816000001&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
The Author(s)
Biochemical Research Methods
Biotechnology & Applied Microbiology
Mathematical & Computational Biology
Biochemistry & Molecular Biology
DNA methylation
Epigenetics
Nonparametric Bayesian
BISULFITE SEQUENCING DATA
DNA METHYLATION
CAENORHABDITIS-ELEGANS
EPIGENOMIC ANALYSIS
HUMAN-NEUTROPHILS
ALU REPEATS
PATTERNS
ORIGIN
DIFFERENTIATION
SPECIFICITY
Nonparametric Bayesian clustering to detect bipolar methylated genomic loci
BMC Bioinformatics
Article - Refereed
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oai:vtechworks.lib.vt.edu:10919/971052021-10-08T16:43:29Zcom_10919_78629com_10919_78628com_10919_8195com_10919_25799com_10919_23829com_10919_5553col_10919_78630col_10919_78882col_10919_23830
Szwetkowski, Kyle J.
Falkinham, Joseph O. III
Biological Sciences
2020-03-02T13:29:44Z
2020-03-02T13:29:44Z
2020-02-22
2020-03-02T12:41:52Z
Szwetkowski, K.J.; Falkinham, J.O., III. Methylobacterium spp. as Emerging Opportunistic Premise Plumbing Pathogens. Pathogens 2020, 9, 149.
http://hdl.handle.net/10919/97105
https://doi.org/10.3390/pathogens9020149
<i>Methylobacterium</i> spp. are emerging opportunistic premise plumbing pathogens. Human infections linked to premise plumbing provide evidence of their routes of infection. Cells of a collection of representative strains of different <i>Methylobacterium</i> species were tested for hydrophobicity by contact angle, adherence and biofilm formation on different plumbing materials, and temperature tolerance (50–60 °C); characteristics shared by OPPPs. <i>Methylobacterium</i> spp. strains were shown to grow in drinking water, have high cell-surface hydrophobicity, adhere to pipe surface materials, form biofilms, and survive exposure to high (60° C) temperatures. It can be concluded that <i>Methylobacterium</i> spp. strains share traits in common with other opportunistic premise plumbing pathogens (OPPPs).
Published version
application/pdf
en
MDPI
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
opportunistic pathogen
adherence
contact angle
biofilm
temperature susceptibility
Methylobacterium spp. as Emerging Opportunistic Premise Plumbing Pathogens
Pathogens
Article - Refereed
Text
ORIGINAL
pathogens-09-00149.pdf
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2021-10-08 16:43:29.814
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/895412020-10-22T03:35:25Zcom_10919_23829com_10919_5553col_10919_23830
Hopkins, William A.
Fallon, Jesse A.
Beck, Michelle L.
Coe, Brittney H.
Jachowski, Catherine M.B.
Biological Sciences
2019-05-16T17:52:39Z
2019-05-16T17:52:39Z
2016
http://hdl.handle.net/10919/89541
https://doi.org/10.1093/conphys/cow002
4
Disease is among the leading causes of the global decline in amphibian populations. In North America, parasites and pathogens are among the factors implicated in precipitous population declines of the giant hellbender salamander (Cryptobranchus alleganiensis), but the incidence of infections and the responses of hellbenders to infections remain poorly studied. Here, we document the prevalence of leech and trypanosome infections in a wild population of eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) and describe haematological and immunological characteristics of hellbenders harbouring these infections. We hypothesized that hellbenders parasitized by trypanosomes would be anaemic, that individuals infected with either or both parasites would exhibit shifts in white blood cell counts and that hellbenders infected with leeches would exhibit altered plasma bactericidal capacity. We found that 24 and 68% of hellbenders in our sample population were infected with leeches and trypanosomes, respectively, and 20% were co-infected with both parasites. We found no evidence suggestive of anaemia among infected individuals. However, hellbenders infected with either or both parasites exhibited marked shifts in circulating white blood cells that were consistent with predictable responses to parasitic infection. Additionally, we found that hellbenders harbouring leeches had much higher plasma bactericidal capacity than individuals without leeches, and we offer multiple potential mechanistic explanations for this observation. We also found evidence that cellular and serological immune responses to parasites were less robust in juvenile than adult hellbenders. This finding warrants further investigation in light of the demographic characteristics, specifically the scarcity of juvenile age classes, of hellbender populations where disease is a possible contributor to declines. Finally, we describe two methodological advances that will improve future studies seeking to diagnose trypanosome infections and to test the bactericidal capacity of hellbenders and perhaps other amphibians. Our study provides fundamental insights into how hellbenders respond physiologically to endo- and ectoparasites, which could ultimately prove useful for their conservation.
Virginia Department of Game and Inland Fisheries
Fralin Life Science Institute
en_US
Oxford University Press
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Amphibian
anaemia
co-infection
leech
neutrophil-to-lymphocyte ratio
trypanosome
Haematological and immunological characteristics of eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) infected and co-infected with endo- and ectoparasites
Conservation Physiology
Article - Refereed
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oai:vtechworks.lib.vt.edu:10919/748822023-11-29T19:07:31Zcom_10919_5com_10919_25799com_10919_5540com_10919_23829com_10919_5553com_10919_25796col_10919_70873col_10919_71752col_10919_23830col_10919_25797
Rodrigues, Richard R.
Pineda, Rosana P.
Barney, Jacob
Nilsen, Erik T.
Barrett, John E.
Williams, Mark A.
Biological Sciences
Fralin Life Sciences Institute
2017-02-01T19:11:11Z
2017-02-01T19:11:11Z
2015-10-27
1932-6203
http://hdl.handle.net/10919/74882
https://doi.org/10.1371/journal.pone.0141424
10
10
The importance of plant-microbe associations for the invasion of plant species have not been often tested under field conditions. The research sought to determine patterns of change in microbial communities associated with the establishment of invasive plants with different taxonomic and phenetic traits. Three independent locations in Virginia, USA were selected. One site was invaded by a grass (Microstegium vimineum), another by a shrub (Rhamnus davurica), and the third by a tree (Ailanthus altissima). The native vegetation from these sites was used as reference. 16S rRNA and ITS regions were sequenced to study root-zone bacterial and fungal communities, respectively, in invaded and non-invaded samples and analyzed using Quantitative Insights Into Microbial Ecology (QIIME). Though root-zone microbial community structure initially differed across locations, plant invasion shifted communities in similar ways. Indicator species analysis revealed that Operational Taxonomic Units (OTUs) closely related to Proteobacteria, Acidobacteria, Actinobacteria, and Ascomycota increased in abundance due to plant invasions. The Hyphomonadaceae family in the Rhodobacterales order and ammonia-oxidizing Nitrospirae phylum showed greater relative abundance in the invaded root-zone soils. Hyphomicrobiaceae, another bacterial family within the phyla Proteobacteria increased as a result of plant invasion, but the effect associated most strongly with root-zones ofM. vimineum and R. davurica. Functional analysis using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) showed bacteria responsible for nitrogen cycling in soil increased in relative abundance in association with plant invasion. In agreement with phylogenetic and functional analyses, greater turnover of ammonium and nitrate was associated with plant invasion. Overall, bacterial and fungal communities changed congruently across plant invaders, and support the hypothesis that nitrogen cycling bacteria and functions are important factors in plant invasions. Whether the changes in microbial communities are driven by direct plant microbial interactions or a result of plant-driven changes in soil properties remains to be determined.
Published version
? - ? (19) page(s)
application/pdf
English
PLOS
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000363804200065&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
ailanthus-altissima
soil
nitrogen
ecosystems
sequences
fungi
tree
identification
metaanalysis
hypotheses
Plant Invasions Associated with Change in Root-Zone Microbial Community Structure and Diversity
PLOS ONE
Article - Refereed
Text
/Virginia Tech
/Virginia Tech/Agriculture & Life Sciences
/Virginia Tech/Agriculture & Life Sciences/CALS T&R Faculty
/Virginia Tech/Agriculture & Life Sciences/Horticulture
/Virginia Tech/Agriculture & Life Sciences/Plant Pathology, Physiology, & Weed Science
/Virginia Tech/All T&R Faculty
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Science/COS T&R Faculty
ORIGINAL
Plant Invasions Associated with Change in Root-Zone Microbial Community Structure and Diversity.pdf
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oai:vtechworks.lib.vt.edu:10919/1176882024-01-26T03:00:59Zcom_10919_23829com_10919_5553com_10919_24259com_10919_5559com_10919_25796com_10919_111086com_10919_5532com_10919_84995col_10919_23830col_10919_24342col_10919_25797col_10919_111087col_10919_84996
Farrell, Kayla
Auerbach, Aubrey
Musaus, Madeline
Navabpour, Shaghayegh
Liu, Catherine
Lin, Yu
Xie, Hehuang
Jarome, Timothy J.
2024-01-25T20:43:51Z
2024-01-25T20:43:51Z
2024-01
https://hdl.handle.net/10919/117688
https://doi.org/10.1523/JNEUROSCI.1453-23.2023
44
4
Memory formation requires coordinated control of gene expression, protein synthesis, and ubiquitin–proteasome system (UPS)-mediated protein degradation. The catalytic component of the UPS, the 26S proteasome, contains a 20S catalytic core surrounded by two 19S regulatory caps, and phosphorylation of the 19S cap regulatory subunit RPT6 at serine 120 (pRPT6-S120) has been widely implicated in controlling activity-dependent increases in proteasome activity. Recently, RPT6 was also shown to act outside the proteasome where it has a transcription factor-like role in the hippocampus during memory formation. However, little is known about the proteasome-independent function of “free” RPT6 in the brain or during memory formation and whether phosphorylation of S120 is required for this transcriptional control function. Here, we used RNA-sequencing along with novel genetic approaches and biochemical, molecular, and behavioral assays to test the hypothesis that pRPT6-S120 functions independently of the proteasome to bind DNA and regulate gene expression during memory formation. RNA-sequencing following siRNA-mediated knockdown of free RPT6 revealed 46 gene targets in the dorsal hippocampus of male rats following fear conditioning, where RPT6 was involved in transcriptional activation and repression. Through CRISPR-dCas9-mediated artificial placement of RPT6 at a target gene, we found that RPT6 DNA binding alone may be important for altering gene expression following learning. Further, CRISPR-dCas13-mediated conversion of S120 to glycine on RPT6 revealed that phosphorylation at S120 is necessary for RPT6 to bind DNA and properly regulate transcription during memory formation. Together, we reveal a novel function for phosphorylation of RPT6 in controlling gene transcription during memory formation.
This work was supported by National Institutes of Health grants MH122414, MH131587, MH120498, MH120569, MH123742, AG071523, and AG079292 to T.J.J.
application/pdf
en
Society for Neuroscience
CRISPR
Hippocampus
memory
proteasome
RPT6
transcription
Phosphorylation of RPT6 Controls Its Ability to Bind DNA and Regulate Gene Expression in the Hippocampus of Male Rats during Memory Formation
Journal of Neuroscience
Article - Refereed
Text
ORIGINAL
Farrell et al. 2023 J Neurosci.pdf
Farrell et al. 2023 J Neurosci.pdf
Accepted version
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oai:vtechworks.lib.vt.edu:10919/117688
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vtechworks@vt.edu
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oai:vtechworks.lib.vt.edu:10919/853812023-04-14T17:49:42Zcom_10919_23829com_10919_5553col_10919_23830
Hu, Yuanjie
Ru, Ning
Xiao, Huasheng
Chaturbedi, Abhishek
Hoa, Neil T.
Tian, Xiao-Jun
Zhang, Hang
Ke, Chao
Yan, Fengrong
Nelson, Jodi
Li, Zhenzhi
Gramer, Robert
Yu, Liping
Siegel, Eric
Zhang, Xiaona
Jia, Zhenyu
Jadus, Martin R.
Limoli, Charles L.
Linskey, Mark E.
Xing, Jianhua
Zhou, Yi-Hong
Biological Sciences
2018-10-16T16:41:24Z
2018-10-16T16:41:24Z
2013-09-25
e80898
http://hdl.handle.net/10919/85381
https://doi.org/10.1371/journal.pone.0080898
8
11
24282558
1932-6203
Aneuploidy with chromosome instability is a cancer hallmark. We studied chromosome 7 (Chr7) copy number variation (CNV) in gliomas and in primary cultures derived from them. We found tumor heterogeneity with cells having Chr7-CNV commonly occurs in gliomas, with a higher percentage of cells in high-grade gliomas carrying more than 2 copies of Chr7, as compared to low-grade gliomas. Interestingly, all Chr7-aneuploid cell types in the parental culture of established glioma cell lines reappeared in single-cell-derived subcultures. We then characterized the biology of three syngeneic glioma cultures dominated by different Chr7-aneuploid cell types. We found phenotypic divergence for cells following Chr7 mis-segregation, which benefited overall tumor growth in vitro and in vivo. Mathematical modeling suggested the involvement of chromosome instability and interactions among cell subpopulations in restoring the optimal equilibrium of tumor cell types. Both our experimental data and mathematical modeling demonstrated that the complexity of tumor heterogeneity could be enhanced by the existence of chromosomes with structural abnormality, in addition to their mis-segregations. Overall, our findings show, for the first time, the involvement of chromosome instability in maintaining tumor heterogeneity, which underlies the enhanced growth, persistence and treatment resistance of cancers.
Published version
application/pdf
en
PLOS
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Tumor-Specific Chromosome Mis-Segregation Controls Cancer Plasticity by Maintaining Tumor Heterogeneity
PLOS ONE
Article - Refereed
Text
ORIGINAL
journal.pone.0080898.PDF
journal.pone.0080898.PDF
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oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/85381
2023-04-14 17:49:42.042
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/932882020-10-22T03:36:55Zcom_10919_23829com_10919_5553col_10919_23830
Cheng, Keqiang
Geng, Shuo
Yuan, Ruoxi
Diao, Na
Upchurch, Zachary
Li, Liwu
Biological Sciences
2019-08-29T12:25:41Z
2019-08-29T12:25:41Z
2015-03-06
http://hdl.handle.net/10919/93288
https://doi.org/10.1016/j.ebiom.2015.03.001
2
Sepsis mortality varies dramatically in individuals of variable immune conditions, with poorly defined mechanisms. This phenomenon complements the hypothesis that innate immunity may adopt rudimentary memory, as demonstrated in vitro with endotoxin priming and tolerance in cultured monocytes. However, previous in vivo studies only examined the protective effect of endotoxin tolerance in the context of sepsis. In sharp contrast, we report herein that pre-conditioning with super-low or low dose endotoxin lipopolysaccharide (LPS) cause strikingly opposite survival outcomes. Mice pre-conditioned with super-low dose LPS experienced severe tissue damage, inflammation, increased bacterial load in circulation, and elevatedmortalitywhen theywere subjected to cecal-ligation and puncture (CLP). This is in contrast to thewell-reported protective phenomenon with CLPmice pre-conditioned with lowdose LPS.Mechanistically,we demonstrated that super-lowand lowdose LPS differentially modulate the formation of neutrophil extracellular trap (NET) in neutrophils. Instead of increased ERK activation and NET formation in neutrophils pre-conditioned with low dose LPS, we observed significantly reduced ERK activation and compromised NET generation in neutrophils pre-conditioned with super-low dose LPS. Collectively, our findings reveal amechanismpotentially responsible for the dynamic programming of innate immunity in vivo as it relates to sepsis risks.
This work is supported in part by grants from the National Institute of Health R01 HL115835 and R56 AI108264 to L.L.
10 pages
application/pdf
en
Elsevier
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
Innate immunity
Pre-conditioning dynamics
Neutrophil
Polymicrobial sepsis
Super-low dose endotoxin
Super-low Dose Endotoxin Pre-conditioning Exacerbates Sepsis Mortality
EBioMedicine
Article - Refereed
Text
ORIGINAL
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oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/93288
2020-10-22 03:36:55.388
http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1115772022-08-23T07:13:15Zcom_10919_23829com_10919_5553col_10919_23830
Zhang, Tongli
Tyson, John J.
2022-08-22T13:47:29Z
2022-08-22T13:47:29Z
2022-02
1567-567X
http://hdl.handle.net/10919/111577
https://doi.org/10.1007/s10928-021-09798-1
49
1
34985622
1573-8744
Individual biological organisms are characterized by daunting heterogeneity, which precludes describing or understanding populations of 'patients' with a single mathematical model. Recently, the field of quantitative systems pharmacology (QSP) has adopted the notion of virtual patients (VPs) to cope with this challenge. A typical population of VPs represents the behavior of a heterogeneous patient population with a distribution of parameter values over a mathematical model of fixed structure. Though this notion of VPs is a powerful tool to describe patients' heterogeneity, the analysis and understanding of these VPs present new challenges to systems pharmacologists. Here, using a model of the hypothalamic-pituitary-adrenal axis, we show that an integrated pipeline that combines machine learning (ML) and bifurcation analysis can be used to effectively and efficiently analyse the behaviors observed in populations of VPs. Compared with local sensitivity analyses, ML allows us to capture and analyse the contributions of simultaneous changes of multiple model parameters. Following up with bifurcation analysis, we are able to provide rigorous mechanistic insight regarding the influences of ML-identified parameters on the dynamical system's behaviors. In this work, we illustrate the utility of this pipeline and suggest that its wider adoption will facilitate the use of VPs in the practice of systems pharmacology.
[1016183 ARMY W911NF-20-1-0192]
Published version
The work was supported by Grant 1016183 ARMY W911NF-20-1-0192 to TZ.
application/pdf
en
Springer/Plenum Publishers
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Quantitative systems pharmacology
Virtual patients
Machine learning
Bifurcation analysis
Nonlinear dynamics
Hypothalamic-pituitary-adrenal axis
Understanding virtual patients efficiently and rigorously by combining machine learning with dynamical modelling
Journal of Pharmacokinetics and Pharmacodynamics
Article - Refereed
Text
ORIGINAL
s10928-021-09798-1.pdf
s10928-021-09798-1.pdf
Published version
application/pdf
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3
10919/111577
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/111577
2022-08-23 07:13:15.863
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/943532020-10-22T03:35:28Zcom_10919_23829com_10919_5553col_10919_23830
Duong, Duy An
Stevens, Ann M.
Jensen, Roderick V.
Biological Sciences
2019-10-04T14:40:51Z
2019-10-04T14:40:51Z
2017-06
2576-098X
e00435-17
http://hdl.handle.net/10919/94353
https://doi.org/10.1128/genomeA.00435-17
5
22
28572317
The phytopathogen Pantoea stewartii subsp. stewartii DC283 causes Stewart's wilt disease in corn after transmission from the corn flea beetle insect vector. Here, we report that the complete annotated genome of P. stewartii DC283 has been fully assembled into one circular chromosome, 10 circular plasmids, and one linear phage.
Fralin Life Science Institute; Biological Sciences Department, Virginia Tech
This work was funded by the Fralin Life Science Institute and Biological Sciences Department, Virginia Tech.
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Complete Genome Assembly of Pantoea stewartii subsp. stewartii DC283, a Corn Pathogen
Microbiology Resource Announcements
Article - Refereed
Text
StillImage
ORIGINAL
Microbiology Resource Announcements-2017-Duong-e00435-17.full.pdf
Microbiology Resource Announcements-2017-Duong-e00435-17.full.pdf
application/pdf
121800
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/ca360bb4-3c57-40eb-9669-e5eed733d12b/download
7567b1fc181726781389704aed663ed5
MD5
1
TEXT
Microbiology Resource Announcements-2017-Duong-e00435-17.full.pdf.txt
Microbiology Resource Announcements-2017-Duong-e00435-17.full.pdf.txt
Extracted text
text/plain
7488
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/a4368755-ca12-4176-a349-88167ebfefd0/download
b2d79cb1415c517629e2a5b36077f39e
MD5
2
THUMBNAIL
Microbiology Resource Announcements-2017-Duong-e00435-17.full.pdf.jpg
Microbiology Resource Announcements-2017-Duong-e00435-17.full.pdf.jpg
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MD5
4
10919/94353
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/94353
2020-10-22 03:35:28.044
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/862682020-10-22T03:26:47Zcom_10919_23829com_10919_5553col_10919_23830
Levin, Iris I.
Fosdick, Bailey K.
Tsunekage, Toshi
Aberle, Matthew A.
Burns, Christine M. Bergeon
Hund, Amanda K.
Safran, Rebecca J.
Biological Sciences
2018-12-07T20:25:04Z
2018-12-07T20:25:04Z
2018-10-19
2045-2322
15533
http://hdl.handle.net/10919/86268
https://doi.org/10.1038/s41598-018-33948-0
8
30341385
Animals use morphological signals such as ornamental traits or weaponry to mediate social interactions, and the extent of signal trait elaboration is often positively associated with reproductive success. By demonstrating relationships between signal traits and fitness, researchers often make inferences about how behaviour operates to shape those outcomes. However, detailed information about fine-scale individual behaviour, and its physiological basis, can be difficult to obtain. Here we show that experimental manipulations to exaggerate a signal trait (plumage colour) and concomitant changes in testosterone and stress-induced corticosterone levels altered social interactivity between manipulated males and their social mates. On average, darkened males did not have higher levels of interactivity than unmanipulated males; however, males who experienced a greater shift in colour (pale to dark), a larger, positive change in testosterone levels, and a dampened stress-induced corticosterone response had a larger increase in the number of interactions with their social mate post-manipulation compared to pre-manipulation. This work provides new insights into the integration and real-time flexibility of multivariate phenotypes and direct evidence for the role of social interactions in pair bond maintenance.
University of Colorado Innovative Seed Grant; National Science Foundation Postdoctoral Research Fellowship [DBI-1306059]
We thank J. Burt for assistance with the tags, S. Turbek, and T. Albrecht for assistance in the field, and Colorado Horse Rescue for access to their barn. A University of Colorado Innovative Seed Grant provided funding for this research and I. Levin was supported by a National Science Foundation Postdoctoral Research Fellowship (DBI-1306059).
7 pages
application/pdf
en_US
Springer Nature
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
american barn swallows
plumage color
sexual selection
hirundo-rustica
stress
corticosterone
testosterone
systems
length
junco
Experimental manipulation of a signal trait reveals complex phenotype-behaviour coordination
Scientific Reports
Article - Refereed
Text
ORIGINAL
s41598-018-33948-0.pdf
s41598-018-33948-0.pdf
application/pdf
1089507
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/08b6f9d9-c834-43b4-b8ec-d1e058dacb99/download
308482d38c808c9a0d7357e9f294a0ce
MD5
1
TEXT
s41598-018-33948-0.pdf.txt
s41598-018-33948-0.pdf.txt
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40602
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/830160e8-5d02-4edb-a17c-f83d3b0add4f/download
dd89474f445c3b98d16baa2ede27a178
MD5
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db206b291dd15ff1c0c1f8e6090233e2
MD5
4
10919/86268
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/86268
2020-10-22 03:26:47.306
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1010402021-12-16T15:39:58Zcom_10919_23829com_10919_5553com_10919_24216com_10919_5539com_10919_24214com_10919_23261col_10919_23830col_10919_24286col_10919_24288col_10919_23262
Moarefian, Maryam
Davalos, Rafael V.
Tafti, Danesh K.
Achenie, Luke E. K.
Jones, Caroline N.
Mechanical Engineering
Biomedical Engineering and Mechanics
Chemical Engineering
Biological Sciences
2020-12-08T14:10:33Z
2020-12-08T14:10:33Z
2020-09-21
1473-0197
http://hdl.handle.net/10919/101040
https://doi.org/10.1039/d0lc00602e
20
18
32869052
1473-0189
Iontophoresis employs low-intensity electrical voltage and continuous constant current to direct a charged drug into a tissue. Iontophoretic drug delivery has recently been used as a novel method for cancer treatment in vivo. There is an urgent need to precisely model the low-intensity electric fields in cell culture systems to optimize iontophoretic drug delivery to tumors. Here, we present an iontophoresis-on-chip (IOC) platform to precisely quantify carboplatin drug delivery and its corresponding anti-cancer efficacy under various voltages and currents. In this study, we use an in vitro heparin-based hydrogel microfluidic device to model the movement of a charged drug across an extracellular matrix (ECM) and in MDA-MB231 triple-negative breast cancer (TNBC) cells. Transport of the drug through the hydrogel was modeled based on diffusion and electrophoresis of charged drug molecules in the direction of an oppositely charged electrode. The drug concentration in the tumor extracellular matrix was computed using finite element modeling of transient drug transport in the heparin-based hydrogel. The model predictions were then validated using the IOC platform by comparing the predicted concentration of a fluorescent cationic dye (Alexa Fluor 594 (R)) to the actual concentration in the microfluidic device. Alexa Fluor 594 (R) was used because it has a molecular weight close to paclitaxel, the gold standard drug for treating TNBC, and carboplatin. Our results demonstrated that a 50 mV DC electric field and a 3 mA electrical current significantly increased drug delivery and tumor cell death by 48.12% +/- 14.33 and 39.13% +/- 12.86, respectively (n = 3, p-value <0.05). The IOC platform and mathematical drug delivery model of iontophoresis are promising tools for precise delivery of chemotherapeutic drugs into solid tumors. Further improvements to the IOC platform can be made by adding a layer of epidermal cells to model the skin.
Department of Biological Sciences at Virginia Tech; National Institute of General Medical Sciences of the National Institutes of HealthUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of General Medical Sciences (NIGMS) [R35GM133610]; VT-Initiative for Maximizing Student Development (IMSD) (NIGMS) [2R25GM072767-05A1]; Department of Mechanical Engineering at Virginia Tech.
Research materials and equipment for this study were funded by CNJ startup funds from the Department of Biological Sciences at Virginia Tech and The National Institute of General Medical Sciences of the National Institutes of Health under award number R35GM133610. MM was supported by the VT-Initiative for Maximizing Student Development (IMSD) (NIGMS 2R25GM072767-05A1) and the Department of Mechanical Engineering at Virginia Tech.
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Modeling iontophoretic drug delivery in a microfluidic device
Lab On A Chip
Article - Refereed
Text
StillImage
TEXT
d0lc00602e.pdf.txt
d0lc00602e.pdf.txt
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text/plain
54926
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/50937ecb-aa41-43c8-a190-ef744b16bad3/download
6284eb1073f025f3d7cf74d229af1b2d
MD5
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11869e3b1f4940b6627e6c56fa308bea
MD5
3
ORIGINAL
d0lc00602e.pdf
d0lc00602e.pdf
application/pdf
5232055
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/a839f87b-735d-457f-90c1-f101713bf8ff/download
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MD5
1
10919/101040
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/101040
2021-12-16 15:39:58.19
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/249782020-11-12T20:37:00Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:23Z
2014-01-23T02:36:23Z
2008
http://hdl.handle.net/10919/24978
http://www.johncairns.net/Commentaries/VT%20Research%20Intro.pdf
Humankind has already passed the first climatic tipping point therefore engaging itself in a global experiment that has already reduced agricultural productivity. With the Artic ice melting 100 years ahead of IPCC prediction, the melting tundra is releasing greenhouse gases that cause temperature increases, leading to more melting tundra. Politicians have set ears, 2025 and 2050, to reduce greenhouse, but Mother Nature is not one to bargain. Once nature s laws are violated the penalties are usually sever and immediate including starvation, disease, and death.
en_US
Virginia Tech Research Magazine
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
tipping points
climate change
global warming
greenhouse gas emissions
Red Alert - Climate Change: This is the Last Chance
Article
Text
ORIGINAL
cairns_red_alert.pdf
application/pdf
99590
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/4b4d2a77-3742-431f-814a-945c5e1eb5ea/download
61111f50215f9c5d2af5d79007ed4941
MD5
1
TEXT
cairns_red_alert.pdf.txt
cairns_red_alert.pdf.txt
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4530
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/808e19ac-009f-4442-a07c-9f681d4f5cdc/download
df0e21814d28f62c7912b1542181ddaa
MD5
2
THUMBNAIL
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cairns_red_alert.pdf.jpg
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ce27d7f99300c95c80e47fc1b67b0bba
MD5
4
10919/24978
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/24978
2020-11-12 20:37:00.666
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/468472022-05-03T13:44:04Zcom_10919_23829com_10919_5553col_10919_23830
Brookshire, E. N. Jack
Valett, H. M.
Thomas, S. A.
Webster, Jackson R.
Biological Sciences
2014-03-27T13:06:06Z
2014-03-27T13:06:06Z
2005-09
2014-03-11
E. N Jack Brookshire, H. Maurice Valett, Steven A. Thomas, and Jackson R. Webster 2005. COUPLED CYCLING OF DISSOLVED ORGANIC NITROGEN AND CARBON IN A FOREST STREAM. Ecology 86:2487-2496. http://dx.doi.org/10.1890/04-1184
0012-9658
http://hdl.handle.net/10919/46847
http://www.esajournals.org/doi/pdf/10.1890/04-1184
https://doi.org/10.1890/04-1184
Dissolved organic nitrogen (DON) is an abundant but poorly understood pool of N in many ecosystems. We assessed DON cycling in a N-limited headwater forest stream via whole-ecosystem additions of dissolved inorganic nitrogen (DIN) and labile dissolved organic matter (DOM), hydrologic transport and biogeochemical modeling, and laboratory experiments with native sediments. We sampled surface and subsurface waters to understand how interaction among hydrologic exchange, DIN, DON, and dissolved organic carbon (DOC) influence stream N losses at summer baseflow. Added DON was taken up rapidly from the water column at rates exceeding DOC and DIN. A significant fraction of this DON was mineralized and nitrified. Combined DON and NO3-N uptake lengths resulted in spiraling lengths of similar to 210 m, suggesting the potential for multiple. transformations of labile N loads within catchment boundaries. Simultaneous addition of DIN increased DOM uptake, but more so for C, resulting in an upward shift in the C:N ratio of uptake. Sediment incubations also showed a strong biotic influence on DOC and DON dynamics. Despite efficient uptake of added DOM, background DON and high molecular mass DOC concentrations increased downstream, resulting in higher DOM loads than could be accounted for by groundwater discharge and suggesting net release of less bioavailable forms from the channel/hyporheic zone. At the same time, subsurface DOM was characterized by very low C:N ratios and a disproportionately large DON pool despite rapid hydrologic mixing with dilute and high C:N ratio surface waters. Analysis of expected DON loads from conservative hyporheic fluxes indicated that watershed losses of DON would have been seven times greater in the absence of apparent benthic demand, suggesting tight internal cycling of subsurface DON. Our study further demonstrates the potential for significant transformation of N in headwater streams before export to downstream ecosystems.
NSF grant DEB 04-05373
en_US
Ecological Society of America
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
cycling
dissolved organic carbon
dissolved organic nitrogen
forest
stream
head-water catchment
hydrologic losses
nutrient uptake
spiraling
transient storage
headwater streams
united-states
dynamics
nutrient
ecosystems
watersheds
retention
patterns
transport
Coupled cycling of dissolved organic nitrogen and carbon in a forest stream
Ecology
Article - Refereed
ORIGINAL
04-1184.pdf
04-1184.pdf
Main article
application/pdf
186317
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/d8f09ad0-8d75-447f-a75e-62d932f95f99/download
d3347f67dec7efd76370b44c5d489478
MD5
1
TEXT
04-1184.pdf.txt
04-1184.pdf.txt
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text/plain
51710
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/ab51b081-2562-439e-869a-5b7be0ea618b/download
22f9c77d52d46d9eecc015ba3b021d40
MD5
2
THUMBNAIL
04-1184.pdf.jpg
04-1184.pdf.jpg
IM Thumbnail
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5200
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/d2b9ba53-d24c-420e-b3de-587b677998ef/download
c09e8dfd946d2e2d37a2511d6234311d
MD5
4
10919/46847
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/46847
2022-05-03 13:44:04.916
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/468322020-11-12T21:00:25Zcom_10919_23829com_10919_5553col_10919_23830
Virginia Tech
Sinsabaugh, R. L.
Linkins, A. E.
Benfield, Ernest F.
Biological Sciences
2014-03-27T13:06:03Z
2014-03-27T13:06:03Z
1985
2014-03-11
R. L. Sinsabaugh, A. E. Linkins, and E. F. Benfield 1985. Cellulose Digestion and Assimilation by Three Leaf-shredding Aquatic Insects. Ecology 66:1464-1471. http://dx.doi.org/10.2307/1938009
0012-9658
http://hdl.handle.net/10919/46832
http://www.jstor.org/stable/1938009
https://doi.org/10.2307/1938009
The capacity of three leaf-shredding aquatic insects, Pteronarcys proteus (Plecoptera: Pteronarcidae), Tipula abdominalis (Diptera: Tipulidae), and Pycnopsyche luculenta (Trichoptera: Limnephilidae), to digest and assimilate cellulose was investigated. Pteronarcys numphs collected from two second-order woodland streams over a 14-mo period exhibited high levels of cellulolytic activity in their alimentary tracts, especially in the anterior gut. Similar though slightly lower activity levels were measured in Pycnopsyche guts. Cellulolytic activity in Tipula larvae collected from the streams during the same period was low to absent, and when present, was concentrated in the hindgut. General proteolytic activity was activity was similar in the alimentary tracts of all three species. Assimilation of uniformly labelled 14C-cellulose was determined by a dual-label technique, and assimilation efficiencies were estimated at 11.2% for Pteronarcys, 18.5% for Tipula, and 12.0% for Pycnopsyche. Confirmation that labelled digestion products passed the gut wall in two species was obtained by in vitro label transport experiments. Ion exchange fractionation of labelled digestion products crossing the gut wall showed >90% of the label was transported as organic acid and amino acids in Tipula, while >40% of the label crossing the gut wall in Pteronarcys was neutral sugar. Based on the label experiments and published information, we hypothesize that Tipula relies mainly on microbial endosymbionts for cellulose hydrolysis, while Pteronarcys accomplishes hydrolysis largely by means of acquired microbial enzymes obtained through ingestion of microbially conditioned detritus. This study demonstrates the potential for certain leaf-shredding stream insect to derive nutritional benefit from plant polysaccharides, although not without microbial mediation.
FMC Corporation through the Virginia State Water Control Board
en_US
Ecological Society of America
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
cellulose
cellulose digestion
Pteronarcys
Pycnopsyche assimilation efficiency
shredder
stream
Tipula
Cellulose digestion and assimilation by three leaf-shredding aquatic insects
Ecology
Article - Refereed
ORIGINAL
1938009.pdf
1938009.pdf
Main article
application/pdf
1596854
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/e46a32f6-448a-4a17-9df1-2c0fa770908b/download
7ef2e551e2e217009dd11ab077d291ee
MD5
1
TEXT
1938009.pdf.txt
1938009.pdf.txt
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text/plain
44775
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/49094b02-4000-4ba3-819f-0a0b2e3c87ef/download
fb26f6fe84c84a17876bb6e924ccb929
MD5
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THUMBNAIL
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1938009.pdf.jpg
IM Thumbnail
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5023
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/df1d1fcc-7050-47f8-9da6-02c5d7f42ac9/download
2f41f84cba10c29764c403f17dee30d7
MD5
4
10919/46832
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/46832
2020-11-12 21:00:25.263
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/953092020-10-22T03:35:28Zcom_10919_23829com_10919_5553col_10919_23830
Lendavi, Ádám
Akçay, Çağlar
Weiss, Talia
Haussmann, Mark F.
Moore, Ignacio T.
Bonier, Frances
Biological Sciences
2019-11-06T18:38:06Z
2019-11-06T18:38:06Z
2015-04-07
http://hdl.handle.net/10919/95309
https://doi.org/10.7717/peerj.877
3
Playbacks of visual or audio stimuli to wild animals is a widely used experimental tool in behavioral ecology. In many cases, however, playback experiments are constrained by observer limitations such as the time observers can be present, or the accuracy of observation. These problems are particularly apparent when playbacks are triggered by specific events, such as performing a specific behavior, or are targeted to specific individuals.We developed a low-cost automated playback/recording system, using two field-deployable devices: radio-frequency identification (RFID) readers and Raspberry Pi micro-computers. This system detects a specific passive integrated transponder (PIT) tag attached to an individual, and subsequently plays back the stimuli, or records audio or visual information. To demonstrate the utility of this system and to test one of its possible applications, we tagged female and male tree swallows (Tachycineta bicolor) from two box-nesting populations with PIT tags and carried out playbacks of nestling begging calls every time focal females entered the nestbox over a six-hour period.We show that the RFID-Raspberry Pi systempresents a versatile, low-cost, field-deployable system that can be adapted for many audio and visual playback purposes. In addition, the set-up does not require programming knowledge, and it easily customized to many other applications, depending on the research questions. Here, we discuss the possible applications and limitations of the system. The low cost and the small learning curve of the RFID-Raspberry Pi system provides a powerful new tool to field biologists.
U.S.National Science Foundation (NSF): IOS-1145625.
Natural Sciences and Engineering Research Council of Canada.
Banting Postdoctoral Fellowship.
Hungarian Scientific Research Fund:OTKA K113108.
11 pages
application/pdf
en
PeerJ
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Animal Behavior
RFID
Automated playback
RPi
Animal communication
Raspberry Pi
Event logging
Parental care
Tree swallow
Low cost audiovisual playback and recording triggered by radio frequency identification using Raspberry Pi
PeerJ
Article - Refereed
Text
ORIGINAL
Low_cost_audiovisual_playback.PDF
Low_cost_audiovisual_playback.PDF
application/pdf
16098685
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/fa9d44bc-e0a1-4914-a8b2-1db98cf905e8/download
2fee02cc7310dcabd39cfff330565e31
MD5
1
TEXT
Low_cost_audiovisual_playback.PDF.txt
Low_cost_audiovisual_playback.PDF.txt
Extracted text
text/plain
30237
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/3964a87c-8bff-423a-a665-11bd6aaf47b9/download
006b23366b31d58f8186dece9b58e7cb
MD5
2
THUMBNAIL
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Low_cost_audiovisual_playback.PDF.jpg
IM Thumbnail
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6253
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/0319897b-993e-4dab-8031-b93636c54571/download
44408dec37577c9e6689e30c9bc739a7
MD5
4
10919/95309
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/95309
2020-10-22 03:35:28.088
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/250902022-03-29T18:37:35Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:32Z
2014-01-23T02:36:32Z
2005
http://hdl.handle.net/10919/25090
http://www.johncairns.net/Commentaries/cairnslifestyle.pdf
Sustainability ethics is the developing effort to ascertain the prerequisites for sustainable use of the planet and also determines the environmental literacy level necessary to persuade people to live sustainably. However, the 21st century lifestyle emphasizes development, which is associated with economic growth, population growth, and increase in material possessions. Currently, these practices are major obstacles to sustainable use of the planet.
en_US
Virginia Tech
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Sustainability
Sustainability ethics
development
global warming
climate change
population growth
Sustainability Ethics and the 2st Century Human Lifestyle
Article
Text
ORIGINAL
cairns_human_lifestyle.pdf
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42090
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2022-03-29 18:37:35.549
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/970912021-10-08T16:43:30Zcom_10919_8195com_10919_25799com_10919_23829com_10919_5553col_10919_78882col_10919_23830
Jenkins, Conor
Orsburn, Benjamin
Biological Sciences
2020-03-02T13:26:08Z
2020-03-02T13:26:08Z
2020-01-31
2020-03-02T12:39:32Z
Jenkins, C.; Orsburn, B. The Cannabis Proteome Draft Map Project. Int. J. Mol. Sci. 2020, 21, 965.
http://hdl.handle.net/10919/97091
https://doi.org/10.3390/ijms21030965
Recently we have seen a relaxation of the historic restrictions on the use and subsequent research on the <i>Cannabis</i> plants, generally classified as <i>Cannabis sativa</i> and <i>Cannabis indica</i>. What research has been performed to date has centered on chemical analysis of plant flower products, namely cannabinoids and various terpenes that directly contribute to phenotypic characteristics of the female flowers. In addition, we have seen many groups recently completing genetic profiles of various plants of commercial value. To date, no comprehensive attempt has been made to profile the proteomes of these plants. We report herein our progress on constructing a comprehensive draft map of the <i>Cannabis</i> proteome. To date we have identified over 17,000 potential protein sequences. Unfortunately, no annotated genome of <i>Cannabis</i> plants currently exists. We present a method by which “next generation” DNA sequencing output and shotgun proteomics data can be combined to produce annotated FASTA files, bypassing the need for annotated genetic information altogether in traditional proteomics workflows. The resulting material represents the first comprehensive annotated protein FASTA for any <i>Cannabis</i> plant. Using this annotated database as reference we can refine our protein identifications, resulting in the confident identification of 13,000 proteins with putative function. Furthermore, we demonstrate that post-translational modifications play an important role in the proteomes of <i>Cannabis</i> flower, particularly lysine acetylation and protein glycosylation. To facilitate the evolution of analytical investigations into these plant materials, we have created a portal to host resources developed from our proteomic and metabolomic analysis of <i>Cannabis</i> plant material as well as our results integrating these resources.
Published version
application/pdf
en
MDPI
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Proteomics
proteogenomics
Cannabis
PTMs
The Cannabis Proteome Draft Map Project
International Journal of Molecular Science
Article - Refereed
Text
ORIGINAL
ijms-21-00965-v2.pdf
application/pdf
3100602
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SWORD
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ijms-21-00965-v2.pdf.txt
ijms-21-00965-v2.pdf.txt
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10919/97091
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/97091
2021-10-08 16:43:30.603
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1159772023-08-04T07:12:08Zcom_10919_23829com_10919_5553col_10919_23830
Rebholz, Zarley
Shewade, Leena
Kaler, Kylie
Larose, Hailey
Schubot, Florian
Tholl, Dorothea
Morozov, Alexandre V.
O'Maille, Paul E.
2023-08-03T14:39:25Z
2023-08-03T14:39:25Z
2023-05
0961-8368
e4634
http://hdl.handle.net/10919/115977
https://doi.org/10.1002/pro.4634
32
5
36974623
1469-896X
Insects have evolved a chemical communication system using terpenoids, a structurally diverse class of specialized metabolites, previously thought to be exclusively produced by plants and microbes. Gene discovery, bioinformatics, and biochemical characterization of multiple insect terpene synthases (TPSs) revealed that isopentenyl diphosphate synthases (IDS), enzymes from primary isoprenoid metabolism, are their likely evolutionary progenitors. However, the mutations underlying the emergence of the TPS function remain a mystery. To address this gap, we present the first structural and mechanistic model for the evolutionary emergence of TPS function in insects. Through identifying key mechanistic differences between IDS and TPS enzymes, we hypothesize that the loss of isopentenyl diphosphate (IPP) binding motifs strongly correlates with the gain of the TPS function. Based on this premise, we have elaborated the first explicit structural definition of isopentenyl diphosphate-binding motifs (IBMs) and used the IBM definitions to examine previously characterized insect IDSs and TPSs and to predict the functions of as yet uncharacterized insect IDSs. Consistent with our hypothesis, we observed a clear pattern of disruptive substitutions to IBMs in characterized insect TPSs. In contrast, insect IDSs maintain essential consensus residues for binding IPP. Extending our analysis, we constructed the most comprehensive phylogeny of insect IDS sequences (430 full length sequences from eight insect orders) and used IBMs to predict the function of TPSs. Based on our analysis, we infer multiple, independent TPS emergence events across the class of insects, paving the way for future gene discovery efforts.
National Science Foundation [MCB1920914, MCB1920922, MCB1920925]
Published version
ACKNOWLEDGMENTS This work was supported by National Science Foundation awards MCB1920914 to Alexandre V. Morozov, MCB1920922 to Paul E. O'Maille, and MCB1920925 to Dorothea Tholl.
application/pdf
en
Wiley
Creative Commons Attribution-NonCommercial 4.0 International
http://creativecommons.org/licenses/by-nc/4.0/
evolution
insects
isoprenoid
isopentenyl diphosphate synthase
mechanism
motifs
phylogeny
structure
terpene
Terpene synthase
Emergence of terpene chemical communication in insects: Evolutionary recruitment of isoprenoid metabolism
Protein Science
Article - Refereed
Text
ORIGINAL
Protein Science - 2023 - Rebholz.pdf
Protein Science - 2023 - Rebholz.pdf
Published version
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oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/115977
2023-08-04 07:12:08.875
http://creativecommons.org/licenses/by-nc/4.0/
Creative Commons Attribution-NonCommercial 4.0 International
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oai:vtechworks.lib.vt.edu:10919/786532023-12-11T11:08:13Zcom_10919_8195com_10919_25799com_10919_11363com_10919_5539com_10919_23829com_10919_5553com_10919_19035com_10919_24210col_10919_18629col_10919_23164col_10919_23830col_10919_24290col_10919_24285
Oguz, Cihan
Watson, Layne T.
Baumann, William T.
Tyson, John J.
Aerospace and Ocean Engineering
Biological Sciences
Computer Science
Mathematics
2017-08-03T20:12:06Z
2017-08-03T20:12:06Z
2017-02-28
2017-08-03T10:59:56Z
BMC Systems Biology. 2017 Feb 28;11(1):30
http://hdl.handle.net/10919/78653
https://doi.org/10.1186/s12918-017-0409-1
Background
Parameter estimation in systems biology is typically done by enforcing experimental observations through an objective function as the parameter space of a model is explored by numerical simulations. Past studies have shown that one usually finds a set of “feasible” parameter vectors that fit the available experimental data equally well, and that these alternative vectors can make different predictions under novel experimental conditions. In this study, we characterize the feasible region of a complex model of the budding yeast cell cycle under a large set of discrete experimental constraints in order to test whether the statistical features of relative protein abundance predictions are influenced by the topology of the cell cycle regulatory network.
Results
Using differential evolution, we generate an ensemble of feasible parameter vectors that reproduce the phenotypes (viable or inviable) of wild-type yeast cells and 110 mutant strains. We use this ensemble to predict the phenotypes of 129 mutant strains for which experimental data is not available. We identify 86 novel mutants that are predicted to be viable and then rank the cell cycle proteins in terms of their contributions to cumulative variability of relative protein abundance predictions. Proteins involved in “regulation of cell size” and “regulation of G1/S transition” contribute most to predictive variability, whereas proteins involved in “positive regulation of transcription involved in exit from mitosis,” “mitotic spindle assembly checkpoint” and “negative regulation of cyclin-dependent protein kinase by cyclin degradation” contribute the least. These results suggest that the statistics of these predictions may be generating patterns specific to individual network modules (START, S/G2/M, and EXIT). To test this hypothesis, we develop random forest models for predicting the network modules of cell cycle regulators using relative abundance statistics as model inputs. Predictive performance is assessed by the areas under receiver operating characteristics curves (AUC). Our models generate an AUC range of 0.83-0.87 as opposed to randomized models with AUC values around 0.50.
Conclusions
By using differential evolution and random forest modeling, we show that the model prediction statistics generate distinct network module-specific patterns within the cell cycle network.
Published version
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
The Author(s)
Predicting network modules of cell cycle regulators using relative protein abundance statistics
BMC Systems Biology
Article - Refereed
Text
ORIGINAL
12918_2017_Article_409.pdf
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oai:vtechworks.lib.vt.edu:10919/78653
2023-12-11 11:08:13.821
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
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oai:vtechworks.lib.vt.edu:10919/1081722022-04-15T20:52:19Zcom_10919_5com_10919_25799com_10919_78629com_10919_78628com_10919_23829com_10919_5553com_10919_86665com_10919_24211com_10919_24214com_10919_5539col_10919_70873col_10919_78630col_10919_23830col_10919_86666col_10919_24288
Behzadinasab, Saeed
Williams, Myra D.
Hosseini, Mohsen
Poon, Leo L. M.
Chin, Alex W. H.
Falkinham, Joseph O. III
Ducker, William A.
2022-02-06T21:57:46Z
2022-02-06T21:57:46Z
2021-11-24
2022-02-06T21:57:36Z
1944-8244
http://hdl.handle.net/10919/108172
https://doi.org/10.1021/acsami.1c15505
13
46
Ducker, William [0000-0002-8207-768X]
34766745
1944-8252
Antimicrobial coatings are one method to reduce the spread of microbial diseases. Transparent coatings preserve the visual properties of surfaces and are strictly necessary for applications such as antimicrobial cell phone screens. This work describes transparent coatings that inactivate microbes within minutes. The coatings are based on a polydopamine (PDA) adhesive, which has the useful property that the monomer can be sprayed, and then the monomer polymerizes in a conformal film at room temperature. Two coatings are described (1) a coating where PDA is deposited first and then a thin layer of copper is grown on the PDA by electroless deposition (PDA/Cu) and (2) a coating where a suspension of Cu2O particles in a PDA solution is deposited in a single step (PDA/Cu2O). In the second coating, PDA menisci bind Cu2O particles to the solid surface. Both coatings are transparent and are highly efficient in inactivating microbes. PDA/Cu kills >99.99% of Pseudomonas aeruginosa and 99.18% of methicillin-resistant Staphylococcus aureus (MRSA) in only 10 min and inactivates 99.98% of SARS-CoV-2 virus in 1 h. PDA/Cu2O kills 99.94% of P. aeruginosa and 96.82% of MRSA within 10 min and inactivates 99.88% of SARS-CoV-2 in 1 h.
Published version
Pages 54706-54714
application/pdf
en
American Chemical Society
https://www.ncbi.nlm.nih.gov/pubmed/34766745
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
COVID-19
SARS-CoV-2 virus
antibacterial
antimicrobial
coating
drug-resistant bacteria
transparent
03 Chemical Sciences
09 Engineering
Nanoscience & Nanotechnology
Humans
Pseudomonas aeruginosa
Anti-Bacterial Agents
Antiviral Agents
Drug Resistance, Microbial
Surface Properties
Methicillin-Resistant Staphylococcus aureus
COVID-19
SARS-CoV-2
Transparent and Sprayable Surface Coatings that Kill Drug-Resistant Bacteria within Minutes and Inactivate SARS-CoV-2 Virus
ACS Applied Materials and Interfaces
Article - Refereed
Journal Article
Text
/Virginia Tech
/Virginia Tech/Engineering
/Virginia Tech/Engineering/Chemical Engineering
/Virginia Tech/All T&R Faculty
/Virginia Tech/Engineering/COE T&R Faculty
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Transparent and Sprayable Surface Coatings that Kill Drug-Resistant Bacteria Within Minutes and Inactivate SARS-CoV-2 Virus.pdf
Transparent and Sprayable Surface Coatings that Kill Drug-Resistant Bacteria Within Minutes and Inactivate SARS-CoV-2 Virus.pdf
Published version
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2022-04-15 20:52:19.15
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1024282021-10-04T14:49:26Zcom_10919_78629com_10919_78628com_10919_23829com_10919_5553col_10919_78630col_10919_23830
Carlson, Colin J.
Hopkins, Skylar R.
Bell, Kayce C.
Dona, Jorge
Godfrey, Stephanie S.
Kwak, Mackenzie L.
Lafferty, Kevin D.
Moir, Melinda L.
Speer, Kelly A.
Strona, Giovanni
Torchin, Mark
Wood, Chelsea L.
Biological Sciences
2021-02-23T14:14:21Z
2021-02-23T14:14:21Z
2020-10
0006-3207
108596
http://hdl.handle.net/10919/102428
https://doi.org/10.1016/j.biocon.2020.108596
250
1873-2917
Found throughout the tree of life and in every ecosystem, parasites are some of the most diverse, ecologically important animals on Earth-but in almost all cases, the least protected by wildlife or ecosystem conservation efforts. For decades, ecologists have been calling for research to understand parasites' important ecological role, and increasingly, to protect as many species from extinction as possible. However, most conservationists still work within priority systems for funding and effort that exclude or ignore parasites, or treat parasites as an obstacle to be overcome. Our working group identified 12 goals for the next decade that could advance parasite biodiversity conservation through an ambitious mix of research, advocacy, and management.
Ecological Society of America; Georgetown Environment Initiative; Smithsonian National Museum of Natural HistorySmithsonian InstitutionSmithsonian National Museum of Natural History; Peter Buck Postdoctoral Fellowship from the Smithsonian National Museum of Natural History; National Science FoundationNational Science Foundation (NSF) [OCE-1829509]; Alfred P. Sloan Foundation Sloan Research FellowshipAlfred P. Sloan Foundation; University of Washington Innovation Award; University of Washington Royalty Research Fund awardUniversity of Washington
The authors kindly thank the Ecological Society of America for supporting our workshop, as well as additional participants Kevin Burgio, Tad Dallas, and Roger Jovani; Laura Whitehouse, for her graphic design work on Fig. 1; Jonathan Wojcik for allowing the inclusion of his copyright Diplozoon illustration in Fig. 3; and dozens of collaborators and friends who have been part of the foundational work on parasite conservation, including Anna Phillips, Veronica Bueno, Carrie Cizauskas, Christopher Clements, Graeme Cumming, Eric Dougherty, Kevin Johnson, Wayne Getz, Nyeema Harris, Elizabeth Nichols, Sergey Mironov, Robert Poulin, and Heather Proctor. CJC gratefully acknowledges funding support from the Georgetown Environment Initiative, and research support from Anna Phillips and the Smithsonian National Museum of Natural History. KCB was supported by a Peter Buck Postdoctoral Fellowship from the Smithsonian National Museum of Natural History. CLW was supported by a grant from the National Science Foundation (OCE-1829509), an Alfred P. Sloan Foundation Sloan Research Fellowship, a University of Washington Innovation Award, and a University of Washington Royalty Research Fund award. Any use of trade, product, or firm names in this publication is for descriptive purposes only and does not imply endorsement by the U.S. Government.
Public domain – authored by a U.S. government employee
application/pdf
en
Public Domain
http://creativecommons.org/publicdomain/mark/1.0/
Biodiversity inventory
Coextinction
Conservation prioritization
Natural history collections
Red listing
Symbionts
A global parasite conservation plan
Biological Conservation
Article - Refereed
Text
StillImage
ORIGINAL
Carlson-et-al.-2020-1.pdf
Carlson-et-al.-2020-1.pdf
application/pdf
3513405
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/04e818ce-50e7-4491-a99b-6e6195139855/download
45c9d4d39a75807f2a316598a7aa0860
MD5
1
TEXT
Carlson-et-al.-2020-1.pdf.txt
Carlson-et-al.-2020-1.pdf.txt
Extracted text
text/plain
94763
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/61968cd2-b2ca-4e51-b729-74e82c4aeb45/download
9a1a2486910641f6f2f471f334aae670
MD5
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THUMBNAIL
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Carlson-et-al.-2020-1.pdf.jpg
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48757f600e4d8f23b8ae24bea8ff3984
MD5
3
10919/102428
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/102428
2021-10-04 14:49:26.832
http://creativecommons.org/publicdomain/mark/1.0/
Public Domain
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1151412023-05-23T07:13:08Zcom_10919_23829com_10919_5553col_10919_23830
Arora, Rajeev
Wisniewski, Michael
Tuong, Tan
Livingston, David
2023-05-22T17:46:35Z
2023-05-22T17:46:35Z
2023-03
0031-9317
http://hdl.handle.net/10919/115141
https://doi.org/10.1111/ppl.13876
175
2
36808742
1399-3054
Evergreen leaves of Rhododendron species inhabiting temperate/montane climates are typically exposed to both high radiation and freezing temperatures during winter when photosynthetic biochemistry is severely inhibited. Cold-induced "thermonasty, " that is, lamina rolling and petiole curling, can reduce the amount of leaf area exposed to solar radiation and has been associated with photoprotection in overwintering rhododendrons. The present study was conducted on natural, mature plantings of a cold-hardy and large-leaved thermonastic North American species (Rhododendron maximum) during winter freezes. Infrared thermography was used to determine initial sites of ice formation, patterns of ice propagation, and dynamics of the freezing process in leaves to understand the temporal and mechanistic relationship between freezing and thermonasty. Results indicated that ice formation in whole plants is initiated in the stem, predominantly in the upper portions, and propagates in both directions from the original site. Ice formation in leaves initially occurred in the vascular tissue of the midrib and then propagated into other portions of the vascular system/venation. Ice was never observed to initiate or propagate into palisade, spongy mesophyll, or epidermal tissues. These observations, together with the leaf- and petiole-histology, and a simulation of the rolling effect of dehydrated leaves using a cellulose-based, paper-bilayer system, suggest that thermonasty occurs due to anisotropic contraction of cell wall cellulose fibers of adaxial versus abaxial surface as the cells lose water to ice present in vascular tissues.
Iowa Agriculture and Home Economics Experiment Station, Hatch Act and State of Iowa [3601]
Published version
Iowa Agriculture and Home Economics Experiment Station, Hatch Act and State of Iowa, Grant/Award Number: 3601
Public domain – authored by a U.S. government employee
application/pdf
en
Wiley
Public Domain (U.S.)
http://creativecommons.org/publicdomain/mark/1.0/
Leaf movements
ice nucleation
propagation
cold
temperature
plants
water
catawbiense
expression
pattern
Infrared thermography of in situ natural freezing and mechanism of winter-thermonasty in Rhododendron maximum
Physiologia Plantarum
Article - Refereed
Text
ORIGINAL
AroraInfrared.pdf
AroraInfrared.pdf
Published version
application/pdf
18448739
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/b457a7a9-fde7-4cf1-9c4b-97b89fbb7eee/download
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MD5
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TEXT
AroraInfrared.pdf.txt
AroraInfrared.pdf.txt
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text/plain
56791
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/8ed4e4f7-c264-4ac4-bfa4-1ff8fbae91b2/download
764e96e8be0564e9b320c72d731ce70d
MD5
2
THUMBNAIL
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AroraInfrared.pdf.jpg
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https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/267789c6-88c3-4c0e-9b25-433165a452ac/download
c04086192a7e0b67eeff37b29352082f
MD5
3
10919/115141
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/115141
2023-05-23 07:13:08.732
http://creativecommons.org/publicdomain/mark/1.0/
Public Domain (U.S.)
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/250752022-03-29T18:37:36Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:31Z
2014-01-23T02:36:31Z
2005
http://hdl.handle.net/10919/25075
http://www.johncairns.net/Commentaries/TowerofBabel.pdf
The key to long-term economic and social stability is the sustainable use of the planet. The Global Tower of Babel is such a formidable obstacle in achieving sustainable use of the planet that the barriers must be diminished. However, if a global consensus can be reached and compatible eco-ethics and sustainability ethics can be established, humankind may have a chance at achieving sustainability despite its cultural diversity.
en_US
Virginia Tech
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Sustainability
eco-ethics
cultural diversity
resource wars
The Global Tower of Babel
Article
Text
ORIGINAL
cairns_global_tower_babel.pdf
application/pdf
24657
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/a4f945e8-954a-4e8d-b3c2-feec62fe89cf/download
37bfdd244a7bc2a988253d88fe9ef41f
MD5
1
TEXT
cairns_global_tower_babel.pdf.txt
cairns_global_tower_babel.pdf.txt
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text/plain
9376
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/1a062379-c40f-4a97-a718-c8b036cdf895/download
9b44fca8c583fc6a46b4bf818e4c2eed
MD5
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THUMBNAIL
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https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/b7fc1d58-65d8-4892-8e17-1b456185e2c4/download
911b0f908bacd820cf559b60e8bc58a0
MD5
4
10919/25075
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/25075
2022-03-29 18:37:36.067
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/766312023-11-29T19:07:45Zcom_10919_5540com_10919_23829com_10919_5553com_10919_91436com_10919_5532col_10919_71752col_10919_23830col_10919_97229
Redekar, Neelam R.
Clevinger, Elizabeth M.
Laskar, M. A.
Biyashev, Ruslan M.
Ashfield, Tom
Jensen, Roderick V.
Jeong, Soon-Chun
Tolin, Sue A.
Saghai-Maroof, Mohammad A.
Biological Sciences
School of Plant and Environmental Sciences
2017-03-13T17:13:40Z
2017-03-13T17:13:40Z
2016-05-13
http://hdl.handle.net/10919/76631
https://doi.org/10.3835/plantgenome2015.09.0088
9
2
Rsv3 is one of three genetic loci conferring strain-specific resistance to Soybean mosaic virus (SMV). The Rsv3 locus has been mapped to a 154-kb region on chromosome 14, containing a cluster of five nucleotide-binding leucine-rich repeat (NB-LRR) resistance genes. High sequence similarity between the Rsv3 candidate genes challenges fine mapping of the locus. Among the five, Glyma14g38533 showed the highest transcript abundance in 1 to 3 h of SMV-G7 inoculation. Comparative sequence analyses were conducted with the five Rsv3 candidate NB-LRR genes from susceptible (rsv-type) soybean [Glycine max (L.) Merr.] cultivar Williams 82, resistant (Rsv3-type) cultivar Hwangkeum, and resistant lines L29 and RRR. Sequence comparisons revealed that Glyma14g38533 had far more polymorphisms than the other candidate genes. Interestingly, Glyma14g38533 gene from Rsv3-type lines exhibited 150 single-nucleotide polymorphism (SNP and six insertion–deletion (InDel) markers relative to rsv-type line, Furthermore, the polymorphisms identified in three Rsv3-type lines were highly conserved. Several polymorphisms were validated in 18 Rsv3-type resistant and six rsv-type susceptible lines and were found associated with their disease response. The majority of the polymorphisms were located in LRR domain encoding region, which is involved in pathogen recognition via protein–protein interactions. These findings associating Glyma14g38533 with Rsv3-type resistance to SMV suggest it is the most likely candidate gene for Rsv3.
application/pdf
en
Crop Science Society of America
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
Candidate Gene Sequence Analyses toward Identifying Rsv3-Type Resistance to Soybean Mosaic Virus
The Plant Genome
Article - Refereed
Text
ORIGINAL
RedekarCandidate2016.pdf
RedekarCandidate2016.pdf
application/pdf
1702299
https://vtechworks.lib.vt.edu/bitstreams/66354c07-1f06-497e-823c-db2fdac0ee97/download
fc4f972ac24354ab9042082f1c5e6953
MD5
1
TEXT
RedekarCandidate2016.pdf.txt
RedekarCandidate2016.pdf.txt
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text/plain
57654
https://vtechworks.lib.vt.edu/bitstreams/fe6228f9-f118-4ce8-99ed-d5583ce55d2b/download
16ecc6cb422eba714460c1c8487b52f4
MD5
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THUMBNAIL
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b20e7d6b38f3d04a175cb998e6232bea
MD5
8
10919/76631
oai:vtechworks.lib.vt.edu:10919/76631
2023-11-29 19:07:45.99
http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/249992022-03-29T18:37:35Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:25Z
2014-01-23T02:36:25Z
2010
http://hdl.handle.net/10919/24999
http://www.johncairns.net/Commentaries/incompatible.pdf
The quest for sustainable use of the planet is admirable, however the flagrant waste of biospheric resources overrides any efforts towards this goal. Although there is not single solution to achieve immediate sustainability, ending the waste of biospheric resources, especially food and energy resources, would be a step in the right direction.
en_US
Virginia Tech
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Sustainability
food resources
energy resources
exploitation
climate change
Sustainability and Waste of Biospheric Resources are Incompatible
Article
Text
ORIGINAL
cairns_biospheric_resources.pdf
application/pdf
15708
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/a4403d73-a79b-4617-a644-3557877b36bb/download
84791da349c5ac75d408d699049041fe
MD5
1
TEXT
cairns_biospheric_resources.pdf.txt
cairns_biospheric_resources.pdf.txt
Extracted text
text/plain
5906
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/f620a956-1a74-491b-af6f-e358bc3328a4/download
061f08497f14d6eb23d9de1db535db71
MD5
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cairns_biospheric_resources.pdf.jpg
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c88e77c8417226d94f06d042d141f23f
MD5
4
10919/24999
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/24999
2022-03-29 18:37:35.761
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/250892020-11-12T20:37:01Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:32Z
2014-01-23T02:36:32Z
2011
http://hdl.handle.net/10919/25089
http://www.johncairns.net/Papers/humaneffects.ppt
Persuasive evidence indicates that the biosphere is experiencing a major biotic crisis and even if humankind ceases stress on natural systems, the crisis will most likely disrupt or alter the surviving ecosystems. The new altered biosphere will be difficult to understand and adapt to within the next five to ten generations. Extinction is a continual process; however, at great intervals, a mass extinction occurs and new species will replace most of the extinct species. The global problems caused by altered climate and other man made problems can only be successfully solved by a civil discourse and a commitment to a common goal by all, or a large majority of, the world s sovereign nations. The only hope for humankind to resolve the dangerous global crises it now faces is through rapid social evolution. It is possible to delay the onset of a major extinction of species by nurturing and protecting the present biosphere. If the present biosphere is not saved, the new altered biosphere may or may not be a suitable habitat for Homo sapiens.
en_US
National Academy of Sciences
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
climate change
global warming
extinction
habitats
evolution
Human Effects Upon Revolutionary Processes in the Biosphere
Supercourse Lectures
Presentation
Text
ORIGINAL
cairns_human_effects.ppt
application/vnd.ms-powerpoint
203264
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/b9223de0-025e-43dc-b5e6-6d453c0c6931/download
3786287e1a95f6ba839b7da55cc4934a
MD5
1
TEXT
cairns_human_effects.ppt.txt
cairns_human_effects.ppt.txt
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text/plain
8758
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/55ae70cb-14ee-4b46-a993-4516843b7bc4/download
78d3d258c3a174b70263156c497aa290
MD5
2
10919/25089
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/25089
2020-11-12 20:37:01.29
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1023272022-02-26T22:17:38Zcom_10919_23829com_10919_5553col_10919_23830
Hamdan, Mohammed
Byström, Pär
Hotchkiss, Erin R.
Al-Haidarey, Mohammed J.
Karlsson, Jan
Biological Sciences
2021-02-10T17:21:29Z
2021-02-10T17:21:29Z
2021-01-08
0046-5070
http://hdl.handle.net/10919/102327
https://doi.org/10.1111/fwb.13679
1365-2427
Climate changes are predicted to influence gross primary production (GPP) of lakes directly through warming and indirectly through increased loads of allochthonous coloured dissolved organic matter (cDOM) from surrounding landscapes. However, few studies have investigated this combined effect. Here we tested the effects of warming (elevated 3celcius) and cDOM input (three levels of humic river water addition) on GPP in autumn (2 months including open water and ice-covered periods) in experimental pond ecosystems. The cDOM input decreased whole-ecosystem GPP at natural temperature conditions mainly as a result of lower benthic GPP not fully counteracted by an increase in pelagic GPP, while warming increased whole-ecosystem GPP due to a positive response of mainly pelagic GPP at all levels of cDOM input. Warming delayed autumn ice cover formation by 2 weeks but did not affect light availability in the water column compared to ambient ice-covered treatments. Gross primary production during this period was still affected by warming and cDOM. The results stress the importance of accounting for multiple climate drivers and habitats when predicting lake GPP responses to climate change. We conclude that climate change may shift whole-ecosystem GPP through different responses of habitat-specific GPP to increasing cDOM inputs and warming.
Ministry of Higher Education and Scientific Research in Iraq; Kunt and Alice Wallenberg FoundationKnut & Alice Wallenberg Foundation
The Ministry of Higher Education and Scientific Research in Iraq; The Kunt and Alice Wallenberg Foundation.
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
brownification
habitat-specific
limitation
rising temperature
whole productivity
An experimental test of climate change effects in northern lakes: Increasing allochthonous organic matter and warming alters autumn primary production
Freshwater Biology
Article - Refereed
Text
StillImage
ORIGINAL
fwb.13679.pdf
fwb.13679.pdf
application/pdf
1272399
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/caecdf05-117f-48bc-8f85-9981ae4a9b9c/download
26f39315d87770c80a0e9717675f72b6
MD5
1
TEXT
fwb.13679.pdf.txt
fwb.13679.pdf.txt
Extracted text
text/plain
53270
https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/c8ab48a2-4709-44f3-b09a-34a5feae5020/download
99bdf289ae9a032592bf631116a25929
MD5
2
THUMBNAIL
fwb.13679.pdf.jpg
fwb.13679.pdf.jpg
IM Thumbnail
image/jpeg
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https://vtw-prod.eed0.cloud.lib.vt.edu/bitstreams/e835c7d1-a497-45c4-a54b-118e73e466bc/download
a13c607dfb32fff402ed7a3a9d72ced9
MD5
3
10919/102327
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/102327
2022-02-26 22:17:38.121
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1175692024-03-15T14:49:15Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553col_10919_70873col_10919_23830
Truchon, A. R.
Chase, E. E.
Gann, E. R.
Moniruzzaman, M.
Creasey, B. A.
Aylward, Frank O.
Xiao, C.
Gobler, C. J.
Wilhelm, S. W.
2024-01-22T18:42:43Z
2024-01-22T18:42:43Z
2023-11-30
1664-302X
https://hdl.handle.net/10919/117569
https://doi.org/10.3389/fmicb.2023.1284617
14
Aylward, Frank [0000-0002-1279-4050]
38098665
1664-302X
Since the discovery of the first “giant virus,” particular attention has been paid toward isolating and culturing these large DNA viruses through Acanthamoeba spp. bait systems. While this method has allowed for the discovery of plenty novel viruses in the Nucleocytoviricota, environmental -omics-based analyses have shown that there is a wealth of diversity among this phylum, particularly in marine datasets. The prevalence of these viruses in metatranscriptomes points toward their ecological importance in nutrient turnover in our oceans and as such, in depth study into non-amoebal Nucleocytoviricota should be considered a focal point in viral ecology. In this review, we report on Kratosvirus quantuckense (née Aureococcus anophagefferens Virus), an algae-infecting virus of the Imitervirales. Current systems for study in the Nucleocytoviricota differ significantly from this virus and its relatives, and a litany of trade-offs within physiology, coding potential, and ecology compared to these other viruses reveal the importance of K. quantuckense. Herein, we review the research that has been performed on this virus as well as its potential as a model system for algal-virus interactions.
Published version
application/pdf
en
Frontiers
https://www.ncbi.nlm.nih.gov/pubmed/38098665
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Aureococcus anophagefferens
Nucleocytoviricota
brown tide
harmful algal blooms
marine microbiology
model system
strain heterogeneity
viral ecology
Kratosvirus quantuckense: the history and novelty of an algal bloom disrupting virus and a model for giant virus research
Frontiers in Microbiology
Article - Refereed
Text
2023-10-30
/Virginia Tech
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/All T&R Faculty
/Virginia Tech/Science/COS T&R Faculty
ORIGINAL
fmicb-14-1284617.pdf
Published version
application/pdf
12857186
https://vtechworks.lib.vt.edu/bitstreams/78be8f74-3a9f-467d-8950-715b59db4fb2/download
24457a11e0d1566aae127a1dd27ce0ef
MD5
1
LICENSE
license.txt
text/plain
1534
https://vtechworks.lib.vt.edu/bitstreams/e6c3ff95-aaab-40c2-a828-92fa936744dc/download
94f9fe93d7cf5c8ee48bc758ea4fa773
MD5
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TEXT
fmicb-14-1284617.pdf.txt
fmicb-14-1284617.pdf.txt
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text/plain
100304
https://vtechworks.lib.vt.edu/bitstreams/078bd0ae-4d5a-470a-be22-b4f95a52e435/download
bb894579e900640a50b55fa29c471e0e
MD5
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THUMBNAIL
fmicb-14-1284617.pdf.jpg
fmicb-14-1284617.pdf.jpg
IM Thumbnail
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https://vtechworks.lib.vt.edu/bitstreams/3b70b080-9ae4-4b80-85cd-599c5ba64e6b/download
9a7c98e088a81403f7d21bfcb136cb5f
MD5
4
10919/117569
oai:vtechworks.lib.vt.edu:10919/117569
2024-03-15 14:49:15.969
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
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oai:vtechworks.lib.vt.edu:10919/248022022-03-21T08:06:44Zcom_10919_23829com_10919_5553col_10919_23830
Virginia Tech
Irwin, Rebecca E.
Adler, Lynn S.
Brody, Alison K.
Biological Sciences
2014-01-10T20:07:55Z
2014-01-10T20:07:55Z
2004-06
2014-01-08
Rebecca E. Irwin, Lynn S. Adler, and Alison K. Brody 2004. THE DUAL ROLE OF FLORAL TRAITS: POLLINATOR ATTRACTION AND PLANT DEFENSE. Ecology 85:1503–1511. http://dx.doi.org/10.1890/03-0390
0012-9658
http://hdl.handle.net/10919/24802
http://www.esajournals.org/doi/pdf/10.1890/03-0390
https://doi.org/10.1890/03-0390
Plants are under siege from a diversity of enemies that consume both leaf and floral parts. Plants resist damage to leaves in a variety of ways, and we now have a rich literature documenting how plants defend themselves against herbivore attack. In contrast, the mechanisms by which plants resist enemies that consume floral parts or resources are much less known, even though damage to floral tissue usually has tighter links to plant fitness than damage to leaf tissue. Many plants experience nectar robbing, whereby floral visitors remove nectar from flowers, often without pollinating. Nectar robbers can reduce plant fitness to degrees comparable to, or even surpassing, reduction by herbivores. However, because nectar attracts both pollinators and nectar robbers, plants face a dilemma in defending against nectar robbers without also deterring pollinators. Here, we extend the conceptual framework of resistance to herbivores to include resistance to nectar robbers, focusing on nectar traits. We review published data and find that an array of nectar traits may deter robbers without deterring pollinators. Although resistance traits against robbers have been broadly identified, the costs and benefits of these traits in terms of plant fitness remain poorly understood. We present data showing that a nectar trait (dilute nectar) might directly, as well as indirectly, benefit plant fitness by deterring nectar-robbing bumble bees of Ipomopsis aggregata without deterring hummingbird pollinators. However, the magnitude of any plant fitness benefit will depend on the degree to which plants are pollen- vs. resource-limited in a given year. The results of our work offer both conceptual and empirical insight into how plants cope with attack by nonpollinating floral visitors through a relatively unexplored trait, nectar.
en_US
Ecological Society of America
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
Bombus occidentalis
humble bee
herbivory
Ipomopsis aggregata
nectar
concentration
nectar robbing
plant dejense
pollination
resistance
ipomopsis-aggregata polemoniaceae
induced resistance
catalpa-speciosa
nectar thieves
scarlet-gilia
toxic nectar
herbivory
fitness
hummingbird
evolution
Environmental Sciences & Ecology
The dual role of floral traits: Pollinator attraction and plant defense
Ecology
Article - Refereed
ORIGINAL
03-0390.pdf
03-0390.pdf
Main article
application/pdf
136105
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03-0390.pdf.txt
03-0390.pdf.txt
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MD5
4
10919/24802
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/24802
2022-03-21 08:06:44.963
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1127032023-11-29T11:25:01Zcom_10919_8195com_10919_25799com_10919_23829com_10919_5553com_10919_24259com_10919_5559com_10919_91913com_10919_23198com_10919_25796com_10919_91912col_10919_18629col_10919_23830col_10919_24342col_10919_91914col_10919_25797col_10919_91916
Xu, Xiguang
Johnson, Zachary
Wang, Amanda
Padget, Rachel L.
Smyth, James W.
Xie, Hehuang
2022-11-28T13:46:23Z
2022-11-28T13:46:23Z
2022-11-23
2022-11-27T04:12:10Z
BMC Biology. 2022 Nov 23;20(1):261
http://hdl.handle.net/10919/112703
https://doi.org/10.1186/s12915-022-01467-0
Background
Folate is an essential B-group vitamin and a key methyl donor with important biological functions including DNA methylation regulation. Normal neurodevelopment and physiology are sensitive to the cellular folate levels. Either deficiency or excess of folate may lead to neurological disorders. Recently, folate has been linked to tRNA cytosine-5 methylation (m5C) and translation in mammalian mitochondria. However, the influence of folate intake on neuronal mRNA m5C modification and translation remains largely unknown. Here, we provide transcriptome-wide landscapes of m5C modification in poly(A)-enriched RNAs together with mRNA transcription and translation profiles for mouse neural stem cells (NSCs) cultured in three different concentrations of folate.
Results
NSCs cultured in three different concentrations of folate showed distinct mRNA methylation profiles. Despite uncovering only a few differentially expressed genes, hundreds of differentially translated genes were identified in NSCs with folate deficiency or supplementation. The differentially translated genes induced by low folate are associated with cytoplasmic translation and mitochondrial function, while the differentially translated genes induced by high folate are associated with increased neural stem cell proliferation. Interestingly, compared to total mRNAs, polysome mRNAs contained high levels of m5C. Furthermore, an integrative analysis indicated a transcript-specific relationship between RNA m5C methylation and mRNA translation efficiency.
Conclusions
Altogether, our study reports a transcriptome-wide influence of folate on mRNA m5C methylation and translation in NSCs and reveals a potential link between mRNA m5C methylation and mRNA translation.
Published version
application/pdf
en
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
The Author(s)
Folate regulates RNA m5C modification and translation in neural stem cells
BMC Biology
Article - Refereed
Text
ORIGINAL
12915_2022_Article_1467.pdf
Published version
application/pdf
3806029
https://vtechworks.lib.vt.edu/bitstreams/c7bee903-da28-4f3d-8b76-59af790533e7/download
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MD5
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LICENSE
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12915_2022_Article_1467.pdf.txt
12915_2022_Article_1467.pdf.txt
Extracted text
text/plain
71381
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10919/112703
oai:vtechworks.lib.vt.edu:10919/112703
2023-11-29 11:25:01.851
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/468402022-03-21T08:03:45Zcom_10919_23829com_10919_5553col_10919_23830
Virginia Tech
Adler, Lynn S.
Bronstein, Judith L.
Biological Sciences
2014-03-27T13:06:05Z
2014-03-27T13:06:05Z
2004-06
2014-03-11
Lynn S. Adler and Judith L. Bronstein 2004. ATTRACTING ANTAGONISTS: DOES FLORAL NECTAR INCREASE LEAF HERBIVORY? Ecology 85:1519-1526. http://dx.doi.org/10.1890/03-0409
0012-9658
http://hdl.handle.net/10919/46840
http://www.esajournals.org/doi/pdf/10.1890/03-0409
https://doi.org/10.1890/03-0409
Traits that are attractive to mutualists may also attract antagonists, resulting in conflicting selection pressures. Here we develop the idea that increased floral nectar production can, in some cases, increase herbivory. In these situations, selection for increased nectar production to attract pollinators may be constrained by a linked cost of herbivore attraction. In support of this hypothesis, we report that experimentally supplementing nectar rewards in Datura stramonium led to increased oviposition by Manduca sexta, a sphingid moth that pollinates flowers, but whose larvae feed on leaf tissue. We speculate that nectar composition may provide information about plant nutritional status or defense that floral visitors could use to make oviposition decisions. Thus, selection by floral visitors and leaf herbivores may be inextricably intertwined, and herbivores may represent a relatively unexplored agent of selection on nectar traits.
Virginia Tech Department of Biology
NSF DEB-0211480
en_US
Ecological Society of America
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
datura stramonium
herbivory
manduca sexta
mutualism
natural
selection
nectar composition
nectar reward
oviposition
pollination
flower-color polymorphism
optimal defense theory
datura-stramonium
wild radish
plant fitness
evolutionary ecology
natural-selection
ipomopsis-aggregata
ipomoea purpurea
seed predators
Attracting antagonists: does floral nectar increase leaf herbivory?
Ecology
Article - Refereed
ORIGINAL
03-0409.pdf
03-0409.pdf
Main article
application/pdf
226914
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03-0409.pdf.txt
03-0409.pdf.txt
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10919/46840
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/46840
2022-03-21 08:03:45.049
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1179062024-03-15T15:09:11Zcom_10919_5com_10919_25799com_10919_104237com_10919_5553com_10919_23829com_10919_25796com_10919_91912com_10919_23198col_10919_70873col_10919_104238col_10919_23830col_10919_25797col_10919_91916
Ahuja, Shreya
Lazar, Iulia M.
2024-02-09T13:24:57Z
2024-02-09T13:24:57Z
2023-07-20
109433 (Article number)
2352-3409
PMC10404857
S2352-3409(23)00533-4 (PII)
https://hdl.handle.net/10919/117906
https://doi.org/10.1016/j.dib.2023.109433
49
Lazar, Maria [0000-0001-7746-7889]
37554993
2352-3409
The immunoprotective functions of microglia in the brain are mediated by the inflammatory M1 phenotype. This phenotype is challenged by anti-inflammatory cytokines which polarize the microglia cells to an immunosuppressive M2 phenotype, a trait that is often exploited by cancer cells to evade immune recognition and promote tumor growth. Investigating the molecular determinants of this behavior is crucial for advancing the understanding of the mechanisms that cancer cells use to escape immune attack. In this article, we describe liquid chromatography (LC)-mass spectrometry (MS)/proteomic data acquired with an EASY-nanoLC 1200-Q ExactiveTM OrbitrapTM mass spectrometer that reflect the response of human microglia cells (HMC3) to stimulation with potential cancer-released anti-inflammatory cytokines known to be key players in promoting tumorigenesis in the brain (IL-4, IL-13, IL-10, TGFB and MCP-1). The MS files were processed with the Proteome Discoverer v.2.4 software package. The cell culture conditions, the sample preparation protocols, the MS acquisition parameters, and the data processing approach are described in detail. The RAW and processed MS files associated with this work were deposited in the PRIDE partner repository of the ProteomeXchange Consortium with the dataset identifiers PXD023163 and PXD023166, and the analyzed data in the Mendeley Data cloud-based repository with DOI 10.17632/fvhw2zwt5d.1. The biological interpretation of the data can be accessed in the research article “Systems-Level Proteomics Evaluation of Microglia Response to Tumor-Supportive Anti-inflammatory Cytokines” (Shreya Ahuja and Iulia M. Lazar, Frontiers in Immunology 2021 [1]). The proteome data described in this article will benefit researchers who are either interested in re-processing the data with alternative search engines and filtering criteria, and/or exploring the data in more depth to advance the understanding of cancer progression and the discovery of novel biomarkers or drug targets.
Published version
application/pdf
en
Elsevier
https://www.ncbi.nlm.nih.gov/pubmed/37554993
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
Anti-inflammatory cytokines
Cancer
Microglia
Proteomics
Anti-inflammatory cytokine stimulation of HMC3 cells: Proteome dataset
Data in Brief
Article - Refereed
Journal
Text
2023-07-17
/Virginia Tech
/Virginia Tech/Science
/Virginia Tech/Science/Biological Sciences
/Virginia Tech/Faculty of Health Sciences
/Virginia Tech/All T&R Faculty
/Virginia Tech/Science/COS T&R Faculty
/Virginia Tech/VT Carilion School of Medicine
/Virginia Tech/VT Carilion School of Medicine/Surgery
/Virginia Tech/VT Carilion School of Medicine/Psychiatry and Behavioral Medicine
/Virginia Tech/VT Carilion School of Medicine/Psychiatry and Behavioral Medicine/Secondary Appointment-Psychiatry and Behavioral Medicine
/Virginia Tech/VT Carilion School of Medicine/Surgery/Secondary Appointment-Surgery
ORIGINAL
Anti-inflammatory cytokine stimulation of HMC3 cells Proteome dataset.pdf
Published version
application/pdf
1494719
https://vtechworks.lib.vt.edu/bitstreams/794fd945-2ad8-4a46-bea6-c1ebb261d19f/download
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MD5
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Anti-inflammatory cytokine stimulation of HMC3 cells Proteome dataset.pdf.txt
Anti-inflammatory cytokine stimulation of HMC3 cells Proteome dataset.pdf.txt
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10919/117906
oai:vtechworks.lib.vt.edu:10919/117906
2024-03-15 15:09:11.43
http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
open.access
https://vtechworks.lib.vt.edu
VTechWorks
vtechworks@vt.edu
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oai:vtechworks.lib.vt.edu:10919/249762020-11-12T20:37:03Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:22Z
2014-01-23T02:36:22Z
2007
http://hdl.handle.net/10919/24976
http://www.johncairns.net/Papers/1._Preparing_to_Monitor.pdf
Monitoring humankind's activities is common, for example, intensive care patients in hospitals, pharmaceuticals, food supplies, industrial processes, and economic health. Basically, monitoring is used to confirm that previously established quality control conditions are being met. If they are not, remedial measures are taken promptly. Monitoring for sustainable use of the planet will be orders of magnitude more complex than the systems just mentioned, but the principle remains the same - a feedback loop providing information about the system of concern is essential to verify that the system is functioning within acceptable limits. The absence of a sound monitoring system significantly increases the probability of unpleasant surprises. Since humankind must live sustainably for at least several generations to increase confidence that its practices are sustainable, the development time for the monitoring system will be extensive. Choosing the attributes to be monitored will be challenging, and the synthesis and integration of such diverse information will require much skill, especially in communication. However, the computer age makes coping with vast amounts of diverse information possible for the first time in human history. Paying attention to seemingly inconsequential components of sustainability, such as individual commitment, has merit.
en_US
Asian Journal of Experimental Sciences
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
sustainaility monitoring
resource wars
environmental quality control
information feedback loops
biospheric life support system
Preparing to Monitor for Sustainable Use of the Planet
Article
Text
ORIGINAL
cairns_preparing_to_monitor.pdf
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168276
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cairns_preparing_to_monitor.pdf.txt
cairns_preparing_to_monitor.pdf.txt
Extracted text
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38491
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10919/24976
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/24976
2020-11-12 20:37:03.118
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1019472023-05-18T13:28:34Zcom_10919_5com_10919_25799com_10919_23829com_10919_5553com_10919_24227com_10919_5532com_10919_91436col_10919_70873col_10919_23830col_10919_24302col_10919_97229
Lakoba, Vasiliy T.
Wind, Lauren L.
DeVilbiss, Stephen
Lofton, Mary E.
Bretz, Kristen
Weinheimer, Alaina R.
Moore, Chloe
Baciocco, Colin
Hotchkiss, Erin R.
Hession, W. Cully
Biological Systems Engineering
School of Plant and Environmental Sciences
Biological Sciences
2021-01-18T19:53:52Z
2021-01-18T19:53:52Z
2020-11-09
2021-01-18T19:53:49Z
http://hdl.handle.net/10919/101947
https://doi.org/10.1021/acsestwater.0c00160
Hession, William [0000-0002-6323-3827]
Anthropogenic freshwater salinization is increasing with global change. Rising freshwater salinity threatens ecosystem biodiversity, health, and services via toxicity to organisms and mobilization of nutrients and metals. Brining roads is one major source of freshwater salinization that continues to grow with rising urbanization. While the detrimental effects of salinization in streams are well-documented, high-frequency, temporal patterns in salt transport, particularly during winter road de-icing in mixed land use landscapes, are less understood. To address this knowledge gap, we analyzed high-frequency specific conductance as a proxy for salinity across 114 high-flow events from 2013 to 2018 in an impaired stream draining mixed agriculture−urban land use. The specific conductance was highest in winter (median = 947 μS cm−1) and decreased with first-order kinetics up to 90 days after brining (β1 = −0.003), suggesting lasting impacts of road de-icing on water quality. Although hysteresis patterns suggested a transition from distal to proximal salt sources, they showed no clear correlation of flushing versus dilution to brining events. While seasonal brining increased salinity in receiving streams, unpredictable transport dynamics reduced the efficacy of hysteresis in characterizing salt transport dynamics. Thus, the complexity of mixed land use watersheds requires more spatially and temporally explicit monitoring to characterize stream salinization dynamics.
Published (Publication status)
application/pdf
en
In Copyright (InC) - Non-Commercial Use Permitted
http://rightsstatements.org/vocab/InC-NC/1.0/
salt
freshwater
salinization
road salts
stream ecology
land use
urbanization
Stroubles Creek
Salt dilution and flushing dynamics of an impaired agricultural−urban stream
ES&T: Water
Article - Refereed
Article
Text
/Virginia Tech/Agriculture & Life Sciences
/Virginia Tech/All T&R Faculty
/Virginia Tech/Agriculture & Life Sciences/Biological Systems Engineering
/Virginia Tech/Agriculture & Life Sciences/CALS T&R Faculty
/Virginia Tech
TEXT
Lakoba2020_acsestwater.0c00160.pdf.txt
Lakoba2020_acsestwater.0c00160.pdf.txt
Extracted text
text/plain
53422
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Original SWORD entry document
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Lakoba2020_acsestwater.0c00160.pdf
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oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/101947
2023-05-18 13:28:34.041
http://rightsstatements.org/vocab/InC-NC/1.0/
In Copyright (InC) - Non-Commercial Use Permitted
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/468142023-11-17T20:34:04Zcom_10919_23829com_10919_5553col_10919_23830
Eggert, Susan L.
Wallace, J. Bruce
Meyer, J. L.
Webster, Jackson R.
Biological Sciences
2014-03-27T13:05:58Z
2014-03-27T13:05:58Z
2012-09
2014-03-11
Sue L. Eggert, J. Bruce Wallace, Judy L. Meyer, and Jackson R. Webster 2012. Storage and export of organic matter in a headwater stream: responses to long-term detrital manipulations. Ecosphere 3:art75. http://dx.doi.org/10.1890/ES12-00061.1
2150-8925
http://hdl.handle.net/10919/46814
http://www.esajournals.org/doi/pdf/10.1890/ES12-00061.1
https://doi.org/10.1890/es12-00061.1
Riparian habitats provide organic matter inputs that influence stream biota and ecosystem processes in forested watersheds. Over a 13-yr period, we examined the effects of litter exclusion, small- and large-wood removal, and the addition of leaf species of varying detrital quality on organic matter standing crop and export of organic and inorganic particles in a high-gradient headwater stream. Using eight pretreatment years of export data and two pretreatment years of particulate organic matter (POM) standing crop data, we report on 21 and 15 years of continuous export and POM standing crop results, respectively. Litter exclusion resulted in the elimination of leaf standing crop by the end of year three. Wood and fine benthic organic matter (FBOM) standing crops declined significantly during the exclusion and wood removal periods, but never completely disappeared. Following the introduction of artificial wood structures for retention, the addition of fast, slow, and mixed breakdown leaves in the treatment stream resulted in significantly increased mean annual leaf standing crops. After five years of leaf addition, FBOM standing crop and fine particulate organic matter (FPOM) export remained below pre-treatment levels. The reduction in leaf standing crop in the treatment stream resulted in significant increases in FPOM (23), fine inorganic particulate (33), and gravel export (103). After small wood removal we observed significant increases in export of fine inorganic particulates (23) and gravel (73) from the treatment stream. A greater proportion of coarse and FBOM standing crop was exported from the treatment stream during the litter exclusion and small wood removal periods than from the reference stream. Following the addition of slow and mixed leaves this trend was reversed, demonstrating the importance of leaf standing crop in the retention of POM. Our long-term experiment demonstrates that the quantity and type of riparian inputs to forested headwater streams will affect POM standing crop and export of POM and sediments to downstream ecosystems, and that small wood is more critical to retaining sediments and POM in small streams than previously recognized.
National Science Foundation DEB-9207498, DEB-9629268, DEB-0212315
en_US
Ecological Society of America
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
allochthonous
Appalachia—streams
Coweeta hydrologic laboratory
North Carolina
detrital complexity
detritus
leaf litter
exclusion
organic matter standing crop
organic matter transport
sediment transport
seston
storms
Wood
food-web
mountain stream
forest-stream
Woody debris
terrestrial
dynamics
carbon
retention
litter
transport
Storage and export of organic matter in a headwater stream: responses to long-term detrital manipulations
Ecosphere
Article - Refereed
ORIGINAL
es12-00061%2E1.pdf
es12-00061%2E1.pdf
Published version
application/pdf
3393376
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es12-00061%2E1.pdf.txt
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77575
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10919/46814
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/46814
2023-11-17 20:34:04.377
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1112302022-07-14T07:12:34Zcom_10919_23829com_10919_5553col_10919_23830
Jackson-Blake, Leah A.
Clayer, Francois
de Eyto, Elvira
French, Andrew S.
Dolores Frias, Maria
Mercado-Bettin, Daniel
Moore, Tadhg
Puertolas, Laura
Poole, Russell
Rinke, Karsten
Shikhani, Muhammed
van der Linden, Leon
Marce, Rafael
2022-07-13T12:51:17Z
2022-07-13T12:51:17Z
2022-03-14
1027-5606
http://hdl.handle.net/10919/111230
https://doi.org/10.5194/hess-26-1389-2022
26
5
1607-7938
Advance warning of seasonal conditions has the potential to assist water management in planning and risk mitigation, with large potential social, economic, and ecological benefits. In this study, we explore the value of seasonal forecasting for decision-making at five case study sites located in extratropical regions. The forecasting tools used integrate seasonal climate model forecasts with freshwater impact models of catchment hydrology, lake conditions (temperature, water level, chemistry, and ecology), and fish migration timing and were co-developed together with water managers. To explore the decision-making value of forecasts, we carried out a qualitative assessment of (1) how useful forecasts would have been for a problematic past season and (2) the relevance of any windows of opportunity (seasons and variables where forecasts are thought to perform well) for management. Overall, water managers were optimistic about the potential for improved decision-making and identified actions that could be taken based on forecasts. However, there was often a mismatch between those variables that could best be predicted and those which would be most useful for management. Reductions in forecast uncertainty and a need to develop practical, hands-on experience were identified as key requirements before forecasts would be used in operational decision-making. Seasonal climate forecasts provided little added value to freshwater forecasts in these extratropical study sites, and we discuss the conditions under which seasonal climate forecasts with only limited skill are most likely to be worth incorporating into freshwater forecasting workflows.
MINECO-AEI [PCIN-2017062, PCIN-2017-092]; FORMAS; BMBF; EPA; RCN [274208]; IFD; European Union [690462]
Published version
This work was carried out as part of the WATExR project, part of ERA4CS, an ERA-NET initiated by JPI Climate and funded by MINECO-AEI (ES; grant nos. PCIN-2017062 and PCIN-2017-092), FORMAS (SE), BMBF (DE), EPA (IE), RCN (NO; grant no. 274208), and IFD (DK), with co-funding by the European Union (grant no. 690462).
application/pdf
en
Copernicus
Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
ensemble streamflow prediction
hydrologic prediction
system
skill
model
Opportunities for seasonal forecasting to support water management outside the tropics
Hydrology and Earth System Sciences
Article - Refereed
Text
ORIGINAL
hess-26-1389-2022.pdf
hess-26-1389-2022.pdf
Published version
application/pdf
1182838
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MD5
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hess-26-1389-2022.pdf.txt
hess-26-1389-2022.pdf.txt
Extracted text
text/plain
84206
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10919/111230
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/111230
2022-07-14 07:12:34.714
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/250112020-11-12T20:37:01Zcom_10919_23829com_10919_5553col_10919_23830
Cairns, John Jr.
Biological Sciences
2014-01-23T02:36:26Z
2014-01-23T02:36:26Z
2009
http://hdl.handle.net/10919/25011
http://www.johncairns.net/Papers/Two%20Estranged%20Cultures.pdf
In 1959, the British scientist C. P. Snow gave the prestigious Rede lecture with the thesis of a breakdown of communication between the "two cultures" of modern society- the sciences and the humanities-which was a major hindrance to solving the world's problems (Wikipedia). In the 21st century, a more serious breakdown has occurred between the two cultures of politicians and scientists, especially concerning the five critical, global problems:(1) climate, (2) acidification of the oceans, (3) overpopulaition, (4) ecological overshoot, (5) damage, possiblu irreversible, to the biospheric life support system. All these global problems require a holistic approach and will only be solved by exemplary collaboration among the world's nations, using a "top-down" global-system perspective. Special interest views are importatnt, but should not dominate. However, in the United States, and most other countries, special interest groups and lobbyists do dominate. The global system, Earth, has no well financed, politically powerful lobby. The result is that two, global, ecological tipping points have been passed and the damage is irresversible in timeframes of interest to humans. Can this alienation between scientists and politicians be resolved? It must be quickly resolved if civilization is to be saved.
en_US
Science and Society
In Copyright
http://rightsstatements.org/vocab/InC/1.0/
estrangement
alientation
politicians
scientists
two cultures
global collaboration
Two Estranged Cultures
Article
Text
ORIGINAL
cairns_two_estranged_cultures.pdf
application/pdf
33267
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MD5
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cairns_two_estranged_cultures.pdf.txt
cairns_two_estranged_cultures.pdf.txt
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text/plain
14266
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10919/25011
oai:vtw-prod.eed0.cloud.lib.vt.edu:10919/25011
2020-11-12 20:37:01.515
http://rightsstatements.org/vocab/InC/1.0/
In Copyright
open.access
https://vtw-prod.eed0.cloud.lib.vt.edu
VTechWorks
vtechworks@vt.edu
oai:vtechworks.lib.vt.edu:10919/1022242021-02-05T08:29:38Zcom_10919_23829com_10919_5553col_10919_23830
Goodchild, Christopher G.
Beck, Michelle L.
VanDiest, Issac
Czesak, Frankie N.
Lane, Samuel J.
Sewall, Kendra B.
Biological Sciences
2021-02-04T17:04:17Z
2021-02-04T17:04:17Z
2021-01-08
http://hdl.handle.net/10919/102224
https://doi.org/10.1016/j.ecoenv.2020.111850
210
Lead (Pb) is a pervasive global contaminant that interferes with sensitive windows for neurological development and causes oxidative damage to tissues. The effects of moderate and high exposure to Pb have been well-studied in birds, but whether low-level early-life exposure to Pb influences adult phenotype remains unclear. Female songbirds use a male’s song and coloration to discriminate between high- and low-quality males. Therefore, if early-life exposure to Pb disrupts song learning ability or shifts the allocation of antioxidant pigments away from colorful secondary sexual traits, male birds exposed to Pb may be less attractive to females. We exposed developing zebra finches (Taeniopygia guttata) to Pb-contaminated drinking water (100 or 1000 parts per billion [ppb]) after hatching (days 0–100). Once male finches reached adulthood (120–150 days post hatch), we measured song learning ability, coloration of bill and cheek patches, and volume of song nuclei in the brain. We also measured female preference for Pb-exposed males relative to control males. Finally, we measured motoric and spatial cognitive performance in male and female finches to assess whether cognitive traits differed in their sensitivity to Pb exposure. Male zebra finches exposed to 1000 ppb Pb had impaired song learning ability, reduced volume of song nuclei, bills with less redness and received less attention from females. Additionally, Pb exposure impaired motoric performance in both male and female finches but did not affect performance in a spatial cognitive task. Adult finches exposed to Pb-contaminated water had higher blood-Pb levels, though in all cases blood-Pb levels were below 7.0 μg dL⁻¹. This study suggests that low-level exposure to Pb contributes to cognitive deficits that persist into adulthood and may indirectly influence fitness by altering secondary sexual traits and reducing male attractiveness.
Virginia Tech Institute for Critical Technology and Applied Science
application/pdf
en
Elsevier
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
Dietary lead
Neurotoxicity
Song learning
Song nuclei
Bill coloration
Mate choice
Male zebra finches exposed to lead (Pb) during development have reduced volume of song nuclei, altered sexual traits, and received less attention from females as adults
Ecotoxicology and Environmental Safety
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