106 research outputs found
CHROMIC AND IRON OXIDES AS FECAL MARKERS TO IDENTIFY INDIVIDUAL WHOOPING CRANES
The whooping crane (Grus americana) is listed as endangered under the IUCN Red List, the United States Endangered Species Act, and the Canadian Species at Risk Act (BirdLife International 2012, CWS and USFWS 2007). A major focus of recovery efforts for this endangered species is reintroduction to establish new populations (CWS and USFWS 2007). Captive populations are critical as a source of individuals for reintroduction efforts and also serve as insurance populations. Currently, there are a total of 157 whooping cranes held in captive breeding centers across North America, with the largest at the USGS Patuxent Wildlife Research Center (PWRC) in Laurel, Maryland. Birds produced in this facility are currently being released as part of efforts to establish the Eastern Migratory Population (EMP, Urbanek et al. 2005) and in an effort to establish a non-migratory population in Louisiana. In the past decade, PWRC has produced and released annually an average of 18 birds into the wild; however, reproductive performance of birds at this facility is lower than desired. PWRC had a 60% fertility rate for eggs laid from 2000 through 2010 (J. N. Chandler, personal communication, 2011). Furthermore, reproductive onset in this captive population appears to be delayed compared to wild populations. In wild populations, reproductive onset (production of sperm and eggs) normally occurs ~5 years of age in both males and females, ~2 years after initial pair formation occurs (Ellis et al., 1996), while some females in the EMP have laid eggs earlier than 5 years of age (Converse et al. 2011). However, PWRC females in some cases do not start to lay eggs until 7 years of age (Mirande et al. 1996). Currently, the PWRC population consists of a total of 74 whooping cranes, including 22 pairs. Six of these pairs (27%) are consistently infertile (i.e., no production of fertile eggs) and 3 other pairs (14%) have low fertility (30- 45% fertility in eggs laid), which is variable from year to year. Six pairs (27%) are recently formed and have not produced eggs, and so have unknown fertility. This leaves only 7 pairs (33%) which contribute maximally to PWRCâs chick production (J. N. Chandler, personal communication, 2011). Because of the challenges occurring within this captive colony, PWRC and Smithsonian National Zoo have initiated a joint research project to identify potential underlying causes of poor reproduction in captive whooping cranes
Genome-Wide RNA Sequencing of Human Trabecular Meshwork Cells Treated with TGF-ÎČ1: Relevance to Pseudoexfoliation Glaucoma
Pseudoexfoliation glaucoma (XFG) is an aggressive form of secondary open angle glaucoma, characterised by the production of exfoliation material and is estimated to affect 30 million people worldwide. Activation of the TGF-β pathway by TGF-β1 has been implicated in the pathogenesis of pseudoexfoliation glaucoma. To further investigate the role of TGF-β1 in glaucomatous changes in the trabecular meshwork (TM), we used RNA-Seq to determine TGF-β1 induced changes in the transcriptome of normal human trabecular meshwork (HTM) cells. The main purpose of this study was to perform a hypothesis-independent RNA sequencing analysis to investigate genome-wide alterations in the transcriptome of normal HTMs stimulated with TGF-β1 and investigate possible pathophysiological mechanisms driving XFG. Our results identified multiple differentially expressed genes including several genes known to be present in exfoliation material. Significantly altered pathways, biological processes and molecular functions included extracellular matrix remodelling, Hippo and Wnt pathways, the unfolded protein response, oxidative stress, and the antioxidant system. This cellular model of pseudoexfoliation glaucoma can provide insight into disease pathogenesis and support the development of novel therapeutic interventions
Impact of hypercapnia on alveolar Na+-transport : Establishing a system for ENaC-protein detection
Acute respiratory distress syndrome is a life threatening condition triggered by a variety of
pulmonary and extrapulmonary causes, that is characterized by pulmonary edema and
subsequently impaired gas exchange. Due to lung protective ventilation strategies, its
treatment is often associated with systemic accumulation of CO2, a condition termed
permissive hypercapnia. Recent studies report a negative effect of CO2 on alveolar fluid
clearance, a process mediated by its two key elements the Na+,K+-ATPase and epithelial
Na+-channels (ENaCs). A reduced activity of the Na+,K+-ATPase during hypercapnia has
already been demonstrated, but regulation of ENaC has never been directly linked to CO2.
Many molecular signaling events that are activated during hypercapnia are known to
regulate ENaC function, so the present study aimed to generate and subsequently apply
techniques to investigate a possible contribution of ENaC to the reduction of alveolar
epithelial fluid transport upon hypercapnia.
ENaC function was studied in H441 cells by Ussing chamber experiments which revealed
no significant regulation during short term hypercapnia, but a clear reduction of ENaC
function during sustained hypercapnia.
To identify the signaling mechanism on the molecular level, epitope-tagged human ENaC
constructs for the α-, β- and γ-subunit were cloned and initially expressed in A549 cells.
Exposition to hypercapnia up to 4 hours did not significantly reduce cell surface expression
of the ENaC-subunits, but after 24 hours, a significant decrease of β-ENaC was observed.
Since the molecular sizes of α- and γ-ENaC expressed in A549 cells were differing from
previously published studies, transfection of ENaC was continued in other cells. H441 cells
are commonly used for ENaC studies, so their transfection was established, yielding an
efficiency of about 60 %. The molecular sizes of transfected ENaC subunits matched the
pattern that was expected, but expression levels were evanescent and too low for further
experiments. Since ENaC detection in these two cell lines remained problematic, a novel
methodology was applied. Since the primary site of ENaC expression in the lung are
epithelial cells, rat primary alveolar epithelial cells type II were used as recipients for
ENaC plasmids. Non-viral transfection of ATII cells has been inefficient in the past, but
during the present study a protocol was generated to efficiently deliver nucleic acids to
exactly this cell type. ENaC expression was largely increased in ATII cells, compared to
the cell lines used, indicating that established system might be extremely useful for further
studies involving ENaC turnover.
Thus, a new and highly relevant, non-viral transfection technique for primary alveolar
epithelial type II cells was established, providing ground-breaking opportunities for future
pulmonary research.Das Atemnotsyndrom des Erwachsenen ist eine lebensbedrohliche Erkrankung, ausgelöst
durch eine Reihe von Faktoren, die direkt oder indirekt auf die Lunge einwirken .
Charakteristisch fĂŒr dieses Syndrom sind pulmonare Ădeme und daraus resultierend ein
eingeschrĂ€nkter Gasaustausch. Die daher benötigte kĂŒnstliche Beatmung fĂŒhrt im Zuge
von protektiven Beatmungsstrategien oft zu einer systemischen Anreicherung von CO2
(Hyperkapnie). Einige Studien zeigen, dass erhöhte CO2-Level den FlĂŒssigkeitstransport
der Lunge einschrĂ€nken. Dieser aktive Prozess wird maĂgeblich durch zwei Komponenten,
die Na+,K+-ATPase und epitheliale Na+-KanÀle (ENaCs), kontrolliert. Eine
BeeintrĂ€chtigung der Na+,K+-ATPase durch CO2 gezeigt, fĂŒr ENaCs ist dies bislang nicht
bekannt. Einige bekannte Regulatoren von ENaCs werden jedoch wÀhrend Hyperkapnie
aktiviert. Das Ziel der vorliegenden Arbeit war, Methoden zu etablieren und anzuwenden,
die einen möglichen Einfluss von CO2 auf ENaC zeigen.
Funktionelle Versuche wurden an H441-Zellen mit Ussing-Kammer-Messungen
durchgefĂŒhrt. WĂ€hrend akuter Hyperkapnie konnte keine signifikante Regulation von
ENaC nachgewiesen werden, jedoch war die ENaC-Funktion bei anhaltender Hyperkapnie
deutlich verringert.
Um die Signalwege auf molekularer Ebene zu untersuchen, wurde die α-, β- und γ-
Untereinheit des humanen ENaC kloniert, genetisch modifiziert und in A549 Zellen
ĂŒberexprimiert. Nach bis zu vierstĂŒndiger Hyperkapnie erfolgte keine Regulation von
ENaC, jedoch wurde nach 24 Stunden eine deutlich verminderte Menge β-ENaC in der
Zellmembran nachgewiesen. Da die GröĂen von α- und γ-ENaC von den bisher
publizierten abwichen, wurden weitere Versuche in H441 Zellen durchgefĂŒhrt. Die
Transfektion dieser Zelllinie wurde etabliert und erreichte eine Effizienz von ungefÀhr 60
%. Die posttranslationale Regulation der α- und γ-Untereinheiten, insbesondere die
proteolytische Aktivierung funktionierten wie in der Literatur beschrieben, jedoch waren
die Expressionslevel zu gering fĂŒr weitere Versuche. In der Lunge werden ENaCs
ĂŒberwiegend in epithelialen Zellen exprimiert. Diese Zellen konnten bisher jedoch nicht
effizient transfiziert werden, ohne Viren einzusetzen. In der vorliegenden Arbeit wurde
jedoch eine effiziente Methode zur Transfektion von primÀren epithelialen Zellen der Ratte
erarbeitet. Die Expression von transfizierten ENaC-Untereinheiten war in diesen Zellen
deutlich erhöht, weswegen die Etablierung dieses Systems ausschlaggebend fĂŒr weitere
Versuche ist.
Die vorliegende Arbeit beschreibt daher zum ersten Mal die nicht-virale, effiziente
Transfektion von primÀren alveolaren Zellen und liefert damit ein bedeutendes neues
Werkzeug fĂŒr die Lungenforschung
Planetary Candidates Observed by Kepler. VIII. A Fully Automated Catalog With Measured Completeness and Reliability Based on Data Release 25
We present the Kepler Object of Interest (KOI) catalog of transiting
exoplanets based on searching four years of Kepler time series photometry (Data
Release 25, Q1-Q17). The catalog contains 8054 KOIs of which 4034 are planet
candidates with periods between 0.25 and 632 days. Of these candidates, 219 are
new and include two in multi-planet systems (KOI-82.06 and KOI-2926.05), and
ten high-reliability, terrestrial-size, habitable zone candidates. This catalog
was created using a tool called the Robovetter which automatically vets the
DR25 Threshold Crossing Events (TCEs, Twicken et al. 2016). The Robovetter also
vetted simulated data sets and measured how well it was able to separate TCEs
caused by noise from those caused by low signal-to-noise transits. We discusses
the Robovetter and the metrics it uses to sort TCEs. For orbital periods less
than 100 days the Robovetter completeness (the fraction of simulated transits
that are determined to be planet candidates) across all observed stars is
greater than 85%. For the same period range, the catalog reliability (the
fraction of candidates that are not due to instrumental or stellar noise) is
greater than 98%. However, for low signal-to-noise candidates between 200 and
500 days around FGK dwarf stars, the Robovetter is 76.7% complete and the
catalog is 50.5% reliable. The KOI catalog, the transit fits and all of the
simulated data used to characterize this catalog are available at the NASA
Exoplanet Archive.Comment: 61 pages, 23 Figures, 9 Tables, Accepted to The Astrophysical Journal
Supplement Serie
Working memory training and brain structure and function in extremely preterm or extremely low birth weight children.
This study in children born extremely preterm (EP; <28âweeks' gestational age) or extremely low birth weight (ELBW; <1,000âg) investigated whether adaptive working memory training using CogmedÂź is associated with structural and/or functional brain changes compared with a placebo program. Ninety-one EP/ELBW children were recruited at a mean (standard deviation) age of 7.8 (0.4) years. Children were randomly allocated to Cogmed or placebo (45-min sessions, 5âdays a week over 5-7âweeks). A subset had usable magnetic resonance imaging (MRI) data pretraining and 2âweeks posttraining (structural, n = 48; diffusion, n = 43; task-based functional, n = 18). Statistical analyses examined whether cortical morphometry, white matter microstructure and blood oxygenation level-dependent (BOLD) signal during an n-back working memory task changed from pretraining to posttraining in the Cogmed and placebo groups separately. Interaction analyses between time point and group were then performed. There was a significant increase in neurite density in several white matter regions from pretraining to posttraining in both the Cogmed and placebo groups. BOLD signal in the posterior cingulate and precuneus cortices during the n-back task increased from pretraining to posttraining in the Cogmed but not placebo group. Evidence for group-by-time interactions for the MRI measures was weak, suggesting that brain changes generally did not differ between Cogmed and placebo groups. Overall, while some structural and functional MRI changes between the pretraining and posttraining period in EP/ELBW children were observed, there was little evidence of training-induced neuroplasticity, with changes generally identified in both groups. Trial registration Australian New Zealand Clinical Trials Registry, anzctr.org.au; ACTRN12612000124831
Acoustic sequences in non-human animals: a tutorial review and prospectus.
Animal acoustic communication often takes the form of complex sequences, made up of multiple distinct acoustic units. Apart from the well-known example of birdsong, other animals such as insects, amphibians, and mammals (including bats, rodents, primates, and cetaceans) also generate complex acoustic sequences. Occasionally, such as with birdsong, the adaptive role of these sequences seems clear (e.g. mate attraction and territorial defence). More often however, researchers have only begun to characterise - let alone understand - the significance and meaning of acoustic sequences. Hypotheses abound, but there is little agreement as to how sequences should be defined and analysed. Our review aims to outline suitable methods for testing these hypotheses, and to describe the major limitations to our current and near-future knowledge on questions of acoustic sequences. This review and prospectus is the result of a collaborative effort between 43 scientists from the fields of animal behaviour, ecology and evolution, signal processing, machine learning, quantitative linguistics, and information theory, who gathered for a 2013 workshop entitled, 'Analysing vocal sequences in animals'. Our goal is to present not just a review of the state of the art, but to propose a methodological framework that summarises what we suggest are the best practices for research in this field, across taxa and across disciplines. We also provide a tutorial-style introduction to some of the most promising algorithmic approaches for analysing sequences. We divide our review into three sections: identifying the distinct units of an acoustic sequence, describing the different ways that information can be contained within a sequence, and analysing the structure of that sequence. Each of these sections is further subdivided to address the key questions and approaches in that area. We propose a uniform, systematic, and comprehensive approach to studying sequences, with the goal of clarifying research terms used in different fields, and facilitating collaboration and comparative studies. Allowing greater interdisciplinary collaboration will facilitate the investigation of many important questions in the evolution of communication and sociality.This review was developed at an investigative workshop, âAnalyzing Animal Vocal Communication Sequencesâ that took place on October 21â23 2013 in Knoxville, Tennessee, sponsored by the National Institute for Mathematical and Biological Synthesis (NIMBioS). NIMBioS is an Institute sponsored by the National Science Foundation, the U.S. Department of Homeland Security, and the U.S. Department of Agriculture through NSF Awards #EF-0832858 and #DBI-1300426, with additional support from The University of Tennessee, Knoxville. In addition to the authors, Vincent Janik participated in the workshop. D.T.B.âs research is currently supported by NSF DEB-1119660. M.A.B.âs research is currently supported by NSF IOS-0842759 and NIH R01DC009582. M.A.R.âs research is supported by ONR N0001411IP20086 and NOPP (ONR/BOEM) N00014-11-1-0697. S.L.DeR.âs research is supported by the U.S. Office of Naval Research. R.F.-i-C.âs research was supported by the grant BASMATI (TIN2011-27479-C04-03) from the Spanish Ministry of Science and Innovation. E.C.G.âs research is currently supported by a National Research Council postdoctoral fellowship. E.E.V.âs research is supported by CONACYT, Mexico, award number I010/214/2012.This is the accepted manuscript. The final version is available at http://dx.doi.org/10.1111/brv.1216
Genome sequences of four cluster P mycobacteriophages
Four bacteriophages infecting Mycobacterium smegmatis mc2155 (three belonging to subcluster P1 and one belonging to subcluster P2) were isolated from soil and sequenced. All four phages are similar in the left arm of their genomes, but the P2 phage differs in the right arm. All four genomes contain features of temperate phages
Bone Marrow-Derived Progenitor Cells Augment Venous Remodeling in a Mouse Dorsal Skinfold Chamber Model
The delivery of bone marrow-derived cells (BMDCs) has been widely used to stimulate angiogenesis and arteriogenesis. We identified a progenitor-enriched subpopulation of BMDCs that is able to augment venular remodeling, a generally unexplored area in microvascular research. Two populations of BMDCs, whole bone marrow (WBM) and Linâ/Sca-1+ progenitor cells, were encapsulated in sodium alginate and delivered to a mouse dorsal skinfold chamber model. Upon observation that encapsulated Sca-1+ progenitor cells enhance venular remodeling, the cells and tissue were analyzed on structural and molecular levels. Venule walls were thickened and contained more nuclei after Sca-1+ progenitor cell delivery. In addition, progenitors expressed mRNA transcript levels of chemokine (C-X-C motif) ligand 2 (CXCL2) and interferon gamma (IFNÎł) that are over 5-fold higher compared to WBM. Tissues that received progenitors expressed significantly higher protein levels of vascular endothelial growth factor (VEGF), monocyte chemotactic protein-1 (MCP-1), and platelet derived growth factor-BB (PDGF-BB) compared to tissues that received an alginate control construct. Nine days following cell delivery, tissue from progenitor recipients contained 39% more CD45+ leukocytes, suggesting that these cells may enhance venular remodeling through the modulation of the local immune environment. Results show that different BMDC populations elicit different microvascular responses. In this model, Sca-1+ progenitor cell-derived CXCL2 and IFNÎł may mediate venule enlargement via modulation of the local inflammatory environment
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