bluuebunny/crossref_metadata_2025_split huggingface dataset:
FTY720-induced human pulmonary endothelial barrier enhancement is mediated by c-Abl
Authors: Wang L. , Chiang E.T. , Simmons J.T. , Garcia J.G.N. , Dudek S.M. 
Published: November 2010

Strategies to improve pulmonary endothelial barrier function are needed to reverse the devastating effects of vascular leak in acute respiratory distress syndrome. FTY720 is a pharmaceutical analogue of the potent barrier-enhancing phospholipid sphingosine 1-phosphate (S1P). FTY720 decreases vascular permeability by an incompletely characterised mechanism that differs from S1P. Here, we describe its barrier-promoting effects on intracellular signalling and junctional assembly formation in human pulmonary endothelium.Permeability of cultured human pulmonary endothelial cells was assessed using transendothelial electrical resistance and dextran transwell assays. Junctional complex formation was assessed using membrane fractionation and immunofluorescence. Pharmacological inhibitors and small interfering (si)RNA were utilised to determine the effects of individual components on permeability.Unlike S1P, FTY720 failed to induce membrane translocation of adherens junction or tight junction proteins. β-catenin, occludin, claudin-5 or zona occludens protein (ZO)-1/ZO-2 siRNAs did not alter FTY720-induced barrier enhancement. FTY720 induced focal adhesion kinase (FAK) phosphorylation and focal adhesion formation, with FAK siRNA partially attenuating the prolonged phase of barrier enhancement. Inhibition of Src, protein kinase (PK)A, PKG, PKC or protein phosphatase 2A failed to alter FTY720-induced barrier enhancement. FTY720 increased c-Abl tyrosine kinase activity and c-Abl siRNA attenuated peak barrier enhancement after FTY720.FTY720 enhances endothelial barrier function by a novel pathway involving c-Abl signalling.
The oxidation induced by antimyeloperoxidase antibodies triggers fibrosis in microscopic polyangiitis
Authors: Guilpain P. , Chéreau C. , Goulvestre C. , Servettaz A. , Montani D. , Tamas N. , Pagnoux C. , Hachulla E. , Weill B. , Guillevin L. , Mouthon L. , Batteux F. 
Published: November 2010

Lung fibrosis is considered a severe manifestation of microscopic polyangiitis (MPA). Antimyeloperoxidase (anti-MPO) antibodies in MPA patients’ sera can activate MPO and lead to the production of reactive oxygen species (ROS). While high levels of ROS are cytotoxic, low levels can induce fibroblast proliferation. Therefore, we hypothesised that the oxidative stress induced by anti-MPO antibodies could contribute to lung fibrosis.24 MPA patients (45 sera) were enrolled in the study, including nine patients (22 sera) with lung fibrosis. Serum advanced oxidation protein products (AOPP), MPO-induced hypochlorous acid (HOCl) and serum-induced fibroblast proliferation were assayed.AOPP levels, MPO-induced HOCl production and serum-induced fibroblast proliferation were higher in patients than in healthy controls (p<0.0001, p = 0.0001 and p = 0.0005, respectively). Increased HOCl production was associated with active disease (p = 0.002). Serum AOPP levels and serum-induced fibroblast proliferation were higher in patients with active MPA and lung fibrosis (p<0.0001). A significant linear relationship between fibroblast proliferation, AOPP levels and HOCl production was observed only in patients with lung fibrosis.Oxidative stress, in particular the production of HOCl through the interaction of MPO with anti-MPO antibodies, could trigger the fibrotic process observed in MPA.
Epistatic QTL pairs associated with meat quality and carcass composition traits in a porcine Duroc × Pietrain population
Authors: Große-Brinkhaus Christine , Jonas Elisabeth , Buschbell Heiko , Phatsara Chirawath , Tesfaye Dawit , Jüngst Heinz , Looft Christian , Schellander Karl , Tholen Ernst 
Published: November 2010

AbstractBackgroundQuantitative trait loci (QTL) analyses in pig have revealed numerous individual QTL affecting growth, carcass composition, reproduction and meat quality, indicating a complex genetic architecture. In general, statistical QTL models consider only additive and dominance effects and identification of epistatic effects in livestock is not yet widespread. The aim of this study was to identify and characterize epistatic effects between common and novel QTL regions for carcass composition and meat quality traits in pig.MethodsFive hundred and eighty five F2pigs from a Duroc × Pietrain resource population were genotyped using 131 genetic markers (microsatellites and SNP) spread over the 18 pig autosomes. Phenotypic information for 26 carcass composition and meat quality traits was available for all F2animals. Linkage analysis was performed in a two-step procedure using a maximum likelihood approach implemented in the QxPak program.ResultsA number of interacting QTL was observed for different traits, leading to the identification of a variety of networks among chromosomal regions throughout the porcine genome. We distinguished 17 epistatic QTL pairs for carcass composition and 39 for meat quality traits. These interacting QTL pairs explained up to 8% of the phenotypic variance.ConclusionsOur findings demonstrate the significance of epistasis in pigs. We have revealed evidence for epistatic relationships between different chromosomal regions, confirmed known QTL loci and connected regions reported in other studies. Considering interactions between loci allowed us to identify several novel QTL and trait-specific relationships of loci within and across chromosomes.
Prenatal exposure of ethanol induces increased glutamatergic neuronal differentiation of neural progenitor cells
Authors: Kim Ki Chan , Go Hyo Sang , Bak Hae Rang , Choi Chang Soon , Choi Inha , Kim Pitna , Han Seol-Heui , Han So Min , Shin Chan Young , Ko Kwang Ho 
Published: November 2010

AbstractBackgroundPrenatal ethanol exposure during pregnancy induces a spectrum of mental and physical disorders called fetal alcohol spectrum disorder (FASD). The central nervous system is the main organ influenced by FASD, and neurological symptoms include mental retardation, learning abnormalities, hyperactivity and seizure susceptibility in childhood along with the microcephaly. In this study, we examined whether ethanol exposure adversely affects the proliferation of NPC and de-regulates the normal ratio between glutamatergic and GABAergic neuronal differentiation using primary neural progenitor culture (NPC) andin vivoFASD models.MethodsNeural progenitor cells were cultured from E14 embryo brain of Sprague-Dawley rat. Pregnant mice and rats were treated with ethanol (2 or 4 g/kg/day) diluted with normal saline from E7 to E16 forin vivoFASD animal models. Expression level of proteins was investigated by western blot analysis and immunocytochemical assays. MTT was used for cell viability. Proliferative activity of NPCs was identified by BrdU incorporation, immunocytochemistry and FACS analysis.ResultsReduced proliferation of NPCs by ethanol was demonstrated using BrdU incorporation, immunocytochemistry and FACS analysis. In addition, ethanol induced the imbalance between glutamatergic and GABAergic neuronal differentiationviatransient increase in the expression of Pax6, Ngn2 and NeuroD with concomitant decrease in the expression of Mash1. Similar pattern of expression of those transcription factors was observed using anin vivomodel of FASD as well as the increased expression of PSD-95 and decreased expression of GAD67.ConclusionsThese results suggest that ethanol induces hyper-differentiation of glutamatergic neuron through Pax6 pathway, which may underlie the hyper-excitability phenotype such as hyperactivity or seizure susceptibility in FASD patients.
EcoTILLING in Capsicum species: searching for new virus resistances
Authors: Ibiza Vicente P , Cañizares Joaquín , Nuez Fernando 
Published: November 2010

Abstract

Background
The EcoTILLING technique allows polymorphisms in target genes of natural populations to be quickly analysed or identified and facilitates the screening of genebank collections for desired traits. We have developed an EcoTILLING platform to exploit Capsicum genetic resources. A perfect example of the utility of this EcoTILLING platform is its application in searching for new virus-resistant alleles in Capsicum genus. Mutations in translation initiation factors (eIF4E, eIF(iso)4E, eIF4G and eIF(iso)4G) break the cycle of several RNA viruses without affecting the plant life cycle, which makes these genes potential targets to screen for resistant germplasm.

Results
We developed and assayed a cDNA-based EcoTILLING platform with 233 cultivated accessions of the genus Capsicum. High variability in the coding sequences of the eIF4E and eIF(iso)4E genes was detected using the cDNA platform. After sequencing, 36 nucleotide changes were detected in the CDS of eIF4E and 26 in eIF(iso)4E. A total of 21 eIF4E haplotypes and 15 eIF(iso)4E haplotypes were identified. To evaluate the functional relevance of this variability, 31 possible eIF4E/eIF(iso)4E combinations were tested against Potato virus Y. The results showed that five new eIF4E variants (pvr2

10
, pvr2

11
, pvr2

12
, pvr2

13
and pvr2

14
) were related to PVY-resistance responses.

Conclusions
EcoTILLING was optimised in different Capsicum species to detect allelic variants of target genes. This work is the first to use cDNA instead of genomic DNA in EcoTILLING. This approach avoids intronic sequence problems and reduces the number of reactions. A high level of polymorphism has been identified for initiation factors, showing the high genetic variability present in our collection and its potential use for other traits, such as genes related to biotic or abiotic stresses, quality or production. Moreover, the new eIF4E and eIF(iso)4E alleles are an excellent collection for searching for new resistance against other RNA viruses.
Single-cell qPCR on dispersed primary pituitary cells -an optimized protocol
Authors: Hodne Kjetil , Haug Trude M , Weltzien Finn-Arne 
Published: November 2010

Abstract

Background
The incidence of false positives is a potential problem in single-cell PCR experiments. This paper describes an optimized protocol for single-cell qPCR measurements in primary pituitary cell cultures following patch-clamp recordings. Two different cell harvesting methods were assessed using both the GH4 prolactin producing cell line from rat, and primary cell culture from fish pituitaries.

Results
Harvesting whole cells followed by cell lysis and qPCR performed satisfactory on the GH4 cell line. However, harvesting of whole cells from primary pituitary cultures regularly produced false positives, probably due to RNA leakage from cells ruptured during the dispersion of the pituitary cells. To reduce RNA contamination affecting the results, we optimized the conditions by harvesting only the cytosol through a patch pipette, subsequent to electrophysiological experiments. Two important factors proved crucial for reliable harvesting. First, silanizing the patch pipette glass prevented foreign extracellular RNA from attaching to charged residues on the glass surface. Second, substituting the commonly used perforating antibiotic amphotericin B with β-escin allowed efficient cytosol harvest without loosing the giga seal. Importantly, the two harvesting protocols revealed no difference in RNA isolation efficiency.

Conclusion
Depending on the cell type and preparation, validation of the harvesting technique is extremely important as contaminations may give false positives. Here we present an optimized protocol allowing secure harvesting of RNA from single cells in primary pituitary cell culture following perforated whole cell patch clamp experiments.
The use of opioids at the end of life: the knowledge level of Dutch physicians as a potential barrier to effective pain management
Authors: Rurup Mette L , Rhodius Christiaan A , Borgsteede Sander D , Boddaert Manon SA , Keijser Astrid GM , Pasman H Roeline W , Onwuteaka-Philipsen Bregje D 
Published: November 2010

Abstract

Background
Pain is still one of the most frequently occurring symptoms at the end of life, although it can be treated satisfactorily in most cases if the physician has adequate knowledge. In the Netherlands, almost 60% of the patients with non-acute illnesses die at home where end of life care is coordinated by the general practitioner (GP); about 30% die in hospitals (cared for by clinical specialists), and about 10% in nursing homes (cared for by elderly care physicians).
The research question of this study is: what is the level of knowledge of Dutch physicians concerning pain management and the use of opioids at the end of life?

Methods
A written questionnaire was sent to a random sample of physicians of specialties most often involved in end of life care in the Netherlands. The questionnaire was completed by 406 physicians, response rate 41%.

Results
Almost all physicians were aware of the most basal knowledge about opioids, e.g. that it is important for treatment purposes to distinguish nociceptive from neuropathic pain (97%). Approximately half of the physicians (46%) did not know that decreased renal function raises plasma concentration of morphine(-metabolites) and 34% of the clinical specialists erroneously thought opioids are the favoured drug for palliative sedation.
Although 91% knew that opioids titrated against pain do not shorten life, 10% sometimes or often gave higher dosages than needed with the explicit aim to hasten death. About half felt sometimes or often pressured by relatives to hasten death by increasing opioiddosage.
The large majority (83%) of physicians was interested in additional education about subjects related to the end of life, the most popular subject was opioid rotation (46%).

Conclusions
Although the basic knowledge of physicians was adequate, there seemed to be a lack of knowledge in several areas, which can be a barrier for good pain management at the end of life. From this study four areas emerge, in which it seems likely that an improvement can improve the quality of pain management at the end of life for many patients in the Netherlands: 1)palliative sedation; 2)expected effect of opioids on survival; and 3) opioid rotation.
Effects of heliox as carrier gas on ventilation and oxygenation in an animal model of piston-type HFOV: a crossover experimental study
Authors: Zeynalov Bakhtiyar , Hiroma Takehiko , Nakamura Tomohiko 
Published: November 2010

Abstract

Objective
This study aimed to compare gas exchange with heliox and oxygen-enriched air during piston-type high-frequency oscillatory ventilation (HFOV). We hypothesized that helium gas would improve both carbon dioxide elimination and arterial oxygenation during piston-type HFOV.

Method
Five rabbits were prepared and ventilated by piston-type HFOV with carrier 50% helium/oxygen (heliox50) or 50% oxygen/nitrogen (nitrogen50) gas mixture in a crossover study. Changing the gas mixture from nitrogen50 to heliox50 and back was performed five times per animal with constant ventilation parameters. Arterial blood gas, vital function and respiratory test indices were recorded.

Results
Compared with nitrogen50, heliox50 did not change PaCO2 when stroke volume remained constant, but significantly reduced PaCO2 after alignment of amplitude pressure. No significant changes in PaO2 were seen despite significant decreases in mean airway pressure with heliox50 compared with nitrogen50.

Conclusion
This study demonstrated that heliox enhances CO2 elimination and maintains oxygenation at the same amplitude but with lower airway pressure compared to air/O2 mix gas during piston-type HFOV.
The effect of body size on the wing movements of pteropodid bats, with insights into thrust and lift production
Authors: Riskin Daniel K. , Iriarte-Díaz José , Middleton Kevin M. , Breuer Kenneth S. , Swartz Sharon M. 
Published: November 2010

SUMMARYIn this study we compared the wing kinematics of 27 bats representing six pteropodid species ranging more than 40 times in body mass (Mb=0.0278–1.152 kg), to determine whether wing posture and overall wing kinematics scaled as predicted according to theory. The smallest species flew in a wind tunnel and the other five species in a flight corridor. Seventeen kinematic markers on the midline and left side of the body were tracked in three dimensions. We used phylogenetically informed reduced major axis regression to test for allometry. We found that maximum wingspan (bmax) and maximum wing area (Smax) scaled with more positive allometry, and wing loading (Qs) with more negative allometry (bmax∝Mb0.423; Smax∝Mb0.768; Qs∝Mb0.233) than has been reported in previous studies that were based on measurements from specimens stretched out flat on a horizontal surface. Our results suggest that larger bats open their wings more fully than small bats do in flight, and that for bats, body measurements alone cannot be used to predict the conformation of the wings in flight. Several kinematic variables, including downstroke ratio, wing stroke amplitude, stroke plane angle, wing camber and Strouhal number, did not change significantly with body size, demonstrating that many aspects of wing kinematics are similar across this range of body sizes. Whereas aerodynamic theory suggests that preferred flight speed should increase with mass, we did not observe an increase in preferred flight speed with mass. Instead, larger bats had higher lift coefficients (CL) than did small bats (CL∝Mb0.170). Also, the slope of the wingbeat period (T) to body mass regression was significantly more shallow than expected under isometry (T∝Mb0.180), and angle of attack (α) increased significantly with body mass [α∝log(Mb)7.738]. None of the bats in our study flew at constant speed, so we used multiple regression to isolate the changes in wing kinematics that correlated with changes in flight speed, horizontal acceleration and vertical acceleration. We uncovered several significant trends that were consistent among species. Our results demonstrate that for medium- to large-sized bats, the ways that bats modulate their wing kinematics to produce thrust and lift over the course of a wingbeat cycle are independent of body size.
The effect of fin ray flexural rigidity on the propulsive forces generated by a biorobotic fish pectoral fin
Authors: Tangorra James L. , Lauder George V. , Hunter Ian W. , Mittal Rajat , Madden Peter G. A. , Bozkurttas Meliha 
Published: November 2010

SUMMARY
A biorobotic pectoral fin was developed and used to study how the flexural rigidities of fin rays within a highly deformable fish fin affect the fin's propulsive forces. The design of the biorobotic fin was based on a detailed analysis of the pectoral fin of the bluegill sunfish (Lepomis macrochirus). The biorobotic fin was made to execute the kinematics used by the biological fin during steady swimming, and to have structural properties that modeled those of the biological fin. This resulted in an engineered fin that had a similar interaction with the water as the biological fin and that created close approximations of the three-dimensional motions, flows, and forces produced by the sunfish during low speed, steady swimming. Experimental trials were conducted during which biorobotic fins of seven different stiffness configurations were flapped at frequencies from 0.5 to 2.0 Hz in flows with velocities that ranged from 0 to 270 mm s–1. During these trials, thrust and lift forces were measured, kinematics were recorded in three dimensions, and digital particle image velocimetry was used to evaluate flow hydrodynamics. The results of the trials revealed that slight changes to the fin's mechanical properties or to the operating conditions can have significant impact on the direction, magnitude and time course of the propulsive forces. In general, the magnitude of the 2-D (thrust and lift) propulsive force scaled with fin ray stiffness, and increased as the fin's flapping speed increased or as the velocity of the flow decreased.
Sequence Capture and Next-Generation Resequencing of Multiple Tagged Nucleic Acid Samples for Mutation Screening of Urea Cycle Disorders
Authors: Amstutz Ursula , Andrey-Zürcher Gisela , Suciu Dominic , Jaggi Rolf , Häberle Johannes , Largiadèr Carlo R 
Published: November 2010

BACKGROUND
Molecular genetic testing is commonly used to confirm clinical diagnoses of inherited urea cycle disorders (UCDs); however, conventional mutation screenings encompassing only the coding regions of genes may not detect disease-causing mutations occurring in regulatory elements and introns. Microarray-based target enrichment and next-generation sequencing now allow more-comprehensive genetic screening. We applied this approach to UCDs and combined it with the use of DNA bar codes for more cost-effective, parallel analyses of multiple samples.

METHODS
We used sectored 2240-feature medium-density oligonucleotide arrays to capture and enrich a 199-kb genomic target encompassing the complete genomic regions of 3 urea cycle genes, OTC (ornithine carbamoyltransferase), CPS1 (carbamoyl-phosphate synthetase 1, mitochondrial), and NAGS (N-acetylglutamate synthase). We used the Genome Sequencer FLX System (454 Life Sciences) to jointly analyze 4 samples individually tagged with a 6-bp DNA bar code and compared the results with those for an individually sequenced sample.

RESULTS
Using a low tiling density of only 1 probe per 91 bp, we obtained strong enrichment of the targeted loci to achieve ≥90% coverage with up to 64% of the sequences covered at a sequencing depth ≥10-fold. We observed a very homogeneous sequence representation of the bar-coded samples, which yielded a &gt;30% increase in the sequence data generated per sample, compared with an individually processed sample. Heterozygous and homozygous disease-associated mutations were correctly detected in all samples.

CONCLUSIONS
The use of DNA bar codes and the use of sectored oligonucleotide arrays for target enrichment enable parallel, large-scale analysis of complete genomic regions for multiple genes of a disease pathway and for multiple samples simultaneously. This approach thus may provide an efficient tool for comprehensive diagnostic screening of mutations.
Is it possible to avoid hypopituitarism after irradiation of pituitary adenomas by the Leksell gamma knife?
Authors: Marek Josef , Ježková Jana , Hána Václav , Kršek Michal , Bandúrová L'ubomíra , Pecen Ladislav , Vladyka Vilibald , Liščák Roman 
Published: November 2010

ObjectiveRadiation therapy is one of the treatment options for pituitary adenomas. The most common side effect associated with Leksell gamma knife (LGK) irradiation is the development of hypopituitarism. The aim of this study was to verify that hypopituitarism does not develop if the maximum mean dose to pituitary is kept under 15 Gy and to evaluate the influence of maximum distal infundibulum dose on the development of hypopituitarism.Design and methodsWe followed the incidence of hypopituitarism in 85 patients irradiated with LGK in 1993–2003. The patients were divided in two subgroups: the first subgroup followed prospectively (45 patients), irradiated with a mean dose to pituitary <15 Gy; the second subgroup followed retrospectively 1993–2001 and prospectively 2001–2009 (40 patients), irradiated with a mean dose to pituitary >15 Gy. Serum TSH, free thyroxine, testosterone or 17β-oestradiol, IGF1, prolactin and cortisol levels were evaluated before and every 6 months after LGK irradiation.ResultsHypopituitarism after LGK irradiation developed only in 1 out of 45 (2.2%) patients irradiated with a mean dose to pituitary <15 Gy, in contrast to 72.5% patients irradiated with a mean dose to pituitary >15 Gy. The radiation dose to the distal infundibulum was found as an independent factor of hypopituitarism with calculated maximum safe dose of 17 Gy.ConclusionKeeping the mean radiation dose to pituitary under 15 Gy and the dose to the distal infundibulum under 17 Gy prevents the development of hypopituitarism following LGK irradiation.