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Article: Neonatal Impedance Cardiography in Asphyxiated Piglets-A Feasibility Study.

Berisha, Gazmend / Solberg, Rønnaug / Klingenberg, Claus / Solevåg, Anne Lee

2022 Volume 10, Page(s) 804353

Abstract: Objectives: Impedance cardiography (ICG) is a non-invasive method for continuous cardiac output measurement and has the potential to improve monitoring and treatment of sick neonates. PhysioFlow: Methods: Fifteen piglets, under continuous arterial ... ...

Abstract	Objectives: Impedance cardiography (ICG) is a non-invasive method for continuous cardiac output measurement and has the potential to improve monitoring and treatment of sick neonates. PhysioFlow Methods: Fifteen piglets, under continuous arterial heart rate (HR) and blood pressure (BP) monitoring, were asphyxiated until asystole. Cardiopulmonary resuscitation was performed and the piglets monitored after return of spontaneous circulation (ROSC). Arterial lactate was measured at baseline, every 5 min throughout asphyxiation, at asystole, and at 10 min and later every 30 min after ROSC. PhysioFlow Results: The piglets were asphyxiated for median (interquartile range) 30 (20-35) min and had a lactate at asystole of 15.0 (9.1-17.0) mmol/L. Out of a total of 20.991 registrations in all animals combined, there were 10.148 (48.3%) registrations with a signal quality ≥ 75%. Signal quality ≥ 75% varied in individual piglets from 7 to 82% of registrations. We analyzed 1.254 registrations recorded 30 min from initiation of asphyxia, i.e., in piglets with brief asphyxia times, this included cardiopulmonary resuscitation and post-ROSC observation. There was a positive correlation between CI and SVI ( Conclusion: About half of ICG-registrations in asphyxiated piglets were of good quality. However, signal quality was highly variable between piglets. In total, there was a higher proportion of reliable ICG-registrations than reported from clinical delivery room studies using electrical velocimetry. Our data are physiologically plausible and supports further research evaluating PhysioFlow
Language	English
Publishing date	2022-02-25
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2711999-3
ISSN	2296-2360
ISSN	2296-2360
DOI	10.3389/fped.2022.804353
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Article ; Online: Correct treatment for tongue-tie in infants.

Holmsen, Solveig Thorp / Lona, Ann-Magrit / Solberg, Rønnaug

Tidsskrift for den Norske laegeforening : tidsskrift for praktisk medicin, ny raekke

2021 Volume 141

Title translation	Riktig behandling for stramt tungebånd hos spedbarn.
MeSH term(s)	Ankyloglossia ; Breast Feeding ; Female ; Humans ; Infant ; Tongue
Language	Norwegian
Publishing date	2021-08-26
Publishing country	Norway
Document type	Journal Article
ZDB-ID	603504-8
ISSN	0807-7096 ; 0029-2001
ISSN (online)	0807-7096
ISSN	0029-2001
DOI	10.4045/tidsskr.21.0520
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Article: Long Non-Coding RNAs in Hypoxia and Oxidative Stress: Novel Insights Investigating a Piglet Model of Perinatal Asphyxia.

Grebstad Tune, Benedicte / Melheim, Maria / Åsegg-Atneosen, Monica / Dotinga, Baukje / Saugstad, Ola Didrik / Solberg, Rønnaug / Baumbusch, Lars Oliver

Biology

2023 Volume 12, Issue 4

Abstract: Birth asphyxia is the leading cause of death and disability in young children worldwide. Long non-coding RNAs (lncRNAs) may provide novel targets and intervention strategies due to their regulatory potential, as demonstrated in various diseases and ... ...

Abstract	Birth asphyxia is the leading cause of death and disability in young children worldwide. Long non-coding RNAs (lncRNAs) may provide novel targets and intervention strategies due to their regulatory potential, as demonstrated in various diseases and conditions. We investigated cardinal lncRNAs involved in oxidative stress, hypoxia, apoptosis, and DNA damage using a piglet model of perinatal asphyxia. A total of 42 newborn piglets were randomized into 4 study arms: (1) hypoxia-normoxic reoxygenation, (2) hypoxia-3 min of hyperoxic reoxygenation, (3) hypoxia-30 min of hyperoxic reoxygenation, and (4) sham-operated controls. The expression of lncRNAs
Language	English
Publishing date	2023-04-04
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2661517-4
ISSN	2079-7737
ISSN	2079-7737
DOI	10.3390/biology12040549
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Article ; Online: Gene expression in the intestine of newborn piglets after hypoxia-reoxygenation.

Dotinga, Baukje M / Bao, Mian / Solberg, Rønnaug / Saugstad, Ola D / Hulscher, Jan B F / Bos, Arend F / Plösch, Torsten / Kooi, Elisabeth M W

Pediatric research

2023 Volume 94, Issue 4, Page(s) 1365–1372

Abstract: Background: In preterm infants, intestinal hypoxia may partly contribute to the pathophysiology of necrotizing enterocolitis through changes in gene expression. Splanchnic hypoxia can be detected with monitoring of regional splanchnic oxygen saturation ( ...

Abstract	Background: In preterm infants, intestinal hypoxia may partly contribute to the pathophysiology of necrotizing enterocolitis through changes in gene expression. Splanchnic hypoxia can be detected with monitoring of regional splanchnic oxygen saturation (r Methods: Forty-two newborn piglets were randomized to control or intervention groups. Intervention groups were subjected to hypoxia until they were acidotic and hypotensive. Next, they were reoxygenated for 30 min according to randomization, i.e., 21% O Results: The expression of selected genes was not significantly different between control and intervention groups. No associations between mean r Conclusion: Our study suggests that hypoxia and reoxygenation cause reduced vascular adaptability, which seems to be associated with the upregulation of apoptosis and downregulation of inflammation. Impact: Our results provide important insight into the (patho)physiological significance of changes in the variability of r
MeSH term(s)	Animals ; Humans ; Infant, Newborn ; Animals, Newborn ; Gene Expression ; Hypoxia ; Infant, Premature ; Inflammation/complications ; Intestines ; Oxygen ; Swine ; Random Allocation ; Disease Models, Animal
Chemical Substances	Oxygen (S88TT14065)
Language	English
Publishing date	2023-05-19
Publishing country	United States
Document type	Journal Article
ZDB-ID	4411-8
ISSN	1530-0447 ; 0031-3998
ISSN (online)	1530-0447
ISSN	0031-3998
DOI	10.1038/s41390-023-02657-4
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Article ; Online: Long Non-Coding RNAs in Hypoxia and Oxidative Stress: Novel Insights Investigating a Piglet Model of Perinatal Asphyxia

Grebstad Tune, Benedicte / Melheim, Maria / Åsegg-Atneosen, Monica / Dotinga, Baukje / Saugstad, Ola Didrik / Solberg, Rønnaug / Baumbusch, Lars Oliver

Biology (Basel). 2023 Apr. 04, v. 12, no. 4

2023

Abstract	Birth asphyxia is the leading cause of death and disability in young children worldwide. Long non-coding RNAs (lncRNAs) may provide novel targets and intervention strategies due to their regulatory potential, as demonstrated in various diseases and conditions. We investigated cardinal lncRNAs involved in oxidative stress, hypoxia, apoptosis, and DNA damage using a piglet model of perinatal asphyxia. A total of 42 newborn piglets were randomized into 4 study arms: (1) hypoxia–normoxic reoxygenation, (2) hypoxia–3 min of hyperoxic reoxygenation, (3) hypoxia–30 min of hyperoxic reoxygenation, and (4) sham-operated controls. The expression of lncRNAs BDNF-AS, H19, MALAT1, ANRIL, TUG1, and PANDA, together with the related target genes VEGFA, BDNF, TP53, HIF1α, and TNFα, was assessed in the cortex, the hippocampus, the white matter, and the cerebellum using qPCR and Droplet Digital PCR. Exposure to hypoxia–reoxygenation significantly altered the transcription levels of BDNF-AS, H19, MALAT1, and ANRIL. BDNF-AS levels were significantly enhanced after both hypoxia and subsequent hyperoxic reoxygenation, 8% and 100% O₂, respectively. Our observations suggest an emerging role for lncRNAs as part of the molecular response to hypoxia-induced damages during perinatal asphyxia. A better understanding of the regulatory properties of BDNF-AS and other lncRNAs may reveal novel targets and intervention strategies in the future.
Keywords	DNA damage ; apoptosis ; asphyxia ; cerebellum ; cortex ; death ; droplets ; hippocampus ; hypoxia ; models ; neonates ; oxidative stress ; piglets
Language	English
Dates of publication	2023-0404
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article ; Online
ZDB-ID	2661517-4
ISSN	2079-7737
ISSN	2079-7737
DOI	10.3390/biology12040549
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Assessing nuclear versus mitochondrial cell-free DNA (cfDNA) by qRT-PCR and droplet digital PCR using a piglet model of perinatal asphyxia

Bitenc, Marie / Grebstad Tune, Benedicte / Melheim, Maria / Atneosen-Åsegg, Monica / Lai, Xiaoran / Rajar, Polona / Solberg, Rønnaug / Baumbusch, Lars Oliver

Mol Biol Rep. 2023 Feb., v. 50, no. 2 p.1533-1544

2023

Abstract: BACKGROUND: Since the discovery more than half a century ago, cell-free DNA (cfDNA) has become an attractive objective in multiple diagnostic, prognostic, and monitoring settings. However, despite the increasing number of cfDNA applications in liquid ... ...

Abstract	BACKGROUND: Since the discovery more than half a century ago, cell-free DNA (cfDNA) has become an attractive objective in multiple diagnostic, prognostic, and monitoring settings. However, despite the increasing number of cfDNA applications in liquid biopsies, we still lack a comprehensive understanding of the nature of cfDNA including optimal assessment. In the presented study, we continued testing and validation of common techniques for cfDNA extraction and quantification (qRT-PCR or droplet digital PCR) of nuclear- and mitochondrial cfDNA (ncfDNA and mtcfDNA) in blood, using a piglet model of perinatal asphyxia to determine potential temporal and quantitative changes at the levels of cfDNA. METHODS AND RESULTS: Newborn piglets (n = 19) were either exposed to hypoxia (n = 11) or were part of the sham-operated control group (n = 8). Blood samples were collected at baseline (= start) and at the end of hypoxia or at 40–45 min for the sham-operated control group. Applying the qRT-PCR method, ncfDNA concentrations in piglets exposed to hypoxia revealed an increasing trend from 7.1 ng/ml to 9.5 ng/ml for HK2 (hexokinase 2) and from 4.6 ng/ml to 7.9 ng/ml for β-globulin, respectively, whereas the control animals showed a more balanced profile. Furthermore, median levels of mtcfDNA were much higher in comparison to ncfDNA, but without significant differences between intervention versus the control group. CONCLUSIONS: Both, qRT-PCR and the droplet digital PCR technique identified overall similar patterns for the concentration changes of cfDNA; but, the more sensitive digital PCR methodology might be required to identify minimal responses.
Keywords	DNA ; asphyxia ; blood ; droplets ; hexokinase ; hypoxia ; liquids ; mitochondria ; models ; neonates ; piglets ; polymerase chain reaction
Language	English
Dates of publication	2023-02
Size	p. 1533-1544.
Publishing place	Springer Netherlands
Document type	Article ; Online
ZDB-ID	186544-4
ISSN	1573-4978 ; 0301-4851
ISSN (online)	1573-4978
ISSN	0301-4851
DOI	10.1007/s11033-022-08135-0
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Article ; Online: Splanchnic oxygen saturation during reoxygenation with 21% or 100% O

Dotinga, Baukje M / Solberg, Rønnaug / Saugstad, Ola D / Bos, Arend F / Kooi, Elisabeth M W

Pediatric research

2021 Volume 92, Issue 2, Page(s) 445–452

Abstract: Background: Increasing evidence recognizes the harm of excess oxygen to lungs, eyes, and brain of preterm infants, but not yet to the intestine. We assessed changes in splanchnic oxygenation during reoxygenation with 21% compared to 100% O: Methods: ... ...

Abstract	Background: Increasing evidence recognizes the harm of excess oxygen to lungs, eyes, and brain of preterm infants, but not yet to the intestine. We assessed changes in splanchnic oxygenation during reoxygenation with 21% compared to 100% O Methods: We randomized 25 piglets to control or intervention. Intervention groups underwent global hypoxia until acidosis and hypotension occurred. Piglets were reoxygenated for 30 min with 21% or 100% O Results: Reoxygenation after global hypoxia restored r Conclusion: Reoxygenation after global hypoxia improves splanchnic oxygenation, but is associated with reduced variability of r Impact: Splanchnic oxygenation improves during reoxygenation after global hypoxia, though reoxygenation with 100% O
MeSH term(s)	Animals ; Animals, Newborn ; Humans ; Hyperoxia ; Hypoxia ; Infant, Newborn ; Infant, Premature ; Oxygen ; Oxygen Saturation ; Swine
Chemical Substances	Oxygen (S88TT14065)
Language	English
Publishing date	2021-11-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	4411-8
ISSN	1530-0447 ; 0031-3998
ISSN (online)	1530-0447
ISSN	0031-3998
DOI	10.1038/s41390-021-01819-6
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Article ; Online: Assessing nuclear versus mitochondrial cell-free DNA (cfDNA) by qRT-PCR and droplet digital PCR using a piglet model of perinatal asphyxia.

Bitenc, Marie / Grebstad Tune, Benedicte / Melheim, Maria / Atneosen-Åsegg, Monica / Lai, Xiaoran / Rajar, Polona / Solberg, Rønnaug / Baumbusch, Lars Oliver

Molecular biology reports

2022 Volume 50, Issue 2, Page(s) 1533–1544

Abstract: Background: Since the discovery more than half a century ago, cell-free DNA (cfDNA) has become an attractive objective in multiple diagnostic, prognostic, and monitoring settings. However, despite the increasing number of cfDNA applications in liquid ... ...

Abstract	Background: Since the discovery more than half a century ago, cell-free DNA (cfDNA) has become an attractive objective in multiple diagnostic, prognostic, and monitoring settings. However, despite the increasing number of cfDNA applications in liquid biopsies, we still lack a comprehensive understanding of the nature of cfDNA including optimal assessment. In the presented study, we continued testing and validation of common techniques for cfDNA extraction and quantification (qRT-PCR or droplet digital PCR) of nuclear- and mitochondrial cfDNA (ncfDNA and mtcfDNA) in blood, using a piglet model of perinatal asphyxia to determine potential temporal and quantitative changes at the levels of cfDNA. Methods and results: Newborn piglets (n = 19) were either exposed to hypoxia (n = 11) or were part of the sham-operated control group (n = 8). Blood samples were collected at baseline (= start) and at the end of hypoxia or at 40-45 min for the sham-operated control group. Applying the qRT-PCR method, ncfDNA concentrations in piglets exposed to hypoxia revealed an increasing trend from 7.1 ng/ml to 9.5 ng/ml for HK2 (hexokinase 2) and from 4.6 ng/ml to 7.9 ng/ml for β-globulin, respectively, whereas the control animals showed a more balanced profile. Furthermore, median levels of mtcfDNA were much higher in comparison to ncfDNA, but without significant differences between intervention versus the control group. Conclusions: Both, qRT-PCR and the droplet digital PCR technique identified overall similar patterns for the concentration changes of cfDNA; but, the more sensitive digital PCR methodology might be required to identify minimal responses.
MeSH term(s)	Animals ; Swine ; Cell-Free Nucleic Acids/genetics ; Asphyxia ; Polymerase Chain Reaction/methods ; Liquid Biopsy ; Hypoxia
Chemical Substances	Cell-Free Nucleic Acids
Language	English
Publishing date	2022-12-13
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	186544-4
ISSN	1573-4978 ; 0301-4851
ISSN (online)	1573-4978
ISSN	0301-4851
DOI	10.1007/s11033-022-08135-0
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Article ; Online: A Piglet Perinatal Asphyxia Model to Study Cardiac Injury and Hemodynamics after Cardiac Arrest, Resuscitation, and the Return of Spontaneous Circulation.

Stenersen, Eydis Oddsdottir / Olsen, Annette / Melheim, Maria / Solberg, Rønnaug / Dannevig, Ingrid / Schmölzer, Georg / Cheung, Po-Yin / Nakstad, Britt / Saugstad, Ola Didrik / Rønnestad, Arild / Solevåg, Anne Lee

Journal of visualized experiments : JoVE

2023 , Issue 191

Abstract: Neonatal piglets have been extensively used as translational models for perinatal asphyxia. In 2007, we adapted a well-established piglet asphyxia model by introducing cardiac arrest. This enabled us to study the impact of severe asphyxia on key outcomes, ...

Abstract	Neonatal piglets have been extensively used as translational models for perinatal asphyxia. In 2007, we adapted a well-established piglet asphyxia model by introducing cardiac arrest. This enabled us to study the impact of severe asphyxia on key outcomes, including the time taken for the return of spontaneous circulation (ROSC), as well as the effect of chest compressions according to alternative protocols for cardiopulmonary resuscitation. Due to the anatomical and physiological similarities between piglets and human neonates, piglets serve as good models in studies of cardiopulmonary resuscitation and hemodynamic monitoring. In fact, this cardiac arrest model has provided evidence for guideline development through research on resuscitation protocols, pathophysiology, biomarkers, and novel methods for hemodynamic monitoring. Notably, the incidental finding that a substantial fraction of piglets have pulseless electrical activity (PEA) during cardiac arrest may increase the applicability of the model (i.e., it may be used to study pathophysiology extending beyond the perinatal period). However, the model generation is technically challenging and requires various skill sets, dedicated personnel, and a fine balance of the measures, including the surgical protocols and the use of sedatives/analgesics, to ensure a reasonable rate of survival. In this paper, the protocol is described in detail, as well as experiences with adaptations to the protocol over the years.
MeSH term(s)	Animals ; Swine ; Humans ; Infant, Newborn ; Asphyxia ; Return of Spontaneous Circulation ; Heart Arrest/therapy ; Hemodynamics ; Cardiopulmonary Resuscitation/methods ; Asphyxia Neonatorum/therapy ; Disease Models, Animal
Language	English
Publishing date	2023-01-13
Publishing country	United States
Document type	Journal Article ; Video-Audio Media ; Research Support, Non-U.S. Gov't
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/64788
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Article ; Online: Quantification of circulating cell-free DNA (cfDNA) in urine using a newborn piglet model of asphyxia.

Rajar, Polona / Åsegg-Atneosen, Monica / Saugstad, Ola Didrik / Solberg, Rønnaug / Baumbusch, Lars Oliver

PloS one

2019 Volume 14, Issue 12, Page(s) e0227066

Abstract: Cell free DNA (cfDNA) in plasma has been described as a potential diagnostic indicator for a variety of clinical conditions, including neonatal hypoxia. Neonatal hypoxia or perinatal asphyxia is a severe medical condition caused by a temporary ... ...

Abstract	Cell free DNA (cfDNA) in plasma has been described as a potential diagnostic indicator for a variety of clinical conditions, including neonatal hypoxia. Neonatal hypoxia or perinatal asphyxia is a severe medical condition caused by a temporary interruption in oxygen availability during birth. Previously, we have reported temporal changes of cfDNA detected in blood in a newborn piglet model of perinatal asphyxia. However, cfDNA can also be found in other body liquids, opening for a less invasive diagnostic prospective. The objective of this study was to test and establish a reliable method for the isolation and quantification of cfDNA from urine and to explore changes in the quantities of cfDNA using a newborn piglet model of asphyxia. Animals were exposed to hypoxia-reoxygenation (n = 6), hypoxia-reoxygenation + hypothermia (n = 6) or were part of the sham-operated control group (n = 6) and urine samples (n = 18) were collected at 570 minutes post-intervention. Two alternative applications of cfDNA measurement were tested, an indirect method comprising a centrifugation step together with DNA extraction with magnetic beads versus a direct assessment based on two centrifugation steps. CfDNA concentrations were determined by a fluorescent assay using PicoGreen and by qRT-PCR. Genomic (gDNA) and mitochondrial DNA (mtDNA) cfDNA were determined in parallel, taking into account potential differences in the rates of damages caused by oxidative stress. In contrast to previous publications, our results indicate that the direct method is insufficient. Application of the indirect method obtained with the fluorescence assay revealed mean cfDNA levels (SD) of 1.23 (1.76) ng/ml for the hypoxia samples, 4.47 (6.15) ng/ml for the samples exposed to hypoxia + hypothermia and 2.75 (3.62) ng/ml for the control animals. The mean cfDNA levels in piglets exposed to hypoxia + hypothermia revealed significantly higher cfDNA amounts compared to mean cfDNA levels in the samples purely exposed to hypoxia (p < 0.05); however, no significant difference could be determined when compared to the control group (p = 0.09). Application of the indirect method by qRT-PCR revealed mean cfDNA levels of mtDNA and gDNA at the detection limit of the technique and thus no reliable statistics could be performed between the observed cfDNA levels in the investigated groups. The methodology for detection and monitoring of cfDNA in urine has to be further optimized before it can be applied in a clinical setting in the future.
MeSH term(s)	Animals ; Animals, Newborn ; Asphyxia Neonatorum/diagnosis ; Asphyxia Neonatorum/etiology ; Asphyxia Neonatorum/therapy ; Asphyxia Neonatorum/urine ; Biomarkers/blood ; Biomarkers/urine ; Cell-Free Nucleic Acids/blood ; Cell-Free Nucleic Acids/isolation & purification ; Cell-Free Nucleic Acids/urine ; DNA, Mitochondrial/urine ; Disease Models, Animal ; Feasibility Studies ; Female ; Healthy Volunteers ; Humans ; Hypothermia, Induced/adverse effects ; Hypoxia/complications ; Limit of Detection ; Oxygen/administration & dosage ; Pilot Projects ; Swine
Chemical Substances	Biomarkers ; Cell-Free Nucleic Acids ; DNA, Mitochondrial ; Oxygen (S88TT14065)
Language	English
Publishing date	2019-12-31
Publishing country	United States
Document type	Journal Article
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0227066
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