| 1. |
- Beal, Jacob, et al.
(författare)
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Robust estimation of bacterial cell count from optical density
- 2020
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Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 3:1
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Tidskriftsartikel (refereegranskat)abstract
- Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data.
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| 2. |
- Ahmad, Faiyaz, et al.
(författare)
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Regulation of SERCA2 activity by PDE3A in human myocardium: Phosphorylation-dependent interaction of PDE3A1 with SERCA2.
- 2015
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Ingår i: Journal of Biological Chemistry. - 1083-351X. ; 290:11, s. 6763-6776
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Tidskriftsartikel (refereegranskat)abstract
- PDE3 regulates cAMP-mediated signaling in the heart, and PDE3 inhibitors augment contractility in patients with heart failure. Studies in mice showed that PDE3A, not PDE3B, is the subfamily responsible for these inotropic effects, and that murine PDE3A1 associates with SERCA2, PLB and AKAP18 in a multi-protein signalosome in human SR. Immunohistochemical staining demonstrated that PDE3A co-localizes in Z-bands of human cardiac myocytes with desmin, SERCA2, PLB and AKAP18. In human SR fractions, cAMP increased PLB phosphorylation and SERCA2 activity; this was potentiated by PDE3 inhibition but not by PDE4 inhibition. During gel-filtration chromatography of solubilized SR membranes, PDE3 activity was recovered in distinct HMW and LMW peaks. HMW peaks contained PDE3A1 and PDE3A2, while LMW peaks contained PDE3A1, PDE3A2 and PDE3A3. Western blotting showed that endogenous HMW PDE3A1 was the principal PKA-phosphorylated isoform. Phosphorylation of endogenous PDE3A by rPKAc increased cAMP-hydrolytic activity, correlated with shift of PDE3A from LMW to HMW peaks, and increased co-immumoprecipitation of SERCA2, cav3, PKARII, PP2A and AKAP18 with PDE3A. In experiments with recombinant proteins, phosphorylation of rhPDE3A isoforms by rPKAc increased co-immumoprecipitation with rSERCA2 and rAKAP18. Deletion of the rhPDE3A1/PDE3A2 N-terminus blocked interactions with rSERCA2. Serine-to-alanine substitutions identified S292/S293, a site unique to hPDE3A1, as the principal site regulating its interaction with SERCA2. These results indicate that phosphorylation of hPDE3A1 at a PKA site in its unique N-terminal extension promotes its incorporation into SERCA2/AKAP18 signalosomes, where it regulates a discrete cAMP pool that controls contractility by modulating phosphorylation-dependent protein-protein interactions, PLB phosphorylation and SERCA2 activity.
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| 3. |
- Brault, Emily K., et al.
(författare)
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Trophic position and foraging ecology of Ross, Weddell, and crabeater seals revealed by compound-specific isotope analysis
- 2019
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Ingår i: Marine Ecology Progress Series. - : Inter-Research Science Center. - 0171-8630 .- 1616-1599. ; 611, s. 1-18
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Tidskriftsartikel (refereegranskat)abstract
- © The authors 2019. Ross seals Ommatophoca rossii are one of the least studied marine mammals, with little known about their foraging ecology. Research to date using bulk stable isotope analysis suggests that Ross seals have a trophic position intermediate between that of Weddell Leptonychotes weddellii and crabeater Lobodon carcinophaga seals. However, consumer bulk stable isotope values not only reflect trophic dynamics, but also variations in baseline isotope values, which can be substantial. We used compound-specific isotope analysis of amino acids (CSI-AA) to separate isotopic effects of a shifting baseline versus trophic structure on the foraging ecology of these ecologically important Antarctic pinnipeds. We found that Ross seals forage in an open ocean food web, while crabeater and Weddell seals forage within similar food webs closer to shore. However, isotopic evidence suggests that crabeater seals are likely following sea ice, while Weddell seals target productive areas of the continental shelf of West Antarctica. Our CSI-AA data indicate that Ross seals have a high trophic position equivalent to that of Weddell seals, contrary to prior conclusions from nitrogen isotope results on bulk tissues. CSI-AA indicates that crabeater seals are at a trophic position lower than that of Ross and Weddell seals, consistent with a krill-dominated diet. Our results redefine the view of the trophic dynamics and foraging ecology of the Ross seal, and also highlight the importance of quantifying baseline isotope variations in foraging studies.
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| 4. |
- Carter, Jodi M., et al.
(författare)
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Distinct spatial immune microlandscapes are independently associated with outcomes in triple-negative breast cancer
- 2023
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Ingår i: Nature Communications. - : Springer Nature. - 2041-1723. ; 14
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Tidskriftsartikel (refereegranskat)abstract
- The utility of spatial immunobiomarker quantitation in prognostication and therapeutic prediction is actively being investigated in triple-negative breast cancer (TNBC). Here, with high-plex quantitative digital spatial profiling, we map and quantitate intraepithelial and adjacent stromal tumor immune protein microenvironments in systemic treatment-naive (female only) TNBC to assess the spatial context in immunobiomarker-based prediction of outcome. Immune protein profiles of CD45-rich and CD68-rich stromal microenvironments differ significantly. While they typically mirror adjacent, intraepithelial microenvironments, this is not uniformly true. In two TNBC cohorts, intraepithelial CD40 or HLA-DR enrichment associates with better outcomes, independently of stromal immune protein profiles or stromal TILs and other established prognostic variables. In contrast, intraepithelial or stromal microenvironment enrichment with IDO1 associates with improved survival irrespective of its spatial location. Antigen-presenting and T-cell activation states are inferred from eigenprotein scores. Such scores within the intraepithelial compartment interact with PD-L1 and IDO1 in ways that suggest prognostic and/or therapeutic potential. This characterization of the intrinsic spatial immunobiology of treatment-naive TNBC highlights the importance of spatial microenvironments for biomarker quantitation to resolve intrinsic prognostic and predictive immune features and ultimately inform therapeutic strategies for clinically actionable immune biomarkers.
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| 5. |
- Goetz, Charlotte, et al.
(författare)
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Solar Wind Protons in the Diamagnetic Cavity at Comet 67P/Churyumov-Gerasimenko
- 2023
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Ingår i: Journal of Geophysical Research - Space Physics. - : American Geophysical Union (AGU). - 2169-9380 .- 2169-9402. ; 128:4
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Tidskriftsartikel (refereegranskat)abstract
- The plasma environment at a comet can be divided into different regions with distinct plasma characteristics. Two such regions are the solar wind ion cavity, which refers to the part of the outer coma that does not contain any solar wind ions anymore; and the diamagnetic cavity, which is the region of unmagnetized plasma in the innermost coma. From theory and previous observations, it was thought that under usual circumstances no solar wind ion should be observable near or inside of the diamagnetic cavity. For the first time, we report on five observations that show that protons near solar wind energies can also be found inside the diamagnetic cavity. We characterize these proton signatures, where and when they occur, and discuss possible mechanisms that could lead to protons penetrating the inner coma and traversing the diamagnetic cavity boundary. By understanding these observations, we hope to better understand the interaction region of the comet with the solar wind under nonstandard conditions. The protons detected inside the diamagnetic cavity have directions and energies consistent with protons of solar wind origin. The five events occur only at intermediate gas production rates and low cometocentric distances. Charge transfer reactions, high solar wind dynamic pressure and a neutral gas outburst can be ruled out as causes. We suggest that the anomalous appearance of protons in the diamagnetic cavity is due to a specific solar wind configuration where the solar wind velocity is parallel to the interplanetary magnetic field, thus inhibiting mass-loading and deflection.
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| 6. |
- Goetz, Charlotte, et al.
(författare)
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The plasma environment of comet 67P/Churyumov-Gerasimenko
- 2022
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Ingår i: Space Science Reviews. - : Springer. - 0038-6308 .- 1572-9672. ; 218:8
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Forskningsöversikt (refereegranskat)abstract
- The environment of a comet is a fascinating and unique laboratory to study plasma processes and the formation of structures such as shocks and discontinuities from electron scales to ion scales and above. The European Space Agency's Rosetta mission collected data for more than two years, from the rendezvous with comet 67P/Churyumov-Gerasimenko in August 2014 until the final touch-down of the spacecraft end of September 2016. This escort phase spanned a large arc of the comet's orbit around the Sun, including its perihelion and corresponding to heliocentric distances between 3.8 AU and 1.24 AU. The length of the active mission together with this span in heliocentric and cometocentric distances make the Rosetta data set unique and much richer than sets obtained with previous cometary probes. Here, we review the results from the Rosetta mission that pertain to the plasma environment. We detail all known sources and losses of the plasma and typical processes within it. The findings from in-situ plasma measurements are complemented by remote observations of emissions from the plasma. Overviews of the methods and instruments used in the study are given as well as a short review of the Rosetta mission. The long duration of the Rosetta mission provides the opportunity to better understand how the importance of these processes changes depending on parameters like the outgassing rate and the solar wind conditions. We discuss how the shape and existence of large scale structures depend on these parameters and how the plasma within different regions of the plasma environment can be characterised. We end with a non-exhaustive list of still open questions, as well as suggestions on how to answer them in the future.
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| 7. |
- Goetz, Charlotte, et al.
(författare)
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Warm protons at comet 67P/Churyumov-Gerasimenko-implications for the infant bow shock
- 2021
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Ingår i: Annales Geophysicae. - : European Geosciences Union (EGU). - 0992-7689 .- 1432-0576. ; 39:3, s. 379-396
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Tidskriftsartikel (refereegranskat)abstract
- The plasma around comet 67P/Churyumov-Gerasimenko showed remarkable variability throughout the entire Rosetta mission. Plasma boundaries such as the diamagnetic cavity, solar wind ion cavity and infant bow shock separate regions with distinct plasma parameters from each other. Here, we focus on a particular feature in the plasma: warm, slow solar wind protons. We investigate this particular proton population further by focusing on the proton behaviour and surveying all of the Rosetta comet phase data. We find over 300 events where Rosetta transited from a region with fast, cold protons into a region with warm, slow protons. We investigate the properties of the plasma and magnetic field at this boundary and the location where it can be found. We find that the protons are preferentially detected at intermediate gas production rates with a slight trend towards larger cometocentric distances for higher gas production rates. The events can mostly be found in the positive convective electric field hemisphere. These results agree well with simulations of the infant bow shock (IBS), an asymmetric structure in the plasma environment previously detected on only 2 d during the comet phase. The properties of the plasma on both sides of this structure are harder to constrain, but there is a trend towards higher electron flux, lower magnetic field, higher magnetic field power spectral density and higher density in the region that contains the warm protons. This is in partial agreement with the previous IBS definitions; however, it also indicates that the plasma and this structure are highly non-stationary. For future research, Comet Interceptor, with its multi-point measurements, can help to disentangle the spatial and temporal effects and give more clarity on the influence of changing upstream conditions on the movement of boundaries in this unusual environment.
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| 8. |
- Gonzalez-Ericsson, Paula, et al.
(författare)
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The path to a better biomarker: application of a risk management framework for the implementation of PD‐L1 and TILs as immuno‐oncology biomarkers into breast cancer clinical trials and daily practice
- 2020
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Ingår i: Journal of Pathology. - : Wiley. - 1096-9896 .- 0022-3417. ; 250:5, s. 667-684
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Forskningsöversikt (refereegranskat)abstract
- Immune checkpoint inhibitor therapies targeting PD‐1/PD‐L1 are now the standard of care in oncology across several hematologic and solid tumor types, including triple negative breast cancer (TNBC). Patients with metastatic or locally advanced TNBC with PD‐L1 expression on immune cells occupying ≥1% of tumor area demonstrated survival benefit with the addition of atezolizumab to nab‐paclitaxel. However, concerns regarding variability between immunohistochemical PD‐L1 assay performance and inter‐reader reproducibility have been raised. High tumor‐infiltrating lymphocytes (TILs) have also been associated with response to PD‐1/PD‐L1 inhibitors in patients with breast cancer (BC). TILs can be easily assessed on hematoxylin and eosin–stained slides and have shown reliable inter‐reader reproducibility. As an established prognostic factor in early stage TNBC, TILs are soon anticipated to be reported in daily practice in many pathology laboratories worldwide. Because TILs and PD‐L1 are parts of an immunological spectrum in BC, we propose the systematic implementation of combined PD‐L1 and TIL analyses as a more comprehensive immuno‐oncological biomarker for patient selection for PD‐1/PD‐L1 inhibition‐based therapy in patients with BC. Although practical and regulatory considerations differ by jurisdiction, the pathology community has the responsibility to patients to implement assays that lead to optimal patient selection. We propose herewith a risk‐management framework that may help mitigate the risks of suboptimal patient selection for immuno‐therapeutic approaches in clinical trials and daily practice based on combined TILs/PD‐L1 assessment in BC.
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| 9. |
- Jones, Geraint H., et al.
(författare)
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The Comet Interceptor Mission
- 2024
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Ingår i: Space Science Reviews. - : Springer Nature. - 0038-6308 .- 1572-9672. ; 220:1
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Tidskriftsartikel (refereegranskat)abstract
- Here we describe the novel, multi-point Comet Interceptor mission. It is dedicated to the exploration of a little-processed long-period comet, possibly entering the inner Solar System for the first time, or to encounter an interstellar object originating at another star. The objectives of the mission are to address the following questions: What are the surface composition, shape, morphology, and structure of the target object? What is the composition of the gas and dust in the coma, its connection to the nucleus, and the nature of its interaction with the solar wind? The mission was proposed to the European Space Agency in 2018, and formally adopted by the agency in June 2022, for launch in 2029 together with the Ariel mission. Comet Interceptor will take advantage of the opportunity presented by ESA’s F-Class call for fast, flexible, low-cost missions to which it was proposed. The call required a launch to a halo orbit around the Sun-Earth L2 point. The mission can take advantage of this placement to wait for the discovery of a suitable comet reachable with its minimum Δ V capability of 600 ms − 1 . Comet Interceptor will be unique in encountering and studying, at a nominal closest approach distance of 1000 km, a comet that represents a near-pristine sample of material from the formation of the Solar System. It will also add a capability that no previous cometary mission has had, which is to deploy two sub-probes – B1, provided by the Japanese space agency, JAXA, and B2 – that will follow different trajectories through the coma. While the main probe passes at a nominal 1000 km distance, probes B1 and B2 will follow different chords through the coma at distances of 850 km and 400 km, respectively. The result will be unique, simultaneous, spatially resolved information of the 3-dimensional properties of the target comet and its interaction with the space environment. We present the mission’s science background leading to these objectives, as well as an overview of the scientific instruments, mission design, and schedule.
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| 10. |
- Leon-Ferre, Roberto A., et al.
(författare)
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Automated mitotic spindle hotspot counts are highly associated with clinical outcomes in systemically untreated early-stage triple-negative breast cancer
- 2024
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Ingår i: npj Breast Cancer. - : NATURE PORTFOLIO. - 2374-4677. ; 10:1
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Tidskriftsartikel (refereegranskat)abstract
- Operable triple-negative breast cancer (TNBC) has a higher risk of recurrence and death compared to other subtypes. Tumor size and nodal status are the primary clinical factors used to guide systemic treatment, while biomarkers of proliferation have not demonstrated value. Recent studies suggest that subsets of TNBC have a favorable prognosis, even without systemic therapy. We evaluated the association of fully automated mitotic spindle hotspot (AMSH) counts with recurrence-free (RFS) and overall survival (OS) in two separate cohorts of patients with early-stage TNBC who did not receive systemic therapy. AMSH counts were obtained from areas with the highest mitotic density in digitized whole slide images processed with a convolutional neural network trained to detect mitoses. In 140 patients from the Mayo Clinic TNBC cohort, AMSH counts were significantly associated with RFS and OS in a multivariable model controlling for nodal status, tumor size, and tumor-infiltrating lymphocytes (TILs) (p < 0.0001). For every 10-point increase in AMSH counts, there was a 16% increase in the risk of an RFS event (HR 1.16, 95% CI 1.08-1.25), and a 7% increase in the risk of death (HR 1.07, 95% CI 1.00-1.14). We corroborated these findings in a separate cohort of systemically untreated TNBC patients from Radboud UMC in the Netherlands. Our findings suggest that AMSH counts offer valuable prognostic information in patients with early-stage TNBC who did not receive systemic therapy, independent of tumor size, nodal status, and TILs. If further validated, AMSH counts could help inform future systemic therapy de-escalation strategies.
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| 11. |
- Pfeiler, Georg, et al.
(författare)
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Impact of BMI in Patients With Early Hormone Receptor-Positive Breast Cancer Receiving Endocrine Therapy With or Without Palbociclib in the PALLAS Trial
- 2023
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Ingår i: JOURNAL OF CLINICAL ONCOLOGY. - 0732-183X .- 1527-7755. ; 41:33
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Tidskriftsartikel (refereegranskat)abstract
- PURPOSEBMI affects breast cancer risk and prognosis. In contrast to cytotoxic chemotherapy, CDK4/6 inhibitors are given at a fixed dose, irrespective of BMI or weight. This preplanned analysis of the global randomized PALLAS trial investigates the impact of BMI on the side-effect profile, treatment adherence, and efficacy of palbociclib.METHODSPatients were categorized at baseline according to WHO BMI categories. Neutropenia rates were assessed with univariable and multivariable logistic regression. Time to early discontinuation of palbociclib was analyzed with Fine and Gray competing risk models. Unstratified Cox models were used to investigate the association between BMI category and time to invasive disease-free survival (iDFS). 95% CIs were derived.RESULTSOf 5,698 patients included in this analysis, 68 (1.2%) were underweight, 2,082 (36.5%) normal weight, 1,818 (31.9%) overweight, and 1,730 (30.4%) obese at baseline. In the palbociclib arm, higher BMI was associated with a significant decrease in neutropenia (unadjusted odds ratio for 1-unit change, 0.93; 95% CI, 0.91 to 0.94; adjusted for age, race ethnicity, region, chemotherapy use, and Eastern Cooperative Oncology Group at baseline, 0.93; 95% CI, 0.92 to 0.95). This translated into a significant decrease in treatment discontinuation rate with higher BMI (adjusted hazard ratio [HR] for 10-unit change, 0.75; 95% CI, 0.67 to 0.83). There was no significant improvement in iDFS with the addition of palbociclib to ET in any weight category (normal weight HR, 0.84; 95% CI, 0.63 to 1.12; overweight HR, 1.10; 95% CI, 0.82 to 1.49; and obese HR, 0.95; 95% CI, 0.69 to 1.30) in this analysis early in follow-up (31 months).CONCLUSIONThis preplanned analysis of the PALLAS trial demonstrates a significant impact of BMI on side effects, dose reductions, early treatment discontinuation, and relative dose intensity. Additional long-term follow-up will further evaluate whether BMI ultimately affects outcome. This preplanned analysis of the PALLAS trial demonstrates a significant impact of BMI on side effects, dose reductions, early treatment discontinuation, and relative dose intensity.
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| 12. |
- Watts, Jennifer D., et al.
(författare)
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Carbon uptake in Eurasian boreal forests dominates the high-latitude net ecosystem carbon budget
- 2023
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Ingår i: Global Change Biology. - : Wiley. - 1354-1013 .- 1365-2486. ; 29:7, s. 1870-1889
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Tidskriftsartikel (refereegranskat)abstract
- Arctic-boreal landscapes are experiencing profound warming, along with changes in ecosystem moisture status and disturbance from fire. This region is of global importance in terms of carbon feedbacks to climate, yet the sign (sink or source) and magnitude of the Arctic-boreal carbon budget within recent years remains highly uncertain. Here, we provide new estimates of recent (2003–2015) vegetation gross primary productivity (GPP), ecosystem respiration (Reco), net ecosystem CO2 exchange (NEE; Reco − GPP), and terrestrial methane (CH4) emissions for the Arctic-boreal zone using a satellite data-driven process-model for northern ecosystems (TCFM-Arctic), calibrated and evaluated using measurements from >60 tower eddy covariance (EC) sites. We used TCFM-Arctic to obtain daily 1-km2 flux estimates and annual carbon budgets for the pan-Arctic-boreal region. Across the domain, the model indicated an overall average NEE sink of −850 Tg CO2-C year−1. Eurasian boreal zones, especially those in Siberia, contributed to a majority of the net sink. In contrast, the tundra biome was relatively carbon neutral (ranging from small sink to source). Regional CH4 emissions from tundra and boreal wetlands (not accounting for aquatic CH4) were estimated at 35 Tg CH4-C year−1. Accounting for additional emissions from open water aquatic bodies and from fire, using available estimates from the literature, reduced the total regional NEE sink by 21% and shifted many far northern tundra landscapes, and some boreal forests, to a net carbon source. This assessment, based on in situ observations and models, improves our understanding of the high-latitude carbon status and also indicates a continued need for integrated site-to-regional assessments to monitor the vulnerability of these ecosystems to climate change.
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