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Mesenchymal stromal cells (MSC) hold promise for both cell replacement and immune modulation strategies owing to their progenitor and non-progenitor functions, respectively. Characterization of MSC from different sources is an important and necessary step before clinical use of these cells is widely adopted. Little is known about the biology and function of canine MSC compared to their mouse or human counterparts. This knowledge-gap impedes development of canine evidence-based MSC technologies. We hypothesized that canine adipose tissue (AT) and bone marrow (BM) MSC (derived from the same dogs) will have similar differentiation and immune modulatory profiles. Our objectives were to evaluate progenitor and non-progenitor functions as well as other characteristics of AT- and BM-MSC including 1) proliferation rate, 2) cell surface marker expression, 3) DNA methylation levels, 4) potential for trilineage differentiation towards osteogenic, adipogenic, and chondrogenic cell fates, and 5) immunomodulatory potency in vitro. 1) AT-MSC proliferated at more than double the rate of BM-MSC (population doubling times in days) for passage (P) 2, AT: 1.69, BM: 3.81; P3, AT: 1.80, BM: 4.06; P4, AT: 2.37, BM: 5.34; P5, AT: 3.20, BM: 7.21). 2) Canine MSC, regardless of source, strongly expressed cell surface markers MHC I, CD29, CD44, and CD90, and were negative for MHC II and CD45. They also showed moderate expression of CD8 and CD73 and mild expression of CD14. Minor differences were found in expression of CD4 and CD34. 3) Global DNA methylation levels were significantly lower in BM-MSC compared to AT-MSC. 4) Little difference was found between AT- and BM-MSC in their potential for adipogenesis and osteogenesis. Chondrogenesis was poor to absent for both sources in spite of adding varying levels of bone-morphogenic protein to our standard transforming growth factor (TGF-β3)-based induction medium. 5) Immunomodulatory capacity was equal regardless of cell source when tested in mitogen-stimulated lymphocyte reactions. Priming of MSC with pro-inflammatory factors interferon-gamma and/or tumour necrosis factor did not increase the lymphocyte suppressive properties of the MSC compared to untreated MSC. No significant differences were found between AT- and BM-MSC with regard to their immunophenotype, progenitor, and non-progenitor functions. Both MSC populations showed strong adipogenic and osteogenic potential and poor chondrogenic potential. Both significantly suppressed stimulated peripheral blood mononuclear cells. The most significant differences found were the higher isolation success and proliferation rate of AT-MSC, which could be realized as notable benefits of their use over BM-MSC.

This paper analyzes effects of the mutual presentation of weak and strong arguments. Departing from the prevalent “the-more-the-better” heuristic, our research scrutinizes whether the inclusion of weak arguments enhances or diminishes the persuasive impact of strong arguments. Leveraging insights from judgment formation literature, we conducted four experimental studies on political and health-related topics to unravel whether the presenting weak arguments strengthens the persuasive effect of a strong argument (adding) or actually weakens this persuasive effect (averaging). The results show that providing supporting arguments of moderate strength along with a strong argument increases persuasion, representing an additive pattern. However, presenting weak supporting arguments along with a strong argument reduces the persuasive effect of the strong argument, representing an averaging pattern. Exposure to weak arguments diminishes the strength of strong ones, suggesting the omission of weak arguments. These findings underscore the vital role of strategically selecting arguments to optimize persuasion across disciplines.

Mesenchymal stromal cells (MSC) are increasingly investigated for their clinical utility in dogs. Fetal bovine serum (FBS) is a common culture supplement used for canine MSC expansion. However, FBS content is variable, its clinical use carries risk of an immune response, and its cost is increasing due to global demand. Platelet lysate (PL) has proven to be a suitable alternative to FBS for expansion of human MSC. We hypothesized that canine adipose tissue (AT) and bone marrow (BM) MSC could be isolated and expanded equally in PL and FBS at conventionally-used concentrations with differentiation of these MSC unaffected by choice of supplement. Our objectives were to evaluate the use of canine PL in comparison with FBS at four stages: 1) isolation, 2) proliferation, 3) spontaneous differentiation, and 4) directed differentiation. 1) Medium with 10% PL was unable to isolate MSC. 2) MSC, initially isolated in FBS-supplemented media, followed a dose-dependent response with no significant difference between PL and FBS cultures at up to 20% (AT) or 30% (BM) enrichment. Beyond these respective peaks, proliferation fell in PL cultures only, while a continued dose-dependent proliferation response was noted in FBS cultures. 3) Further investigation indicated PL expansion culture was inducing spontaneous adipogenesis in concentrations as low as 10% and as early as 4 days in culture. 4) MSC isolated in FBS, but expanded in either FBS or PL, maintained ability to undergo directed adipogenesis and osteogenesis, but not chondrogenesis. Canine PL did not support establishment of MSC colonies from AT and BM, nor expansion of MSC, which appear to undergo spontaneous adipogenesis in response to PL exposure. In vivo studies are warranted to determine if concurrent use of MSC with any platelet-derived products such as platelet-rich plasma are associated with synergistic, neutral or antagonistic effects.

Background Nervous systems originated before the split of Proto- and Deuterostomia, more than 600 million years ago. Four animal phyla (Cnidaria, Placozoa, Ctenophora, Porifera) diverged before this split and studying these phyla could give us important information on the evolution of the nervous system. Here, we have annotated the neuropeptide preprohormone genes of twenty species belonging to the subclass Hexacorallia or Ceriantharia (Anthozoa: Cnidaria), using thirty-seven publicly accessible genome or transcriptome databases. Studying hexacorals is important, because they are versatile laboratory models for development (e.g., Nematostella vectensis ) and symbiosis (e.g., Exaiptasia diaphana ) and also are prominent reef-builders. Results We found that each hexacoral or ceriantharian species contains five to ten neuropeptide preprohormone genes. Many of these preprohormones contain multiple copies of immature neuropeptides, which can be up to 50 copies of identical or similar neuropeptide sequences. We also discovered preprohormones that only contained one neuropeptide sequence positioned directly after the signal sequence. Examples of them are neuropeptides that terminate with the sequence RWamide (the Antho-RWamides). Most neuropeptide sequences are N-terminally protected by pyroglutamyl (pQ) or one or more prolyl residues, while they are C-terminally protected by an amide group. Previously, we isolated and sequenced small neuropeptides from hexacorals that were N-terminally protected by an unusual L-3-phenyllactyl group. In our current analysis, we found that these N-phenyllactyl-peptides are derived from N-phenylalanyl-peptides located directly after the signal sequence of the preprohormone. The N-phenyllactyl- peptides appear to be confined to the hexacorallian order Actiniaria and do not occur in other cnidarians. On the other hand, (1) the neuropeptide Antho-RFamide (pQGRFamide); (2) peptides with the C-terminal sequence GLWamide; and (3) tetrapeptides with the X 1 PRX 2 amide consensus sequence (most frequently GPRGamide) are ubiquitous in Hexacorallia. Conclusions We found GRFamide, GLWamide, and X 1 PRX 2 amide peptides in all tested Hexacorallia. Previously, we discovered these three neuropeptide classes also in Cubozoa, Scyphozoa, and Staurozoa, indicating that these neuropeptides originated in the common cnidarian ancestor and are evolutionarily ancient. In addition to these ubiquitous neuropeptides, other neuropeptides appear to be confined to specific cnidarian orders or subclasses.

We present an analysis of atmospheric transport impact on estimating CO2 fluxes using two atmospheric inversion systems (CarboScope-Regional (CSR) and Lund University Modular Inversion Algorithm (LUMIA)) over Europe in 2018. The main focus of this study is to quantify the dominant drivers of spread amid CO2 estimates derived from atmospheric tracer inversions. The Lagrangian transport models STILT (Stochastic Time-Inverted Lagrangian Transport) and FLEXPART (FLEXible PARTicle) were used to assess the impact of mesoscale transport. The impact of lateral boundary conditions for CO2 was assessed by using two different estimates from the global inversion systems CarboScope (TM3) and TM5-4DVAR. CO2 estimates calculated with an ensemble of eight inversions differing in the regional and global transport models, as well as the inversion systems, show a relatively large spread for the annual fluxes, ranging between -0.72 and 0.20 PgC yr-1, which is larger than the a priori uncertainty of 0.47 PgC yr-1. The discrepancies in annual budget are primarily caused by differences in the mesoscale transport model (0.51 PgC yr-1), in comparison with 0.23 and 0.10 PgC yr-1 that resulted from the far-field contributions and the inversion systems, respectively. Additionally, varying the mesoscale transport caused large discrepancies in spatial and temporal patterns, while changing the lateral boundary conditions led to more homogeneous spatial and temporal impact. We further investigated the origin of the discrepancies between transport models. The meteorological forcing parameters (forecasts versus reanalysis obtained from ECMWF data products) used to drive the transport models are responsible for a small part of the differences in CO2 estimates, but the largest impact seems to come from the transport model schemes. Although a good convergence in the differences between the inversion systems was achieved by applying a strict protocol of using identical prior fluxes and atmospheric datasets, there was a non-negligible impact arising from applying a different inversion system. Specifically, the choice of prior error structure accounted for a large part of system-to-system differences.

Multipotent mesenchymal stromal cells (MSC) have attracted interest for their cytotherapeutic potential, partly due to their immunomodulatory abilities. The aim of this study was to test the robustness of our equine cord blood (CB) MSC isolation protocol, to characterize the CB-MSC before and after cryopreservation, and to evaluate their immunosuppressive phenotype. We hypothesized that MSC can be consistently isolated from equine CB, have unique and reproducible marker expression and in vitro suppress lymphoproliferation. Preliminary investigation of constitutive cytoplasmic Toll-like receptor (TLR) 3 and 4 expression was also preformed due to their possible association with anti- or pro-inflammatory MSC phenotypes, respectively. Surface markers were assessed for antigen and mRNA expression by flow cytometry and quantitative polymerase chain reaction (qPCR). Immunomodulatory properties were evaluated in mixed lymphocyte reaction assays, and TLR3 and TLR4 expression were measured by qPCR and immunocytochemistry (ICC). CB-MSC were isolated from each off nine cord blood samples. CB-MSC highly expressed CD29, CD44, CD90, and lacked or had low expression of major histocompatibility complex (MHC) class I, MHC-II, CD4, CD8, CD11a/18 and CD73 before and after cryopreservation. CB-MSC suppressed in vitro lymphoproliferation and constitutively expressed TLR4. Our findings confirmed CB as a reliable MSC source, provides an association of surface marker phenotype and mRNA expression and suggest anti-inflammatory properties of CB-MSC. The relationship between TLRs and lymphocyte function warrants further investigation.

Atmospheric inversions have been used for the past two decades to derive large-scale constraints on the sources and sinks of CO2 into the atmosphere. The development of dense in situ surface observation networks, such as ICOS in Europe, enables in theory inversions at a resolution close to the country scale in Europe. This has led to the development of many regional inversion systems capable of assimilating these high-resolution data, in Europe and elsewhere. The EUROCOM (European atmospheric transport inversion comparison) project is a collaboration between seven European research institutes, which aims at producing a collective assessment of the net carbon flux between the terrestrial ecosystems and the atmosphere in Europe for the period 2006–2015. It aims in particular at investigating the capacity of the inversions to deliver consistent flux estimates from the country scale up to the continental scale. The project participants were provided with a common database of in situ-observed CO2 concentrations (including the observation sites that are now part of the ICOS network) and were tasked with providing their best estimate of the net terrestrial carbon flux for that period, and for a large domain covering the entire European Union. The inversion systems differ by the transport model, the inversion approach, and the choice of observation and prior constraints, enabling us to widely explore the space of uncertainties. This paper describes the intercomparison protocol and the participating systems, and it presents the first results from a reference set of inversions, at the continental scale and in four large regions. At the continental scale, the regional inversions support the assumption that European ecosystems are a relatively small sink (-0.21±0.2 Pg C yr−1). We find that the convergence of the regional inversions at this scale is not better than that obtained in state-of-the-art global inversions. However, more robust results are obtained for sub-regions within Europe, and in these areas with dense observational coverage, the objective of delivering robust country-scale flux estimates appears achievable in the near future.

Ignoring the diurnal cycle in surface-to-atmosphere CO2 fluxes leads to a systematic bias in CO2 mole fraction simulations sampled at daytime because the daily mean flux systematically misses the CO2 uptake during the daytime hours. In an atmospheric inversion using daytime-selected CO2 measurements at most continental sites and not resolving diurnal cycles in the flux, this leads to systematic biases in the estimates of the annual sources and sinks of atmospheric CO2. This study focuses on quantifying the impact of this diurnal cycle effect on the annual carbon fluxes estimated with the CarboScope (CS) atmospheric inversion at regional, continental, and global scales for the period of time 2010–2020. Our analysis is based on biogenic fluxes of hourly net ecosystem exchange (NEE) obtained from the data-driven FLUXCOM-X estimates, together with global and regional atmospheric transport models. Differences between CO2 mixing ratios simulated with daily averaged and hourly NEE from FLUXCOM-X range between around -2.5 and 7 ppm averaged annually throughout a site network across the world. These differences lead to systematic biases in CO2 flux estimates from the atmospheric inversions. Although the impact on the global total flux is negligible (around 2 % of the overall land flux of -1.79 Pg C yr−1), we find significant biases in the annual flux budgets at continental and regional scales. For Europe, the annual mean difference in the fluxes arising indirectly from the diurnal cycle of CO2 through the boundary condition amounts to around 48 % of the annual posterior fluxes (0.31 Pg C yr−1) estimated with CarboScope-Regional (CSR). Furthermore, the differences in NEE estimates calculated with CS increase the magnitude of the flux budgets for some regions such as North American temperate forests and northern Africa by a factor of about 1.5. To the extent that FLUXCOM-X diurnal cycles are realistic at all latitudes and for the station set including many continental stations as used in our inversions here, we conclude that ignoring the diurnal variations in the land CO2 flux leads to overestimation of both CO2 sources in the tropical lands and CO2 sinks in the temperate zones.

Three-hourly net ecosystem exchange (NEE) is estimated at spatial scales of 0.25∘ over the European continent, based on the pre-operational inverse modelling framework “CarboScope Regional” (CSR) for the years 2006 to 2019. To assess the uncertainty originating from the choice of a priori flux models and observational data, ensembles of inversions were produced using three terrestrial ecosystem flux models, two ocean flux models, and three sets of atmospheric stations. We find that the station set ensemble accounts for 61 % of the total spread of the annually aggregated fluxes over the full domain when varying all these elements, while the biosphere and ocean ensembles resulted in much smaller contributions to the spread of 28 % and 11 %, respectively. These percentages differ over the specific regions of Europe, based on the availability of atmospheric data. For example, the spread of the biosphere ensemble is prone to be larger in regions that are less constrained by CO2 measurements. We investigate the impact of unprecedented increase in temperature and simultaneous reduction in soil water content (SWC) observed in 2018 and 2019 on the carbon cycle. We find that NEE estimates during these 2 years suggest an impact of drought occurrences represented by the reduction in net primary productivity (NPP), which in turn leads to less CO2 uptake across Europe in 2018 and 2019, resulting in anomalies of up to 0.13 and 0.07 PgC yr−1 above the climatological mean, respectively. Annual temperature anomalies also exceeded the climatological mean by 0.46 ∘C in 2018 and by 0.56 ∘C in 2019, while Standardised Precipitation–Evaporation Index (SPEI) anomalies declined to −0.20 and −0.05 SPEI units below the climatological mean in both 2018 and 2019, respectively. Therefore, the biogenic fluxes showed a weaker sink of CO2 in both 2018 and 2019 (−0.22 ± 0.05 and −0.28 ± 0.06 PgC yr−1, respectively) in comparison with the mean −0.36 ± 0.07 PgC yr−1 calculated over the full analysed period (i.e. 14 years). These translate into a continental-wide reduction in the annual sink by 39 % and 22 %, respectively, larger than the typical year-to-year standard deviation of 19 % observed over the full period.

Multi-material 3D printing concerns the use of two or more 3D printable materials within a single printed part. The result is a composite that benefits from the combined properties of the individual 3D printed materials. Typically, a distinct differentiation between material properties can only be achieved using multiple feedstocks and advanced engineering solutions. In this work, we create multi-material 3D printed photopolymer parts from a single monomer mixture through simple adjustments in printing temperature and light intensity. We achieve this by employing a liquid crystalline (LC) monomer that forms a highly stable LC phase in conjunction with a trifunctional thiol crosslinker. A drastic change in mechanical and optical properties was achieved depending on the presence of an LC phase during polymerization. The proof of principle from bulk experiments could be translated fully into 3D printing, achieving pixel-to-pixel resolution of the material properties solely guided by changing the printing parameters temperature and light intensity. The versatility of produced multi-material composite parts is demonstrated in shape memory applications and methods for chemical data storage and encryption. 3D printing can be used to manufacture composite materials, but to modify material properties multiple feedstocks are required. Here the authors modify printing temperature or light intensity to modify the material properties and achieve property differentiation from a single feedstock.