Akt-dependent Pp2a activity is required for epidermal barrier formation during late embryonic development

Acquisition of epidermal barrier function occurs late in mouse gestation. Several days before birth a wave of barrier acquisition sweeps across murine fetal skin, converging on dorsal and ventral midlines. We investigated the molecular pathways active during epidermal barrier formation. Akt signaling increased as the barrier wave crossed epidermis and Jun was transiently dephosphorylated. Inhibitor experiments on embryonic explants showed that the dephosphorylation of Jun was dependent on both Akt and protein phosphatase 2A (Pp2a). Inhibition of Pp2a and Akt signaling also caused defects in epidermal barrier formation. These data are compatible with a model for developmental barrier acquisition mediated by Pp2a regulation of Jun dephosphorylation, downstream of Akt signaling. Support for this model was provided by siRNA-mediated knockdown of Ppp2r2a (Pr55α or B55α), a regulatory subunit of Pp2a expressed in an Akt-dependent manner in epidermis during barrier formation. Ppp2r2a reduction caused significant increase in Jun phosphorylation and interfered with the acquisition of barrier function, with barrier acquisition being restored by inhibition of Jun phosphorylation. Our data provide strong evidence that Ppp2r2a is a regulatory subunit of Pp2a that targets this phosphatase to Jun, and that Pp2a action is necessary for barrier formation. We therefore describe a novel Akt-dependent Pp2a activity that acts at least partly through Jun to affect initial barrier formation during late embryonic epidermal development

3D co-cultures of osteoblasts and endothelial cells in DegraPol foam: Histological and high field MRI analyses of pre-engineered capillary networks in bone grafts

Tissue engineering of bone grafts was addressed in a critical size model on the chick chorioallantoic membrane model (CAM assay), using DegraPol(R) (DP) foam as scaffold material. The scaffolds were seeded with cultures of human osteoblasts (OB) and human en notdo notthelial cells (EC), respectively, or with a co-culture of the two cell types (control: no cells). In vitro samples (7 days cultivation) and ex vivo CAM samples at incubation day 15 (ID 15) were analyzed by high field magnetic resonance imaging (MRI) and histology. The co-culture system performed best with respect to perfusion, as assessed by contrast-enhanced MRI using Gd-DTPA. The scaffold seeded by the co-culture supported an increased vascular ingrowth, which was confirmed by histological analysis. DP foam is a suitable scaffold for bone tissue engineering and the MRI technique allows for non-destructive and quantitative assessment of perfusion capability during early stages of bone forming constructs

Studies of insect temporal trends must account for the complex sampling histories inherent to many long-term monitoring efforts

Crossley et al. (2020)1 examine patterns of change in insect abundance and diversity across US Long-Term Ecological Research (LTER) sites, concluding “a lack of overall increase or decline”. This is notable if true, given mixed conclusions in the literature regarding the nature and ubiquity of insect declines across regions and insect taxonomic groups2–6. The data analyzed, downloaded from and collected by US LTER sites, represent unique time series of arthropod abundances. These long-term datasets often provide critical insights, capturing both steady changes and responses to sudden unpredictable events. However, a number of the included datasets are not suitable for estimating long-term observational trends because they come from experiments or have methodological inconsistencies. Additionally, long-term ecological datasets are rarely uniform in sampling effort across their full duration as a result of the changing goals and abilities of a research site to collect data7. We suggest that Crossley et al.’s results rely upon a key, but flawed, assumption, that sampling was collected “in a consistent way over time within each dataset”. We document problems with data use prior to statistical analyses from eight LTER sites due to datasets not being suitable for long-term trend estimation and not accounting for sampling variation, using the Konza Prairie (KNZ) grasshopper dataset (CGR022) as an example

Laboratory studies of immersion and deposition mode ice nucleation of ozone aged mineral dust particles

Ice nucleation in the atmosphere is central to the understanding the microphysical properties of mixed-phase and cirrus clouds. Ambient conditions such as temperature (T) and relative humidity (RH), as well as aerosol properties such as chemical composition and mixing state play an important role in predicting ice formation in the troposphere. Previous field studies have reported the absence of sulfate and organic compounds on mineral dust ice crystal residuals sampled at mountain top stations or aircraft based measurements despite the long-range transport mineral dust is subjected to. We present laboratory studies of ice nucleation for immersion and deposition mode on ozone aged mineral dust particles for 233 T ns) are reported and observed to increase as a function of decreasing temperature. We present first results that demonstrate enhancement of the ice nucleation ability of aged mineral dust particles in both the deposition and immersion mode due to ageing. We also present the first results to show a suppression of heterogeneous ice nucleation activity without the condensation of a coating of (in)organic material. In immersion mode, low ozone exposed Ka particles showed enhanced ice activity requiring a median freezing temperature of 1.5 K warmer than that of untreated Ka, whereas high ozone exposed ATD particles showed suppressed ice nucleation requiring a median freezing temperature of 3 K colder than that of untreated ATD. In deposition mode, low exposure Ka had ice active fractions of an order of magnitude higher than untreated Ka, whereas high ozone exposed ATD had ice active fractions up to a factor of 4 lower than untreated ATD. From our results, we derive and present parameterizations in terms of ns(T) that can be used in models to predict ice nuclei concentrations based on available aerosol surface area

Exploring the Potential Effect of Phospholipase A2 Antibody to Extend Beef Shelf-Life in a Beef Liposome Model System

Objective: The objective of this study was to utilize a beef liposome model system to investigate if phospholipase-A2 antibody (aPLA2) can be used to inhibit phospholipase-A2 (PLA2) activity to potentially improve beef shelf-life. Study Description: Phospholipids (PL) from 10 U.S. Department of Agriculture choice beef striploin steaks were extracted and split into six treatments: PL (25 mg of PL); aPLA10 (PL + 25 µg of aPLA2); aPLA20 (PL + 50 µg of aPLA2); PLA2 (PL + 10 µg of PLA2); PLA2+aPLA10 (PL + PLA2 + aPLA10); and PLA2+aPLA20 (PL + PLA2 + aPLA20). The model system was under retail display at 39°F and 2300 lux for 7 days. At day 0, aliquots were taken for PL profiling and product ion analysis. At days 0, 1, 4, and 7, aliquots were taken for lipid oxidation analysis. Results: At day 7 of display, PLA2, PLA2+aPLA10, and PLA2+aPLA20 treatments had greater lipid oxidation (P \u3c 0.01) compared to the samples without PLA2. This trend was seen in the other retail display periods. Interestingly, day-7 aPLA10 and aPLA20 had less lipid oxidation than day-7 PL and less oxidation than day-4 PLA2 (P \u3c 0.05). The PL profile analysis showed clear differences between treatments with or without the addition of PLA2. The PLA2 treatments showed greater relative percent of total PL degradation products (P \u3c 0.01) than treatments without PLA2. The PLA2 treatments had less relative percent of total ether-linked phosphatidylcholine (ePC) than treatments without PLA2 (P \u3c 0.01). It appears that aPLA2 had no effect on inhibiting PLA2 hydrolysis as there was no difference (P \u3e 0.10) between PLA2 and aPLA+PLA2 treatments in relative percent of total ePC, phosphatidylcholine (PC), or in PL degradation products. The Bottom Line: Phospholipase-A2 significantly alters beef phospholipids to a composition that is potentially susceptible to lipid oxidation. At day-7 of retail display, there is significant lipid oxidation from PLA2 added treatments, yet the aPLA2 only treatments seem to present an antioxidant effect. Effectively inhibiting PLA2 activity can potentially improve beef shelf-life stability

Host specificity and species colouration mediate the regional decline of nocturnal moths in central European forests

The high diversity of insects has limited the volume of long-term community data with a high taxonomic resolution and considerable geographic replications, especially in forests. Therefore, trends and causes of changes are poorly understood. Here we analyse trends in species richness, abundance and biomass of nocturnal macro moths in three quantitative data sets collected over four decades in forests in southern Germany. Two local data sets, one from coppiced oak forests and one from high oak forests included 125K and 48K specimens from 559 and 532 species, respectively. A third regional data set, representing all forest types in the temperate zone of central Europe comprised 735K specimens from 848 species. Generalized additive mixed models revealed temporal declines in species richness (−38%), abundance (−53%) and biomass (−57%) at the regional scale. These were more pronounced in plant host specialists and in dark coloured species. In contrast, the local coppiced oak forests showed an increase, in species richness (+62%), while the high oak forests showed no clear trends. Left and right censoring as well as cross validation confirmed the robustness of the analyses, which led to four conclusions. First, the decline in insects appears in hyper diverse insect groups in forests and affects species richness, abundance and biomass. Second, the pronounced decline in host specialists suggests habitat loss as an important driver of the observed decline. Third, the more severe decline in dark species might be an indication of global warming as a potential driver. Fourth, the trends in coppiced oak forests indicate that maintaining complex and diverse forest ecosystems through active management may be a promising conservation strategy in order to counteract negative trends in biodiversity, alongside rewilding approaches

The chance of freezing – a conceptional study to parameterize temperature-dependent freezing by including randomness of ice-nucleating particle concentrations

Ice-nucleating particle concentrations (INPCs) can spread over several orders of magnitude at any given temperature. However, this variability is rarely accounted for in heterogeneous ice-nucleation parameterizations. In this paper, we present an approach to incorporate the random variation in the INPC into the parameterization of immersion freezing and analyze this novel concept with various sensitivity tests. In the new scheme, the INPC is drawn from a relative frequency distribution of cumulative INPCs. At each temperature, this distribution describing the INPCs is expressed as a lognormal frequency distribution. The new parameterization scheme does not require aerosol information from the driving model to represent the heterogeneity of INPCs. The scheme's performance is tested in a large-eddy simulation of a relatively warm Arctic mixed-phase stratocumulus. We find that it leads to reasonable ice masses in the cloud, especially when compared to immersion freezing schemes that yield one fixed INPC per temperature and lead to almost no ice production in the simulated cloud. The scheme is sensitive to the median of the frequency distribution and highly sensitive to the standard deviation of the distribution, as well as to the frequency of drawing a new INPC and the resolution of the model. Generally, a higher probability of drawing large INPCs leads to substantially more ice in the simulated cloud. We expose inherent challenges to introducing such a parameterization and explore possible solutions and potential developments.</p

Coal fly ash: Linking immersion freezing behavior and physicochemical particle properties

To date, only a few studies have investigated the potential of coal fly ash particles to trigger heterogeneous ice nucleation in cloud droplets. The presented measurements aim at expanding the sparse dataset and improving process understanding of how physicochemical particle properties can influence the freezing behavior of coal fly ash particles immersed in water. Firstly, immersion freezing measurements were performed with two single particle techniques, i.e., the Leipzig Aerosol Cloud Interaction Simulator (LACIS) and the SPectrometer for Ice Nuclei (SPIN). The effect of suspension time on the efficiency of the coal fly ash particles when immersed in a cloud droplet is analyzed based on the different residence times of the two instruments and employing both dry and wet particle generation. Secondly, two cold-stage setups, one using microliter sized droplets (Leipzig Ice Nucleation Array) and one using nanoliter sized droplets (WeIzmann Supercooled Droplets Observation on Microarray setup) were applied. We found that coal fly ash particles are comparable to mineral dust in their immersion freezing behavior when being dry generated. However, a significant decrease in immersion freezing efficiency was observed during experiments with wet-generated particles in LACIS and SPIN. The efficiency of wet-generated particles is in agreement with the cold-stage measurements. In order to understand the reason behind the deactivation, a series of chemical composition, morphology, and crystallography analyses (single particle mass spectrometry, scanning electron microscopy coupled with energy dispersive X-ray microanalysis, X-ray diffraction analysis) were performed with dry- and wet-generated particles. From these investigations, we conclude that anhydrous CaSO4 and CaO - which, if investigated in pure form, show the same qualitative immersion freezing behavior as observed for dry-generated coal fly ash particles - contribute to triggering heterogeneous ice nucleation at the particle-water interface. The observed deactivation in contact with water is related to changes in the particle surface properties which are potentially caused by hydration of CaSO4 and CaO. The contribution of coal fly ash to the ambient population of ice-nucleating particles therefore depends on whether and for how long particles are immersed in cloud droplets

Challenges in measuring nitrogen isotope signatures in inorganic nitrogen forms: An interlaboratory comparison of three common measurement approaches

Rationale Stable isotope approaches are increasingly applied to better understand the cycling of inorganic nitrogen (Ni) forms, key limiting nutrients in terrestrial and aquatic ecosystems. A systematic comparison of the accuracy and precision of the most commonly used methods to analyze δ15N in NO3− and NH4+ and interlaboratory comparison tests to evaluate the comparability of isotope results between laboratories are, however, still lacking. Methods Here, we conducted an interlaboratory comparison involving 10 European laboratories to compare different methods and laboratory performance to measure δ15N in NO3− and NH4+. The approaches tested were (a) microdiffusion (MD), (b) chemical conversion (CM), which transforms Ni to either N2O (CM-N2O) or N2 (CM-N2), and (c) the denitrifier (DN) methods. Results The study showed that standards in their single forms were reasonably replicated by the different methods and laboratories, with laboratories applying CM-N2O performing superior for both NO3− and NH4+, followed by DN. Laboratories using MD significantly underestimated the “true” values due to incomplete recovery and also those using CM-N2 showed issues with isotope fractionation. Most methods and laboratories underestimated the at%15N of Ni of labeled standards in their single forms, but relative errors were within maximal 6% deviation from the real value and therefore acceptable. The results showed further that MD is strongly biased by nonspecificity. The results of the environmental samples were generally highly variable, with standard deviations (SD) of up to ± 8.4‰ for NO3− and ± 32.9‰ for NH4+; SDs within laboratories were found to be considerably lower (on average 3.1‰). The variability could not be connected to any single factor but next to errors due to blank contamination, isotope normalization, and fractionation, and also matrix effects and analytical errors have to be considered