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Incorporation of noncanonical amino acids (ncAAs) with bioorthogonal reactive groups by amber suppression allows the generation of synthetic proteins with desired novel properties. Such modified molecules are in high demand for basic research and therapeutic applications such as cancer treatment and in vivo imaging. The positioning of the ncAA-responsive codon within the protein's coding sequence is critical in order to maintain protein function, achieve high yields of ncAA-containing protein, and allow effective conjugation. Cell-free ncAA incorporation is of particular interest due to the open nature of cell-free systems and their concurrent ease of manipulation. In this study, we report a straightforward workflow to inquire ncAA positions in regard to incorporation efficiency and protein functionality in a Chinese hamster ovary (CHO) cell-free system. As a model, the well-established orthogonal translation components Escherichia coli tyrosyl-tRNA synthetase (TyrRS) and tRNATyr(CUA) were used to site-specifically incorporate the ncAA p-azido-l-phenylalanine (AzF) in response to UAG codons. A total of seven ncAA sites within an anti-epidermal growth factor receptor (EGFR) single-chain variable fragment (scFv) N-terminally fused to the red fluorescent protein mRFP1 and C-terminally fused to the green fluorescent protein sfGFP were investigated for ncAA incorporation efficiency and impact on antigen binding. The characterized cell-free dual fluorescence reporter system allows screening for ncAA incorporation sites with high incorporation efficiency that maintain protein activity. It is parallelizable, scalable, and easy to operate. We propose that the established CHO-based cell-free dual fluorescence reporter system can be of particular interest for the development of antibody-drug conjugates (ADCs).
We study the first-arrival (first-hitting) dynamics and efficiency of a one-dimensional random search model performing asymmetric Levy flights by leveraging the Fokker-Planck equation with a delta-sink and an asymmetric space-fractional derivative operator with stable index alpha and asymmetry (skewness) parameter beta.
We find exact analytical results for the probability density of first-arrival times and the search efficiency, and we analyse their behaviour within the limits of short and long times.
We find that when the starting point of the searcher is to the right of the target, random search by Brownian motion is more efficient than Levy flights with beta <= 0 (with a rightward bias) for short initial distances, while for beta>0 (with a leftward bias) Levy flights with alpha -> 1 are more efficient.
When increasing the initial distance of the searcher to the target, Levy flight search (except for alpha=1 with beta=0) is more efficient than the Brownian search. Moreover, the asymmetry in jumps leads to essentially higher efficiency of the Levy search compared to symmetric Levy flights at both short and long distances, and the effect is more pronounced for stable indices alpha close to unity.
Soil bacteria play a fundamental role in pedogenesis. However, knowledge about both the impact of climate and slope aspects on microbial communities and the consequences of these items in pedogenesis is lacking. Therefore, soil-bacterial communities from four sites and two different aspects along the climate gradient of the Chilean Coastal Cordillera were investigated. Using a combination of microbiological and physicochemical methods, soils that developed in arid, semi-arid, mediterranean, and humid climates were analyzed. Proteobacteria, Acidobacteria, Chloroflexi, Verrucomicrobia, and Planctomycetes were found to increase in abundance from arid to humid climates, while Actinobacteria and Gemmatimonadetes decreased along the transect. Bacterial-community structure varied with climate and aspect and was influenced by pH, bulk density, plant-available phosphorus, clay, and total organic-matter content. Higher bacterial specialization was found in arid and humid climates and on the south-facing slope and was likely promoted by stable microclimatic conditions. The presence of specialists was associated with ecosystem-functional traits, which shifted from pioneers that accumulated organic matter in arid climates to organic decomposers in humid climates. These findings provide new perspectives on how climate and slope aspects influence the composition and functional capabilities of bacteria, with most of these capabilities being involved in pedogenetic processes.
Genomic prediction has revolutionized crop breeding despite remaining issues of transferability of models to unseen environmental conditions and environments. Usage of endophenotypes rather than genomic markers leads to the possibility of building phenomic prediction models that can account, in part, for this challenge. Here, we compare and contrast genomic prediction and phenomic prediction models for 3 growth-related traits, namely, leaf count, tree height, and trunk diameter, from 2 coffee 3-way hybrid populations exposed to a series of treatment-inducing environmental conditions. The models are based on 7 different statistical methods built with genomic markers and ChlF data used as predictors. This comparative analysis demonstrates that the best-performing phenomic prediction models show higher predictability than the best genomic prediction models for the considered traits and environments in the vast majority of comparisons within 3-way hybrid populations. In addition, we show that phenomic prediction models are transferrable between conditions but to a lower extent between populations and we conclude that chlorophyll a fluorescence data can serve as alternative predictors in statistical models of coffee hybrid performance. Future directions will explore their combination with other endophenotypes to further improve the prediction of growth-related traits for crops.
Reducing greenhouse gas emissions in food systems is becoming more challenging as food is increasingly consumed away from producer regions, highlighting the need to consider emissions embodied in trade in agricultural emissions accounting.
To address this, our study explores recent trends in trade-adjusted agricultural emissions of food items at the global, regional, and national levels.
We find that emissions are largely dependent on a country’s consumption patterns and their agricultural emission intensities relative to their trading partners’.
The absolute differences between the production-based and trade-adjusted emissions accounting approaches are especially apparent for major agricultural exporters and importers and where large shares of emission-intensive items such as ruminant meat, milk products and rice are involved.
In relative terms, some low-income and emerging and developing economies with consumption of high emission intensity food products show large differences between approaches.
Similar trends are also found under various specifications that account for trade and re-exports differently.
These findings could serve as an important element towards constructing national emissions reduction targets that consider trading partners, leading to more effective emissions reductions overall.
The Lena Delta in Siberia is the largest delta in the Arctic and as a snow-dominated ecosystem particularly vulnerable to climate change.
Using the two decades of MODerate resolution Imaging Spectroradiometer satellite acquisitions, this study investigates interannual and spatial variability of snow-cover duration and summer vegetation vitality in the Lena Delta.
We approximated snow by the application of the normalized difference snow index and vegetation greenness by the normalized difference vegetation index (NDVI). We consolidated the analyses by integrating reanalysis products on air temperature from 2001 to 2021, and air temperature, ground temperature, and the date of snow-melt from time-lapse camera (TLC) observations from the Samoylov observatory located in the central delta.
We extracted spring snow-cover duration determined by a latitudinal gradient. The 'regular year' snow-melt is transgressing from mid-May to late May within a time window of 10 days across the delta.
We calculated yearly deviations per grid cell for two defined regions, one for the delta, and one focusing on the central delta. We identified an ensemble of early snow-melt years from 2012 to 2014, with snow-melt already starting in early May, and two late snow-melt years in 2004 and 2017, with snow-melt starting in June. In the times of TLC recording, the years of early and late snow-melt were confirmed.
In the three summers after early snow-melt, summer vegetation greenness showed neither positive nor negative deviations. Whereas, vegetation greenness was reduced in 2004 after late snow-melt together with the lowest June monthly air temperature of the time series record. Since 2005, vegetation greenness is rising, with maxima in 2018 and 2021.
The NDVI rise since 2018 is preceded by up to 4 degrees C warmer than average June air temperature. The ongoing operation of satellite missions allows to monitor a wide range of land surface properties and processes that will provide urgently needed data in times when logistical challenges lead to data gaps in land-based observations in the rapidly changing Arctic.
The concepts of CO2 emission, global warming, climate change, and their environmental impacts are of utmost importance for the understanding and protection of the ecosystems.
Among the natural sources of gases into the atmosphere, the contribution of geogenic sources plays a crucial role. However, while subaerial emissions are widely studied, submarine outgassing is not yet well understood.
In this study, we review and catalog 122 literature and unpublished data of submarine emissions distributed in ten coastal areas of the Aegean Sea. This catalog includes descriptions of the degassing vents through in situ observations, their chemical and isotopic compositions, and flux estimations.
Temperatures and pH data of surface seawaters in four areas affected by submarine degassing are also presented.
This overview provides useful information to researchers studying the impact of enhanced seawater CO2 concentrations related either to increasing CO2 levels in the atmosphere or leaking carbon capture and storage systems.
The production of volatile organic compounds (VOCs) represents a promising strategy of plant-beneficial bacteria to control soil-borne phytopathogens.
Pseudomonas sp. PICF6 and Pseudomonas simiae PICF7 are two indigenous inhabitants of olive roots displaying effective biological control against Verticillium dahliae. Additionally, strain PICF7 is able to promote the growth of barley and Arabidopsis thaliana, VOCs being involved in the growth of the latter species.
In this study, the antagonistic capacity of these endophytic bacteria against relevant phytopathogens (Verticillium spp., Rhizoctonia solani, Sclerotinia sclerotiorum and Fusarium oxysporum f.sp. lycopersici) was assessed. Under in vitro conditions, PICF6 and PICF7 were only able to antagonize representative isolates of V. dahliae and V. longisporum. Remarkably, both strains produced an impressive portfolio of up to twenty VOCs, that included compounds with reported antifungal (e.g., 1-undecene, (methyldisulfanyl) methane and 1-decene) or plant growth promoting (e.g., tridecane, 1-decene) activities. Moreover, their volatilomes differed strongly in the absence and presence of V. dahliae.
For example, when co incubated with the defoliating pathotype of V. dahliae, the antifungal compound 4-methyl-2,6-bis(2-methyl-2-propanyl)phenol was produced. Results suggest that volatiles emitted by these endophytes may differ in their modes of action, and that potential benefits for the host needs further investigation in planta.
How tone, intonation and emotion shape the development of infants' fundamental frequency perception
(2022)
Fundamental frequency (integral(0)), perceived as pitch, is the first and arguably most salient auditory component humans are exposed to since the beginning of life.
It carries multiple linguistic (e.g., word meaning) and paralinguistic (e.g., speakers' emotion) functions in speech and communication.
The mappings between these functions and integral(0) features vary within a language and differ cross-linguistically. For instance, a rising pitch can be perceived as a question in English but a lexical tone in Mandarin. Such variations mean that infants must learn the specific mappings based on their respective linguistic and social environments.
To date, canonical theoretical frameworks and most empirical studies do not view or consider the multi-functionality of integral(0), but typically focus on individual functions. More importantly, despite the eventual mastery of integral(0) in communication, it is unclear how infants learn to decompose and recognize these overlapping functions carried by integral(0). In this paper, we review the symbioses and synergies of the lexical, intonational, and emotional functions that can be carried by integral(0) and are being acquired throughout infancy.
On the basis of our review, we put forward the Learnability Hypothesis that infants decompose and acquire multiple integral(0) functions through native/environmental experiences. Under this hypothesis, we propose representative cases such as the synergy scenario, where infants use visual cues to disambiguate and decompose the different integral(0) functions. Further, viable ways to test the scenarios derived from this hypothesis are suggested across auditory and visual modalities.
Discovering how infants learn to master the diverse functions carried by integral(0) can increase our understanding of linguistic systems, auditory processing and communication functions.
In-depth understanding of the reorganization of the hydrological cycle in response to global climate change is crucial in highly sensitive regions like the eastern Mediterranean, where water availability is a major factor for socioeconomic and political development.
The sediments of Lake Lisan provide a unique record of hydroclimatic change during the last glacial to Holocene transition (ca. 24-11 ka) with its tremendous water level drop of similar to 240 m that finally led to its transition into the present hypersaline water body-the Dead Sea.
Here we utilize high-resolution sedimentological analyses from the marginal terraces and deep lake to reconstruct an unprecedented seasonal record of the last millennia of Lake Lisan. Aragonite varve formation in intercalated intervals of our record demonstrates that a stepwise long-term lake level decline was interrupted by almost one millennium of rising or stable water level.
Even periods of pronounced water level drops indicated by gypsum deposition were interrupted by decades of positive water budgets.
Our results thus highlight that even during major climate change at the end of the last glacial, decadal to millennial periods of relatively stable or positive moisture supply occurred which could have been an important premise for human sedentism.
The Arctic is rich in aquatic systems and experiences rapid warming due to climate change. The accelerated warming causes permafrost thaw and the mobilization of organic carbon.
When dissolved organic carbon is mobilized, this DOC can be transported to aquatic systems and degraded in the water bodies and further downstream. Here, we analyze the influence of different landscape components on DOC concentrations and export in a small (6.45 km(2)) stream catchment in the Lena River Delta.
The catchment includes lakes and ponds, with the flow path from Pleistocene yedoma deposits across Holocene non-yedoma deposits to the river outlet. In addition to DOC concentrations, we use radiocarbon dating of DOC as well as stable oxygen and hydrogen isotopes (delta O-18 and delta D) to assess the origin of DOC.
We find significantly higher DOC concentrations in the Pleistocene yedoma area of the catchment compared to the Holocene non-yedoma area with medians of 5 and 4.5 mg L-1 (p < 0.05), respectively. When yedoma thaw streams with high DOC concentration reach a large yedoma thermokarst lake, we observe an abrupt decrease in DOC concentration, which we attribute to dilution and lake processes such as mineralization. The DOC ages in the large thermokarst lake (between 3,428 and 3,637 C-14 y BP) can be attributed to a mixing of mobilized old yedoma and Holocene carbon.
Further downstream after the large thermokarst lake, we find progressively younger DOC ages in the stream water to its mouth, paired with decreasing DOC concentrations. This process could result from dilution with leaching water from Holocene deposits and/or emission of ancient yedoma carbon to the atmosphere. Our study shows that thermokarst lakes and ponds may act as DOC filters, predominantly by diluting incoming waters of higher DOC concentrations or by re-mineralizing DOC to CO2 and CH4.
Nevertheless, our results also confirm that the small catchment still contributes DOC on the order of 1.2 kg km(-2) per day from a permafrost landscape with ice-rich yedoma deposits to the Lena River.
Consumers are increasingly demanding higher quality and safety standards for the products they consume, and one of this is wheat flour, the basis of a wide variety of processed products. This major component in the diet of many communities can be contaminated by microorganisms before the grain harvest, or during the grain storage right before processing. These microorganisms include several fungal species, many of which produce mycotoxins, secondary metabolites that can cause severe acute and chronic disorders. Yet, we still know little about the overall composition of fungal communities associated with wheat flour. In this study, we contribute to fill this gap by characterizing the fungal microbiome of different types of wheat flour using culture-dependent and -independent techniques. Qualitatively, these approaches suggested similar results, highlighting the presence of several fungal taxa able to produce mycotoxins. In-vitro isolation of fungal species suggest a higher frequency of Penicillium, while metabarcoding suggest a higher abundance of Alternaria. This discrepancy might reside on the targeted portion of the community (alive vs. overall) or in the specific features of each technique. Thus, this study shows that commercial wheat flour hosts a wide fungal diversity with several taxa potentially representing concerns for consumers, aspects that need more attention throughout the food production chain.
In Decolonizing Universalism: A Transnational Feminist Ethic zielt Serene Khader auf eine Neuausrichtung der feministischen Perspektive, welche es schafft, dekolonial und anti-imperialistisch zu sein, ohne gleichzeitig dem Universalismus komplett abzuschwören. Die Motivation hinter dieser Neuorientierung ist die Einsicht, dass der liberale moralische Universalismus oftmals kulturelle Vorherrschaft und Imperialismus verstärkt. In diesem Kommentar wollen wir (a) uns mit der Frage beschäftigen, was genau unter Geschlechtergerechtigkeit verstanden werden soll und welcher Maßstab zur Beantwortung der Frage nach Gerechtigkeit angebracht ist und (b) einige Ideen zum Unterschied zwischen idealer und nicht-idealer Theorie liefern.
Die Debatte um epistemische Ungerechtigkeit verbindet normative Gerechtigkeitstheorien mit erkenntnistheoretischen Theorien und stellt somit die Art von wichtigen Fragen, die in den letzten Jahren sowohl innerhalb als auch außerhalb der Wissenschaft internationale Aufmerksamkeit erfahren haben. Verwiesen sei hier etwa auf soziale Bewegungen wie #MeToo und #BlackLivesMatter zeigen. Theorien der epistemischen Ungerechtigkeit (sowie verwandte Theorien wie Epistemologie des Unwissens, feministische Erkenntnistheorie und Standpunkttheorie) können sowohl epistemische Praktiken analysieren und einen Beitrag zu Gerechtigkeitstheorien und sozialer Epistemologie liefern, als auch zu adäquateren Verständnissen von existierenden Ungerechtigkeiten beitragen. In dem hier vorliegenden Schwerpunkt werden Beiträge zu eben solchen bislang wenig erforschten Ungerechtigkeiten sowie neue Diskussionsbeiträge zur Debatte um epistemische Ungerechtigkeiten geliefert.
Assessing the impact of hydrogen absorption on the characteristics of the Galactic center excess
(2022)
We present a new reconstruction of the distribution of atomic hydrogen in the inner Galaxy that is based on explicit radiation transport modeling of line and continuum emission and a gas-flow model in the barred Galaxy that provides distance resolution for lines of sight toward the Galactic center.
The main benefits of the new gas model are (a) the ability to reproduce the negative line signals seen with the HI4PI survey and (b) the accounting for gas that primarily manifests itself through absorption.
We apply the new model of Galactic atomic hydrogen to an analysis of the diffuse gamma-ray emission from the inner Galaxy, for which an excess at a few GeV was reported that may be related to dark matter.
We find with high significance an improved fit to the diffuse gamma-ray emission observed with the Fermi-LAT, if our new H i model is used to estimate the cosmic-ray induced diffuse gamma-ray emission.
The fit still requires a nuclear bulge at high significance. Once this is included there is no evidence of a dark-matter signal, be it cuspy or cored. But an additional so-called boxy bulge is still favored by the data.
This finding is robust under the variation of various parameters, for example, the excitation temperature of atomic hydrogen, and a number of tests for systematic issues.
The Brassica napus seed microbiota is cultivar-specific and transmitted via paternal breeding lines
(2022)
Seed microbiota influence germination and plant health and have the potential to improve crop performance, but the factors that determine their structure and functions are still not fully understood.
Here, we analysed the impact of plant-related and external factors on seed endophyte communities of 10 different oilseed rape (Brassica napus L.) cultivars from 26 field sites across Europe.
All seed lots harboured a high abundance and diversity of endophytes, which were dominated by six genera: Ralstonia, Serratia, Enterobacter, Pseudomonas, Pantoea, and Sphingomonas.
The cultivar was the main factor explaining the variations in bacterial diversity, abundance and composition. In addition, the latter was significantly influenced by diverse biotic and abiotic factors, for example host germination rates and disease resistance against Plasmodiophora brassicae.
A set of bacterial biomarkers was identified to discriminate between characteristics of the seeds, for example Sphingomonas for improved germination and Brevundimonas for disease resistance.
Application of a Bayesian community approach suggested vertical transmission of seed endophytes, where the paternal parent plays a major role and might even determine the germination performance of the offspring.
This study contributes to the understanding of seed microbiome assembly and underlines the potential of the microbiome to be implemented in crop breeding and biocontrol programmes.
Glucosinolates are plant secondary metabolites found in cruciferous vegetables (Brassicaceae) that are valued for their potential health benefits. Frequently consumed representatives of these vegetables, for example, are white or red cabbage, which are typically boiled before consumption. Recently, 3-alk(en)yl-4-hydroxythiazolidine-2-thiones were identified as a class of thermal glucosinolate degradation products that are formed during the boiling of cabbage. Since these newly discovered compounds are frequently consumed, this raises questions about their potential uptake and their possible bioactive functions. Therefore, 3-allyl-4-hydroxythiazolidine-2-thione (allyl HTT) and 4-hydroxy-3-(4-(methylsulfinyl) butyl)thiazolidine-2-thione (4-MSOB HTT) as degradation products of the respective glucosinolates sinigrin and glucoraphanin were investigated. After consumption of boiled red cabbage broth, recoveries of consumed amounts of the degradation products in urine collected for 24 h were 18 +/- 5% for allyl HTT and 21 +/- 4% for 4-MSOB HTT (mean +/- SD, n = 3). To investigate the stability of the degradation products during uptake and to elucidate the uptake mechanism, both an in vitro stomach and an in vitro intestinal model were applied. The results indicate that the uptake of allyl HTT and 4-MSOB HTT occurs by passive diffusion. Both compounds show no acute cell toxicity, no antioxidant potential, and no change in NAD(P)H dehydrogenase quinone 1 (NQO1) activity up to 100 mu M. However, inhibition of glycogen synthase kinases-3 (GSK-3) in the range of 20% for allyl HTT for the isoform GSK-3 beta and 29% for 4-MSOB HTT for the isoform GSK-3 alpha at a concentration of 100 mu M was found. Neither health-promoting nor toxic effects of 3-alk(en)yl-4-hydroxythiazolidine-2-thiones were found in the four tested assays carried out in this study, which contrasts with the properties of other glucosinolate degradation products, such as isothiocyanates.
The release of greenhouse gases from the large organic carbon stock in permafrost deposits in the circumarctic regions may accelerate global warming upon thaw. The extent of this positive climate feedback is thought to be largely controlled by the microbial degradability of the organic matter preserved in these sediments. In addition, weathering and oxidation processes may release inorganic carbon preserved in permafrost sediments as CO2, which is generally not accounted for. We used C-13 and C-14 analysis and isotopic mass balances to differentiate and quantify organic and inorganic carbon released as CO2 in the field from an active retrogressive thaw slump of Pleistocene-age Yedoma and during a 1.5-years incubation experiment. The results reveal that the dominant source of the CO2 released from freshly thawed Yedoma exposed as thaw mound is Pleistocene-age organic matter (48-80%) and to a lesser extent modern organic substrate (3-34%). A significant portion of the CO2 originated from inorganic carbon in the Yedoma (17-26%). The mixing of young, active layer material with Yedoma at a site on the slump floor led to the preferential mineralization of this young organic carbon source. Admixtures of younger organic substrates in the Yedoma thaw mound were small and thus rapidly consumed as shown by lower contributions to the CO2 produced during few weeks of aerobic incubation at 4 degrees C corresponding to approximately one thaw season. Future CO2 fluxes from the freshly thawed Yedoma will contain higher proportions of ancient inorganic (22%) and organic carbon (61-78%) as suggested by the results at the end, after 1.5 years of incubation. The increasing contribution of inorganic carbon during the incubation is favored by the accumulation of organic acids from microbial organic matter degradation resulting in lower pH values and, in consequence, in inorganic carbon dissolution. Because part of the inorganic carbon pool is assumed to be of pedogenic origin, these emissions would ultimately not alter carbon budgets. The results of this study highlight the preferential degradation of younger organic substrates in freshly thawed Yedoma, if available, and a substantial release of CO2 from inorganic sources.
Urokinase-type plasminogen activator is widely discussed as a marker for cancer prognosis and diagnosis and as a target for cancer therapies. Together with its receptor, uPA plays an important role in tumorigenesis, tumor progression and metastasis. In the present study, systematic evolution of ligands by exponential enrichment (SELEX) was used to select single-stranded DNA aptamers targeting different forms of human uPA. Selected aptamers allowed the distinction between HMW-uPA and LMW-uPA, and therefore, presumably, have different binding regions. Here, uPAapt-02-FR showed highly affine binding with a K-D of 0.7 nM for HMW-uPA and 21 nM for LMW-uPA and was also able to bind to pro-uPA with a K-D of 14 nM. Furthermore, no cross-reactivity to mouse uPA or tissue-type plasminogen activator (tPA) was measured, demonstrating high specificity. Suppression of the catalytic activity of uPA and inhibition of uPAR-binding could be demonstrated through binding with different aptamers and several of their truncated variants. Since RNA aptamers are already known to inhibit uPA-uPAR binding and other pathological functions of the uPA system, these aptamers represent a novel, promising tool not only for detection of uPA but also for interfering with the pathological functions of the uPA system by additionally inhibiting uPA activity.