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Abstract Background The organism-wide effects of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral infection are well studied, but little is known about the dynamics of how the infection spreads in time among or within cells due to the scarcity of suitable high-resolution experimental systems. It has been reported that SARS-CoV-2 infection pathways converge at calcium influx and subcellular calcium distribution changes. Imaging combined with a proper staining technique is an effective tool for studying subcellular calcium-related infection and replication mechanisms at such resolutions. Methods Using two-photon (2P) fluorescence imaging with our novel Ca-selective dye, automated image analysis and clustering analysis were applied to reveal titer and variant effects on SARS-CoV-2-infected Vero E6 cells. Results The application of a new calcium sensor molecule is shown, combined with a high-end 2P technique for imaging and identifying the patterns associated with cellular infection damage within cells. Vero E6 cells infected with SARS-CoV-2 variants, D614G or B.1.1.7, exhibit elevated cytosolic calcium levels, allowing infection monitoring by tracking the cellular changes in calcium level by the internalized calcium sensor. The imaging provides valuable information on how the level and intracellular distribution of calcium are perturbed during the infection. Moreover, two-photon calcium sensing allowed the distinction of infections by two studied viral variants via cluster analysis of the image parameters. This approach will facilitate the study of cellular correlates of infection and their quantification depending on viral variants and viral load. Conclusions We propose a new two-photon microscopy-based method combined with a cell-internalized sensor to quantify the level of SARS-CoV-2 infection. We optimized the applied dye concentrations to not interfere with viral fusion and viral replication events. The presented method ensured the proper monitoring of viral infection, replication, and cell fate. It also enabled distinguishing intracellular details of cell damage, such as vacuole and apoptotic body formation. Using clustering analysis, 2P microscopy calcium fluorescence images were suitable to distinguish two different viral variants in cell cultures. Cellular harm levels read out by calcium imaging were quantitatively related to the initial viral multiplicity of infection numbers. Thus, 2P quantitative calcium imaging might be used as a correlate of infection or a correlate of activity in cellular antiviral studies. Graphical Abstract

Summary: Human single-stranded DNA binding protein 1 (hSSB1/NABP2/OBFC2B) plays central roles in DNA repair. Here, we show that purified hSSB1 undergoes redox-dependent liquid-liquid phase separation (LLPS) in the presence of single-stranded DNA or RNA, features that are distinct from those of LLPS by bacterial SSB. hSSB1 nucleoprotein droplets form under physiological ionic conditions in response to treatment modeling cellular oxidative stress. hSSB1’s intrinsically disordered region is indispensable for LLPS, whereas all three cysteine residues of the oligonucleotide/oligosaccharide-binding fold are necessary to maintain redox-sensitive droplet formation. Proteins interacting with hSSB1 show selective enrichment inside hSSB1 droplets, suggesting tight content control and recruitment functions for the condensates. While these features appear instrumental for genome repair, we detected cytoplasmic hSSB1 condensates in various cell lines colocalizing with stress granules upon oxidative stress, implying extranuclear function in cellular stress response. Our results suggest condensation-linked roles for hSSB1, linking genome repair and cytoplasmic defense.

This research assesses the effects of drought and salt stresses applied individually or in combination on Thymus pannonicus, a resilient perennial herb native to the Pannonian vegetation region. Our experiment aimed to comprehensively evaluate morphological, physiological, and biochemical parameters of the thyme subjected to two levels of soil water capacity (SWC) - 70 % (control and salt stress) and 40 % (drought stress and combined stresses) - along with two salinity levels, 0 mM NaCl (control and drought stress) and 60 mM NaCl (salt stress and combined stresses). The plants exhibited significant responses to all the studied parameters under 40 % SWC in both treatments, demonstrating notable reductions in shoot and root parameters. In contrast, salt treatment at 60 mM had a minimal impact on growth. A stability in photosynthetic pigment composition was observed in all treatment, whereas a notable decrease (p < 0.05) in relative water content was seen in drought-stressed treatments compared to the to the control. Essential oil contents were higher in the control (0.91 ml/100g DW) but significantly declined (p < 0.05) under drought stress and combined stresses conditions (0.49 ml/100g DW and 0.53 ml/100g DW respectively). Particularly, the GC-MS analysis revealed a significant decrease (p < 0.05) in thymol and carvacrol content for the drought-stressed treatments. Substantial reductions were evident for the total polyphenol concentration and antioxidant capacity under the combined stress treatment, measuring 101.49 mg GAE/g DW and 106.57 AAE/g DW, respectively. Interestingly, the HPLC analysis indicated the highest levels of rosmarinic acid and caffeic acid in the drought-stressed treatment, while the lowest ones were recorded in the combined stress treatment. These findings indicated the responses of Thymus pannonicus to environmental stressors, emphasizing the need for further investigations for a comprehensive understanding of salinity effects.

While fungus-resistant grape varieties (FRGVs) offer comprehensive sustainable benefits, their slow adoption and consumer preferences warrant investigation. Therefore, a discrete choice model for key parameters influencing consumer perceptions, including product attributes, pricing, grape variety preferences, labelling and information dissemination, was constructed based on a representative sample of 1276 wine consumers in Germany. The data were analysed using a conditional logit utility model to gain insights into the preferences and willingness to pay (WTP) of each product attribute. Due to the low level of awareness of these grape varieties, the sample was divided into two groups: a control group and an experimental group, which was conditioned prior to the study to investigate the market potential of FRGVs. The results show that the conditioned group exhibited a stronger inclination towards FRGVs, indicating an increased awareness of environmentally friendly practices. Price sensitivity was influenced by conditioning, with consumers being more sensitive to higher prices after being informed. The study highlights the importance of informed consumer education in promoting environmentally sustainable products like wine from FRGVs.

Abstract The Eurasian woodcock prefers habitats where its main prey, earthworms, can be found in higher densities. Although they are forest‐dwelling birds, they regularly visit pastures and natural grasslands at night, where earthworm abundance is generally higher. However, there is little information on fine‐scale habitat use in relation to variation in habitat characteristics and prey availability, particularly beyond the breeding season. In our study, we investigated if the nocturnal occurrence of woodcocks during migratory stopover periods differed between two neighbouring fields, or management units, with similar vegetation structure, and if within‐field variation in the spatial patterns of woodcock sightings were associated with fine‐scale earthworm densities and soil parameters. Specifically, we used GPS tracking data of two tagged woodcocks and direct observation data to study patterns of occurrence of birds in a mixed forest‐pasture landscape in Hungary during pre‐ and post‐breeding periods. We compared these patterns with fine‐scale soil characteristics and earthworm abundance, acquired by field sampling. We found that the field with higher earthworm abundance was visited by woodcocks more frequently, and this correlation was similarly observed at the intra‐field level. Our results demonstrate that woodcocks select foraging sites with higher earthworm densities at multiple spatial scales, both between fields (coarse scale), and within fields (fine‐scale). Considering that woodcocks tended to return to the same field to forage at night, the strong associations between occupancy and resources provide a basis for developing habitat management strategies at the field level for conservation. As earthworm densities and soil parameters are good indicators of woodcock foraging habitat, measuring those variables, at least at a coarse scale, could aid in predicting important habitats for the species across the landscape.

This paper presents the approach adopted by the EGI-ACE project for the setup and delivery of Data Spaces for various scientific domains. The work was implemented by members of the EGI e-infrastructure and of several European Research Infrastructures in the context of the European Open Science Cloud programme. Our results are several Data Space services that enable the reuse and exploitation of open, scientific big data for compute intensive use cases. The paper illustrates the EGI-ACE approach through two examples: (1) EMSO ERIC Data Portal for seafloor and water column research and (2) ENES Data Space for climate research.

Summary: Retinal ganglion cells (RGCs) summate inputs and forward a spike train code to the brain in the form of either maintained spiking (sustained) or a quickly decaying brief spike burst (transient). We report diverse response transience values across the RGC population and, contrary to the conventional transient/sustained scheme, responses with intermediary characteristics are the most abundant. Pharmacological tests showed that besides GABAergic inhibition, gap junction (GJ)–mediated excitation also plays a pivotal role in shaping response transience and thus visual coding. More precisely GJs connecting RGCs to nearby amacrine and RGCs play a defining role in the process. These GJs equalize kinetic features, including the response transience of transient OFF alpha (tOFFα) RGCs across a coupled array. We propose that GJs in other coupled neuron ensembles in the brain are also critical in the harmonization of response kinetics to enhance the population code and suit a corresponding task.

Abstract Glyphosate (GLY), the most widely used herbicide active ingredient (AI) in the world, is frequently detected in aquatic environments where it can affect non-target organisms. Globally, more than 2000 commercial GLY-based herbicides (GBHs) are used to control weeds. Non-target organisms are exposed to complex pesticide formulations under real environmental conditions, but the co-formulants contained in GBHs are classified as so-called inert and inactive ingredients in terms of their biological effects. The main objective of this comprehensive review is to compile the results of aquatic ecotoxicological studies on the side-effects of GLY, GBHs, and their formulating agents. Based on the results demonstrated for a variety of plant and animal aquatic organisms, oxidative stress appears to be a major trigger for these adverse effects, affecting the integrity of DNA and other biochemical functions. Furthermore, there is evidence of impairment of various physiological and behavioral functions. Adverse effects of GLY and GBHs have been observed even at very low concentrations. There are also differences in the sensitivity of the aquatic organisms tested, even with similar lifestyles, habitats or identical taxa. The studies typically investigate the short-term effects of a single exposure to GLY/GBH on a single species, whilst in reality multiple applications of GBHs together with other pesticides are common during a cropping cycle. Moreover, the interactions between GLY/GBHs and other aquatic contaminants are rarely studied. Higher toxicity of GBHs compared to GLY alone has often been observed, demonstrating that co-formulants can be highly toxic on their own and markedly increase the toxicity of the GBH formulation. The possible impurities in GBHs, such as heavy metals, can cause additional problems for the environment and food safety. The widespread and massive use of GBHs leads to increased exposure and environmental hazards. In addition, the need for a revision of the risk assessment system is emphasized. According to the results of aquatic ecotoxicological studies, the current use and pollution of the aquatic environment by GLY/GBHs is highly problematic and cannot be considered environmentally sustainable. It is, therefore, necessary to at least tighten the permitted forms of use.

Sustainable wildlife management requires reliable data on population, habitat, and the interaction between them. Since 1993, Hungary has maintained the National Game Management Database, which collects data on game populations and hunting bags annually. During the last three decades, populations of medium-sized predators have remarkably increased in the country. Our study analyzed changes in hunting bag data and spatial distribution of two invasive alien species, the raccoon (Procyon lotor) and the raccoon dog (Nyctereutes procyonoides), utilizing annual game management reports from Game Management Units from 1997 to 2021. For spatial analysis, we employed a UTM grid system (10 × 10 km) covering the country. Our findings confirmed increasing trends in the hunting bag and area (number of grid cells) containing hunting bags for both species. From the first (1997–2001) to the last (2017–2021) period, the average annual hunting bag increased (raccoon: 0.40 → 11.00 ind/year; raccoon dog: 3.40 → 8.00 ind/year), and the average number of cells with hunting bags also expanded significantly (raccoon: 2.00 → 9.60 UTM cells/year; raccoon dog: 5.80 → 15.20 UTM cells/year). These trends and the size of potentially suitable habitats suggest a further spread and population increase, posing challenges for nature conservationists and wildlife managers.

To understand the processes behind pollinator declines and for the conservation of pollination services, we need to understand fundamental drivers influencing pollinator behaviour. Here, we aimed to elucidate how wild bumblebees interact with three plant species and investigated their foraging behaviour with varying flower densities. We video-recorded Bombus terrestris in 60 × 60 cm quadrats of Lotus creticus, Persicaria capitata, and Trifolium pratense in urban areas of Terceira (Azores, Portugal). For the automated bumblebee detection and counting, we created deep learning-based computer vision models with custom datasets. We achieved high model accuracy of 0.88 for Lotus and Persicaria and 0.95 for Trifolium, indicating accurate bumblebee detection. In our study, flower cover was the only factor that influenced the attractiveness of flower patches, and plant species did not have an effect. We detected a significant positive effect of flower cover on the attractiveness of flower patches for flower-visiting bumblebees. The time spent per unit of inflorescence surface area was longer on the Trifolium than those on the Lotus and Persicaria. However, our result did not indicate significant differences in the time bumblebees spent on inflorescences among the three plant species. Here, we also justify computer vision-based analysis as a reliable tool for studying pollinator behavioural ecology.

The study presents the evaluation of a radiocarbon series, currently unparalleled in the research of the early medieval Carpathian Basin, which comprises data from the 7th to the 10th century AD. We provide a data set that, when combined with the radiocarbon data available in the related literature, covers the period in focus. The results of its analysis can be considered novel in several respects: 1) the radiocarbon data sequence and the relative chronological framework established for the Late Avar Period concord, 2) based on the radiocarbon sequence, the Middle Avar Period in certain large cemeteries (i.e., Tiszafüred-Majoros) started considerably earlier than it was assumed previously, based on ‘Middle Avar Period’ elite graves—and, interestingly, earlier even than the coin-dated ‘Middle Avar’ elite grave horizon, and 3) the data of the latest grave horizon in Avar cemeteries suggests a similar asynchronism between the related sites. The data set allows one to draw preliminary conclusions about the trends of the early medieval cultural and social transformations in the Carpathian Basin and outline ‘innovative’ groups which, by maintaining contacts with diverse regions outside the Carpathian Basin, played a central role in these processes.

Abstract The BCL-2 inhibitor Venetoclax is a promising agent for the treatment of acute myeloid leukemia (AML). However, many patients are refractory to Venetoclax, and resistance develops quickly. ATP-binding cassette (ABC) transporters mediate chemotherapy resistance but their role in modulating the activity of targeted small-molecule inhibitors is unclear. Using CRISPR/Cas9 screening, we find that loss of ABCC1 strongly increases the sensitivity of AML cells to Venetoclax. Genetic and pharmacologic ABCC1 inactivation potentiates the anti-leukemic effects of BCL-2 inhibitors and efficiently re-sensitizes Venetoclax-resistant leukemia cells. Conversely, ABCC1 overexpression induces resistance to BCL-2 inhibitors by reducing intracellular drug levels, and high ABCC1 levels predicts poor response to Venetoclax therapy in patients. Consistent with ABCC1-specific export of glutathionylated substrates, inhibition of glutathione metabolism increases the potency of BCL-2 inhibitors. These results identify ABCC1 and glutathione metabolism as mechanisms limiting efficacy of BCL-2 inhibitors, which may pave the way to development of more effective therapies.